Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.006
P. Lei, Chaofeng Tang, W. Bu, Songning Yu
Objective To investigate the relationship between solute carrier transporter 22A3 (SLC22A3) gene rs7758229 polymorphism and prognosis of pancreatic cancer. Methods One hundred patients with resectable pancreatic cancer who underwent surgery were admitted to our hospital from March 30, 2014 to March 30, 2016 were selected. There were were 61 male and 39 female patients, aging from 35 to 79 years with a median age of (49.1±5.7) years. The location included head and neck of the pancreas (n=66), body and tail of pancreas (n=34). According to the 7th American Joint Committee on Cancer (AJCC) Pancreatic Cancer Staging System, 19 cases were in stage I, 56 cases were in stage Ⅱ and 25 cases in stage Ⅲ. On the 2nd day after admission, 5 ml of fasting venous blood was taken, and the polymorphism of SLC22A3 gene rs7758229 was detected by DNA extraction kit and sequencing method. The patients were followed up and the end point was all-cause death. The correlation between polymorphism of rs7758229 and the prognosis of pancreatic cancer, and the factors affecting the overall survival (OS) of the patients were analyzed. The date analysis was conducted by SPSS 25.0; the gene type distribution deviation was tested by Hardy-Weinberg equilibrium, the correlation analysis of clinical date was analyzed by χ2 test analysis. The correlation between polymorphism of rs7758229 and the prognosis of pancreatic cancer, and the factors affecting the OS of the patients were analyzed by by Kaplan- Meier method and Cox model respectively. Results There were 28 cases (28.0%) of GG type, 51 cases (51.0%) of GT type, and 21 cases (21.0%) of TT type. The rs7758229 mononucleic acid polymorphism was significantly correlated with the degree of tumor differentiation and clinical stage (χ2=10.209, 10.826, P<0.05). Six patients were lost to follow-up (2 patients with GG, 3 patients with GT, and 1 patient with TT) with a median follow-up of 46 months. There were 73 deaths. Kaplan-Meier analysis and Log-rank test showed that overall survival (OS) was significantly shortened in patients with rs7758229 TT as compared with that in those with rs7758229 GG (Log-rank: χ2=11.254, P<0.05). Cox proportional hazard model results showed that rs7758229 mononuclear acid polymorphism [TT type/GG type, P<0.05, odds ratio (OR): 2.357, 95% confidence interval (CI): 1.524-6.317], TNM stage (P<0.05, OR: 1.194, 95%CI: 0.031-3.491), age (P<0.05, OR: 1.354, 95%CI: 1.067-4.357), degree of tumor differentiation (P<0.05, OR: 1.687, 95%CI: 1.108-4.217), and margin situation (P<0.05, OR: 1.947, 95%CI: 1.354-5.218) were independent prognostic factors influencing OS in patients with pancreatic cancer (P<0.05). Conclusion The rs7758229 polymorphism of SLC22A3 gene is associated with the survival rate of pancreatic cancer. The OS of TT patients is shorter. Key words: Solute carrier transporter 22A3 gene; Mononucleotide polymorphism; Pancreatic cancer; Overall survival
{"title":"Correlation between solute carrier transporter 22A3 gene rs7758229 polymorphism and prognosis of pancreatic cancer","authors":"P. Lei, Chaofeng Tang, W. Bu, Songning Yu","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.006","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.006","url":null,"abstract":"Objective \u0000To investigate the relationship between solute carrier transporter 22A3 (SLC22A3) gene rs7758229 polymorphism and prognosis of pancreatic cancer. \u0000 \u0000 \u0000Methods \u0000One hundred patients with resectable pancreatic cancer who underwent surgery were admitted to our hospital from March 30, 2014 to March 30, 2016 were selected. There were were 61 male and 39 female patients, aging from 35 to 79 years with a median age of (49.1±5.7) years. The location included head and neck of the pancreas (n=66), body and tail of pancreas (n=34). According to the 7th American Joint Committee on Cancer (AJCC) Pancreatic Cancer Staging System, 19 cases were in stage I, 56 cases were in stage Ⅱ and 25 cases in stage Ⅲ. On the 2nd day after admission, 5 ml of fasting venous blood was taken, and the polymorphism of SLC22A3 gene rs7758229 was detected by DNA extraction kit and sequencing method. The patients were followed up and the end point was all-cause death. The correlation between polymorphism of rs7758229 and the prognosis of pancreatic cancer, and the factors affecting the overall survival (OS) of the patients were analyzed. The date analysis was conducted by SPSS 25.0; the gene type distribution deviation was tested by Hardy-Weinberg equilibrium, the correlation analysis of clinical date was analyzed by χ2 test analysis. The correlation between polymorphism of rs7758229 and the prognosis of pancreatic cancer, and the factors affecting the OS of the patients were analyzed by by Kaplan- Meier method and Cox model respectively. \u0000 \u0000 \u0000Results \u0000There were 28 cases (28.0%) of GG type, 51 cases (51.0%) of GT type, and 21 cases (21.0%) of TT type. The rs7758229 mononucleic acid polymorphism was significantly correlated with the degree of tumor differentiation and clinical stage (χ2=10.209, 10.826, P<0.05). Six patients were lost to follow-up (2 patients with GG, 3 patients with GT, and 1 patient with TT) with a median follow-up of 46 months. There were 73 deaths. Kaplan-Meier analysis and Log-rank test showed that overall survival (OS) was significantly shortened in patients with rs7758229 TT as compared with that in those with rs7758229 GG (Log-rank: χ2=11.254, P<0.05). Cox proportional hazard model results showed that rs7758229 mononuclear acid polymorphism [TT type/GG type, P<0.05, odds ratio (OR): 2.357, 95% confidence interval (CI): 1.524-6.317], TNM stage (P<0.05, OR: 1.194, 95%CI: 0.031-3.491), age (P<0.05, OR: 1.354, 95%CI: 1.067-4.357), degree of tumor differentiation (P<0.05, OR: 1.687, 95%CI: 1.108-4.217), and margin situation (P<0.05, OR: 1.947, 95%CI: 1.354-5.218) were independent prognostic factors influencing OS in patients with pancreatic cancer (P<0.05). \u0000 \u0000 \u0000Conclusion \u0000The rs7758229 polymorphism of SLC22A3 gene is associated with the survival rate of pancreatic cancer. The OS of TT patients is shorter. \u0000 \u0000 \u0000Key words: \u0000Solute carrier transporter 22A3 gene; Mononucleotide polymorphism; Pancreatic cancer; Overall survival","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"22-24"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43132078","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.019
Shifeng Yang, Lei Wang, Xiao Ma, Mingqi Yang
Objective To investigate the effect of long non-coding RNA maternally expressed gene 3 (MEG3) on the proliferation and invasion capacity of glioma cells through the Wnt/human-catenin signal pathway. Methods Glioma cells were divided into 3 groups: the blank control group (CON), the gene transfection group (MEG3) and the gene inhibition group (siMEG3). The cells in blank group were left untreated and cultured normally. MEG3 gene was transfected in the gene transfection group, and gene interference was conducted in the gene inhibition group. Protein expression was determined by Western blotting, and gene expression was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). Results After transfection of MEG3 gene, the apoptosis rate increased to 61.05±2.77 with the prolongation of culture time, which was higher than that of the control group and the gene inhibition group (F=23.543, P<0.05). After MEG3 gene knockout, the apoptosis rate of glioma cells was higher than that of the control group (P<0.05). After 48 hours of transfection, the number of cell migration was 105.36±15.27 (F=33.350, P<0.05). When MEG3 gene was inhibited, the number of cell migration was 989.41±11.06, higher than that of the control group (F=40.667, P<0.05). After MEG3 gene transfection, the number of invasion cells was 251.25±35.85, which was lower than that of the control group and gene knockout group (F=31.167, P<0.05). After MEG3 gene was knocked out, the number of invasion cells was 1 500.00± 84.76, which was higher than that of the blank control group (F=63.762, P<0.05). Compared with the gene inhibition group, the c-Jun gene of the control group and the gene transfection group decreased, and the gene transfection group was lower than the blank control group (F=15.426, P<0.05). Conclusion Long non-coding RNA MEG3 can inhibit the proliferation and invasion of glioma cells through the Wnt/human-catenin signal pathway. Key words: Glioma; Protein; Gene
{"title":"Effect of long non-coding RNA maternally expressed gene 3 on proliferation and invasion capacity of glioma cells through the Wnt/human-catenin signal pathway","authors":"Shifeng Yang, Lei Wang, Xiao Ma, Mingqi Yang","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.019","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.019","url":null,"abstract":"Objective \u0000To investigate the effect of long non-coding RNA maternally expressed gene 3 (MEG3) on the proliferation and invasion capacity of glioma cells through the Wnt/human-catenin signal pathway. \u0000 \u0000 \u0000Methods \u0000Glioma cells were divided into 3 groups: the blank control group (CON), the gene transfection group (MEG3) and the gene inhibition group (siMEG3). The cells in blank group were left untreated and cultured normally. MEG3 gene was transfected in the gene transfection group, and gene interference was conducted in the gene inhibition group. Protein expression was determined by Western blotting, and gene expression was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). \u0000 \u0000 \u0000Results \u0000After transfection of MEG3 gene, the apoptosis rate increased to 61.05±2.77 with the prolongation of culture time, which was higher than that of the control group and the gene inhibition group (F=23.543, P<0.05). After MEG3 gene knockout, the apoptosis rate of glioma cells was higher than that of the control group (P<0.05). After 48 hours of transfection, the number of cell migration was 105.36±15.27 (F=33.350, P<0.05). When MEG3 gene was inhibited, the number of cell migration was 989.41±11.06, higher than that of the control group (F=40.667, P<0.05). After MEG3 gene transfection, the number of invasion cells was 251.25±35.85, which was lower than that of the control group and gene knockout group (F=31.167, P<0.05). After MEG3 gene was knocked out, the number of invasion cells was 1 500.00± 84.76, which was higher than that of the blank control group (F=63.762, P<0.05). Compared with the gene inhibition group, the c-Jun gene of the control group and the gene transfection group decreased, and the gene transfection group was lower than the blank control group (F=15.426, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000Long non-coding RNA MEG3 can inhibit the proliferation and invasion of glioma cells through the Wnt/human-catenin signal pathway. \u0000 \u0000 \u0000Key words: \u0000Glioma; Protein; Gene","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"67-70"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45718825","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective To investigate the regulatory effect of mechanical vibration intervention on endocrine hormone fibroblast growth factor23 (FGF23) in ovariectomized fracture rats. Methods A model of postmenopausal osteoporosis was established in 45 female rats, and the animals were randomly divided into three groups: Sham operation group (Sham, n=15), ovariectomized fracture model group (OVX, n=15) and ovariectomized fracture vibration group (OVX-V, n=15). The rats in OVX-V group were subjected to whole-body vibration at a frequency of 35 Hz, a vibration amplitude of 2 mm, and an acceleration of 0.5 g, 20 min/d and 5 t/w, 5 days after fracture surgery. The rats in Sham group and OVX group were placed on the vibrating table for 20 min at 5th day after the operation, and no vibration was performed. Western blotting was used to detect the expression of FGF23 protein in the fracture end of rats. SPSS 19.0 statistical software was used to analyze the results. The results were expressed as mean±standard deviation (Mean±SD). Comparisons between groups were analyzed by one-way ANOVA. Results The protein expression of FGF23 in the fracture end of OVX-V group was significantly lower than that in OVX group at 2nd, 4th and 6th week [(0.846±0.012, 1.315±0.021, 1.315±0.021) vs. (0.703±0.009, 0.466±0.011, 0.380±0.005), P<0.01]. . The protein expression of FGF23 in the fracture end of OVX group was significantly higher than that in Sham group at 2nd, 4th and 6th week [(0.168±0.004, 0.321±0.003, 0.417±0.009) vs. (0.846±0.012, 1.315±0.021, 1.315±0.021), P<0.01), showing an increasing trend, suggesting that mechanical vibration can reduce the expression level of FGF23 in the bone tissue of oarotomy rats. Conclusion Mechanical vibration can reduce the expression of FGF23 in bone tissue, promote osteoblast differentiation, regulate bone mineralization and promote postmenopausal osteoporosis fracture healing. Key words: Mechanical vibration; Osteoporosis; Fracture; Endocrine hormones; Fibroblast growth factor23
{"title":"Regulatory effect of mechanical vibration on fibroblast growth factor 23 antibody during fracture healing in ovariectomized rats","authors":"Xue-hong Wang, Daming Sun, Dandan Zhou, Tingting Li, Yi-Ze Wang, Yongliang Hong, Kai Cheng","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.034","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.034","url":null,"abstract":"Objective \u0000To investigate the regulatory effect of mechanical vibration intervention on endocrine hormone fibroblast growth factor23 (FGF23) in ovariectomized fracture rats. \u0000 \u0000 \u0000Methods \u0000A model of postmenopausal osteoporosis was established in 45 female rats, and the animals were randomly divided into three groups: Sham operation group (Sham, n=15), ovariectomized fracture model group (OVX, n=15) and ovariectomized fracture vibration group (OVX-V, n=15). The rats in OVX-V group were subjected to whole-body vibration at a frequency of 35 Hz, a vibration amplitude of 2 mm, and an acceleration of 0.5 g, 20 min/d and 5 t/w, 5 days after fracture surgery. The rats in Sham group and OVX group were placed on the vibrating table for 20 min at 5th day after the operation, and no vibration was performed. Western blotting was used to detect the expression of FGF23 protein in the fracture end of rats. SPSS 19.0 statistical software was used to analyze the results. The results were expressed as mean±standard deviation (Mean±SD). Comparisons between groups were analyzed by one-way ANOVA. \u0000 \u0000 \u0000Results \u0000The protein expression of FGF23 in the fracture end of OVX-V group was significantly lower than that in OVX group at 2nd, 4th and 6th week [(0.846±0.012, 1.315±0.021, 1.315±0.021) vs. (0.703±0.009, 0.466±0.011, 0.380±0.005), P<0.01]. . The protein expression of FGF23 in the fracture end of OVX group was significantly higher than that in Sham group at 2nd, 4th and 6th week [(0.168±0.004, 0.321±0.003, 0.417±0.009) vs. (0.846±0.012, 1.315±0.021, 1.315±0.021), P<0.01), showing an increasing trend, suggesting that mechanical vibration can reduce the expression level of FGF23 in the bone tissue of oarotomy rats. \u0000 \u0000 \u0000Conclusion \u0000Mechanical vibration can reduce the expression of FGF23 in bone tissue, promote osteoblast differentiation, regulate bone mineralization and promote postmenopausal osteoporosis fracture healing. \u0000 \u0000 \u0000Key words: \u0000Mechanical vibration; Osteoporosis; Fracture; Endocrine hormones; Fibroblast growth factor23","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"118-120"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45960440","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.038
C. Hong, Jingjing Fan, B. Ma
Objective To observe the expression of vesicular associated membrane protein 5 (VAMP5) in triple negative breast cancer (TNBC) and its relationship with clinicopathological features and prognosis of TNBC. Methods The expression of VAMP5 was detected by immunohistochemistry, and that in MDA-MB-231 cells and MCF-10A cells was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). The relationship between the expression of VAMP5 and clinicopathological parameters of TNBC patients was analyzed by SPSS 25.0 statistical software, and the survival of TNBC patients was analyzed by Kaplan-Meier method. Results As compared with normal breast cells, the expression level of VAMP5 in TNBC cells was significantly reduced (P 0.05). Conclusion The low expression of VAMP5 in TNBC tissue is negatively correlated with the malignant degree of TNBC. Key words: Triple negative breast cancer; Vesicular associated membrane protein 5; Soluble NSF attachment protein receptors
{"title":"Expression and clinical significance of vesicular associated membrane protein 5 in triple negative breast cancer","authors":"C. Hong, Jingjing Fan, B. Ma","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.038","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.038","url":null,"abstract":"Objective \u0000To observe the expression of vesicular associated membrane protein 5 (VAMP5) in triple negative breast cancer (TNBC) and its relationship with clinicopathological features and prognosis of TNBC. \u0000 \u0000 \u0000Methods \u0000The expression of VAMP5 was detected by immunohistochemistry, and that in MDA-MB-231 cells and MCF-10A cells was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). The relationship between the expression of VAMP5 and clinicopathological parameters of TNBC patients was analyzed by SPSS 25.0 statistical software, and the survival of TNBC patients was analyzed by Kaplan-Meier method. \u0000 \u0000 \u0000Results \u0000As compared with normal breast cells, the expression level of VAMP5 in TNBC cells was significantly reduced (P 0.05). \u0000 \u0000 \u0000Conclusion \u0000The low expression of VAMP5 in TNBC tissue is negatively correlated with the malignant degree of TNBC. \u0000 \u0000 \u0000Key words: \u0000Triple negative breast cancer; Vesicular associated membrane protein 5; Soluble NSF attachment protein receptors","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"131-133"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45749779","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.002
Cheng-xi Liu, Yi-zhi Wang, Junchao Guo
With insidious onset and dismal prognosis, mortality of pancreatic cancer is very high, and incidence in younger patients of China is increasing. Early detection and specific treatment are able to modify the prognosis and prolong the survival of patients. So exploration of new diagnostic method and development of novel therapy target are still the key points in research of pancreatic cancer. As an important part of non-coding RNA, circular RNA (circRNA) has been proven to be involved in the genesis, progress and prognosis. When concerned as diagnostic marker and pharmaceutic target, circRNA has more advantages than others for its stable structure, extensive distribution and specificity of tissue. Because of the connection with exosomes, the researches of exosome make a big effort on translation of circRNA in the clinical management of malignant tumors. This paper will review the latest progress of circRNA and focus on the current research and possible future application of circRNA in pancreatic cancer. Key words: Pancreatic cancer; Circular RNA; Exosome
{"title":"Current progress of circular RNA and the application in pancreatic cancer","authors":"Cheng-xi Liu, Yi-zhi Wang, Junchao Guo","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.002","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.002","url":null,"abstract":"With insidious onset and dismal prognosis, mortality of pancreatic cancer is very high, and incidence in younger patients of China is increasing. Early detection and specific treatment are able to modify the prognosis and prolong the survival of patients. So exploration of new diagnostic method and development of novel therapy target are still the key points in research of pancreatic cancer. As an important part of non-coding RNA, circular RNA (circRNA) has been proven to be involved in the genesis, progress and prognosis. When concerned as diagnostic marker and pharmaceutic target, circRNA has more advantages than others for its stable structure, extensive distribution and specificity of tissue. Because of the connection with exosomes, the researches of exosome make a big effort on translation of circRNA in the clinical management of malignant tumors. This paper will review the latest progress of circRNA and focus on the current research and possible future application of circRNA in pancreatic cancer. \u0000 \u0000Key words: \u0000Pancreatic cancer; Circular RNA; Exosome","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"5-11"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43366321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective To investigate the effect and mechanism of irisin postconditioning on hepatic ischemia reperfusion injury in rats. Methods Totally 36 SD rats were randomly divided into sham group (S), model group (I/R) and irisin group (Ir), with 12 rats in each group. Hepatic ischemia was constructed in model group and irisin group by ligating left middle lobe of liver vascular branches for 60 min. Irisin was administered at the beginning of reperfusion by intravenous injection at the concentration of 10 μg/kg for rats in irisin group. We only dissected the hepatic duodenal ligament of rats in the S group. Serum and liver tissues were collected at 6 h after reperfusion through the inferior chamber. The serum levels of alanine aminotransferase (ALT), glutamic oxalacetic transaminase (AST), interleukin (IL)-6, IL-1βand tumor necrosis factor-α (TNF-α) were examined. The contents of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GPX) in liver tissue were measured. The pathological changes of liver tissue were observed by hematoxylin and eosin (HE) staining. The phosphorylations of janus kinase 2 (JAK2), signal transducer and activators of transcription 3 (STAT3) and B cell lymphoma/leukemia-2 (bcl-2), bcl-2 associated X protein (bax), cysteinyl aspartate-specific protease (Caspase)-3 were assessed by Western blotting. Results The ALT levels in S group, I/R group and irisin group were as follows: (75.24±2.65), (705.33±4.02), (385.46±3.58) U/L; The AST levels were as follows: (122.33±6.76), (1 357.54±5.23), (738.26±3.98) U/L; The IL-6 levels were as follows: (104±16), (586±86), (275±35) ng/L; The TNF-α levels were as follows: (92±10), (1 165±102), (511±73) ng/L; The IL-1β levels were as follows: (98±12), (610±79), (312±41) ng/L; The MDA contents were as follows: (174±38), (1 900±460), (1 055±266) ng/L; The SOD contents were as follows: (124±10), (46±8), (70±10) ng/L; The GPX contents were as follows: (106±12), (42±5), (62±11) ng/L; The Caspase-3 levels were as follows: (0.22±0.06), (0.86±0.14), (0.57±0.11) ng/L; The p-JAK2 levels were as follows: 0.44±0.05, 0.91±0.07, 0.62±0.11; The p-STAT3 levels were as follows: 0.35±0.04, 0.86±0.08, 0.57±0.07. As compared with the sham group, the levels of ALT, AST, IL-6, TNF-α, IL-1β, MDA and Caspase-3 were significantly increased (t=445.551, 219.362, 21.700, 7.700, 36.182, 14.132, 24.191, 10.111, 13.753, 7.023, 14.456 and 6.556, P<0.01). The activities of GPX and SOD in model group and irisin group were significantly decreased (t=20.374, 14.104, 15.945 and 10.965, P<0.01), and the pathological damage worsened and the expression levels of activated p-JAK2 and p-STAT3were up-regulated in other groups (t=14.289, 5.479, 19.054 and 8.224, P<0.01); As compared with model group, the levels of ALT, AST, IL-6, TNF-α, IL-1β, MDA and Caspase-3 in irisin group were significantly decreased (t=226.183, 14.000, 14.081, 22.052 and 7.906, P<0.01), and pathological damage was alleviated and th
{"title":"Effect of irisin postconditioning on hepatic ischemia reperfusion injury in rats","authors":"Xin Yang, Huisheng Wu, Hua-qiao Wang, Tie-cheng Yang, Danwen Wang, Li-jie Yang, Maohui Feng, Yanlin Wang","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.016","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.016","url":null,"abstract":"Objective \u0000To investigate the effect and mechanism of irisin postconditioning on hepatic ischemia reperfusion injury in rats. \u0000 \u0000 \u0000Methods \u0000Totally 36 SD rats were randomly divided into sham group (S), model group (I/R) and irisin group (Ir), with 12 rats in each group. Hepatic ischemia was constructed in model group and irisin group by ligating left middle lobe of liver vascular branches for 60 min. Irisin was administered at the beginning of reperfusion by intravenous injection at the concentration of 10 μg/kg for rats in irisin group. We only dissected the hepatic duodenal ligament of rats in the S group. Serum and liver tissues were collected at 6 h after reperfusion through the inferior chamber. The serum levels of alanine aminotransferase (ALT), glutamic oxalacetic transaminase (AST), interleukin (IL)-6, IL-1βand tumor necrosis factor-α (TNF-α) were examined. The contents of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GPX) in liver tissue were measured. The pathological changes of liver tissue were observed by hematoxylin and eosin (HE) staining. The phosphorylations of janus kinase 2 (JAK2), signal transducer and activators of transcription 3 (STAT3) and B cell lymphoma/leukemia-2 (bcl-2), bcl-2 associated X protein (bax), cysteinyl aspartate-specific protease (Caspase)-3 were assessed by Western blotting. \u0000 \u0000 \u0000Results \u0000The ALT levels in S group, I/R group and irisin group were as follows: (75.24±2.65), (705.33±4.02), (385.46±3.58) U/L; The AST levels were as follows: (122.33±6.76), (1 357.54±5.23), (738.26±3.98) U/L; The IL-6 levels were as follows: (104±16), (586±86), (275±35) ng/L; The TNF-α levels were as follows: (92±10), (1 165±102), (511±73) ng/L; The IL-1β levels were as follows: (98±12), (610±79), (312±41) ng/L; The MDA contents were as follows: (174±38), (1 900±460), (1 055±266) ng/L; The SOD contents were as follows: (124±10), (46±8), (70±10) ng/L; The GPX contents were as follows: (106±12), (42±5), (62±11) ng/L; The Caspase-3 levels were as follows: (0.22±0.06), (0.86±0.14), (0.57±0.11) ng/L; The p-JAK2 levels were as follows: 0.44±0.05, 0.91±0.07, 0.62±0.11; The p-STAT3 levels were as follows: 0.35±0.04, 0.86±0.08, 0.57±0.07. As compared with the sham group, the levels of ALT, AST, IL-6, TNF-α, IL-1β, MDA and Caspase-3 were significantly increased (t=445.551, 219.362, 21.700, 7.700, 36.182, 14.132, 24.191, 10.111, 13.753, 7.023, 14.456 and 6.556, P<0.01). The activities of GPX and SOD in model group and irisin group were significantly decreased (t=20.374, 14.104, 15.945 and 10.965, P<0.01), and the pathological damage worsened and the expression levels of activated p-JAK2 and p-STAT3were up-regulated in other groups (t=14.289, 5.479, 19.054 and 8.224, P<0.01); As compared with model group, the levels of ALT, AST, IL-6, TNF-α, IL-1β, MDA and Caspase-3 in irisin group were significantly decreased (t=226.183, 14.000, 14.081, 22.052 and 7.906, P<0.01), and pathological damage was alleviated and th","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"56-59"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48998837","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective To observe the effects of exosomal circular RNA (circRNA)-100338 derived from hepatocellular carcinoma (HCC) on angiogenesis of human umbilical vein endothelial cells (HUVEC). Methods The exosomes from the supernatant of MHCC97H cells were extracted, purified and identified, and co-incubated with HUVECs (37 ℃, 5% CO2). The exosome tracer experiments traced whether the exosomes could be transformed into HUVECs. MHCC97H cells were transfected with Lipofectamine™ 2000 transfection reagent to extract exosomes from the supernatant of HCC cells after interfering with circRNA-100338 (1 g/L). Proliferation assay was used to detect the changes of proliferation ability of HUVECs [The absorbance (A) values were measured at a wavelength of 450 nm], and angiogenesis assay was applied to evaluate the tubular formation of HUVECs (×100). Results The results of transmission electron microscopy showed that most exosomes from MHCC97H cells were round or oval, and the peak size distribution was 120 nm, which accorded with the characteristics of exosome size. Western blotting was used to detect exosome marker proteins. MHCC97H cells-derived exosomes were internalized by HUVECs. The A values of HUVECs co-cultured with circRNA-100338 interfering group and control group were 0.358±0.005 and 0.436±0.011 (t=-16.227, P<0.01), 0.661±0.052 and 0.888±0.031 (t =-9.146, P<0.01), 1.191±0.084 and 1.542±0.038 (t=-9.296, P<0.01) at 48, 72 and 96 h, respectively. The proliferation ability of HUVECs in interfering group was significantly lower than that in control group after 48-h treatment. In the interferening group, the ability of HUVECs to form tubules was decreased, and the tubular nodules were thin and small. Conclusion HCC-derived exosomal circRNA-100338 enhanced the proliferation ability of HUVECs and promoted endothelial angiogenesis. Key words: Carcinoma, hepatocellular; Exosome; Circular RNA-100338; Proliferation; Angiogenesis
{"title":"Effects of exosomal circular RNA-100338 derived from hepatocellular carcinoma on angiogenesis of human umbilical vein endothelial cells","authors":"Xiu-Yan Huang, Zi-Li Huang, Yong Hua Xu, Xin-Yu Huang, Jian Zhou, Zhao-You Tang","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.012","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.012","url":null,"abstract":"Objective \u0000To observe the effects of exosomal circular RNA (circRNA)-100338 derived from hepatocellular carcinoma (HCC) on angiogenesis of human umbilical vein endothelial cells (HUVEC). \u0000 \u0000 \u0000Methods \u0000The exosomes from the supernatant of MHCC97H cells were extracted, purified and identified, and co-incubated with HUVECs (37 ℃, 5% CO2). The exosome tracer experiments traced whether the exosomes could be transformed into HUVECs. MHCC97H cells were transfected with Lipofectamine™ 2000 transfection reagent to extract exosomes from the supernatant of HCC cells after interfering with circRNA-100338 (1 g/L). Proliferation assay was used to detect the changes of proliferation ability of HUVECs [The absorbance (A) values were measured at a wavelength of 450 nm], and angiogenesis assay was applied to evaluate the tubular formation of HUVECs (×100). \u0000 \u0000 \u0000Results \u0000The results of transmission electron microscopy showed that most exosomes from MHCC97H cells were round or oval, and the peak size distribution was 120 nm, which accorded with the characteristics of exosome size. Western blotting was used to detect exosome marker proteins. MHCC97H cells-derived exosomes were internalized by HUVECs. The A values of HUVECs co-cultured with circRNA-100338 interfering group and control group were 0.358±0.005 and 0.436±0.011 (t=-16.227, P<0.01), 0.661±0.052 and 0.888±0.031 (t =-9.146, P<0.01), 1.191±0.084 and 1.542±0.038 (t=-9.296, P<0.01) at 48, 72 and 96 h, respectively. The proliferation ability of HUVECs in interfering group was significantly lower than that in control group after 48-h treatment. In the interferening group, the ability of HUVECs to form tubules was decreased, and the tubular nodules were thin and small. \u0000 \u0000 \u0000Conclusion \u0000HCC-derived exosomal circRNA-100338 enhanced the proliferation ability of HUVECs and promoted endothelial angiogenesis. \u0000 \u0000 \u0000Key words: \u0000Carcinoma, hepatocellular; Exosome; Circular RNA-100338; Proliferation; Angiogenesis","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"40-43"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48132791","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.028
Bo Xue, Jianpeng Hu, Xiaofei Zhang, Y. Wan, Xuechao Xu, F. Cui
Objective To investigate the relationship between the fluctuation of dihydrotestosterone concentration and the expression of mutL homolog 1 (MLH1) in mismatch repair system in prostate epithelial cells (RWPE-2 cell line) and its mechanism. Methods RWPE-2 cells were randomly divided into blank group [anhydrous ethanol solvent and dihydrotestosterone (DHT) concentration of 0 nmol/L], constant concentration group (concentration of DHT: 1, 10 nmol/L) and fluctuating concentration group (concentration of DHT: 1, 10 nmol/L, fluctuating every 24 h). After different time (0, 24, 48 and 72 h), cell counting kit-8 was use to examine cell proliferation and the target genes in androgen receptor (AR) signaling pathway [prostate specific antigen (PSA), FK506 binding protein 51 (FKBP5)], protein kinase B (Akt) signaling pathway [Cyclin D1, B cell lymphoma/leukemia-2 (bcl-2)] and MLH1 was detected by Western blotting, real-time quantitative polymerase chain reaction (Real-time PCR) and immunofluorescence. Graph Pad 7.0 statistical software was used for analysis and differences between groups were analyzed by t test. Results The morphological changes of RWPE-2 cells had a certain concentration-dependent relationship with DHT: constant group and fluctuating group had more cells and better growth than blank group. After RWPE-2 cells were stimulated with DHT for 72 h, as compared with the blank group (6.904±0.143), the relative A values of the constant group [(7.073±0.103) and (8.508±0.187)] and the fluctuation group [(8.662±0.327) and (9.239±0.167)] increased statistically (t=6.805, 4.922, 10.6, P<0.01). Western blotting and RT-qPCR results confirmed that the mRNA and protein levels of every target gene were increased significantly in constant group and fluctuating group (P<0.05) as compared with blank group. Meanwhile, the difference in fluctuating group was more significant than constant group. Whereas, FKBP5 mRNA decreased in fluctuating group as compared with constant group (t=7.101, 10.760, 8.289, 8.088, 8.519, 9.157, 11.330, P<0.05). The distribution of above genes increased in the nucleus after DHT intervention with more significant difference in fluctuating group. Conclusion DHT can up-regulate the expression of the mismatch repair protein MLH1 in RWPE-2 cells, meanwhile, DHT fluctuations can increase this trend and AR and Akt signaling pathways-mediated proliferation is one of the mechanisms. Key words: Dihydrotestosterone fluctuation; Cell proliferation; Mismatch repair; Androgen receptor signal pathway; Protein kinase B signal pathway
目的探讨前列腺上皮细胞错配修复系统中二氢睾酮浓度波动与mutL同源物1（MLH1）表达的关系及其机制。方法将RWPE-2细胞随机分为空白组[无水乙醇溶剂和二氢睾酮（DHT）浓度为0nmol/L]、恒定浓度组（DHT浓度为1,10nmol/L）和波动浓度组（浓度为1,110nmol/L，每24小时波动一次）。在不同时间（0、24、48和72小时）后，使用细胞计数试剂盒-8检测细胞增殖，并通过Western印迹检测雄激素受体（AR）信号通路[前列腺特异性抗原（PSA）、FK506结合蛋白51（FKBP5）]、蛋白激酶B（Akt）信号通路[Cyclin D1、B细胞淋巴瘤/白血病-2（bcl-2）]和MLH1中的靶基因，实时定量聚合酶链反应（real-time PCR）和免疫荧光。使用Graph Pad 7.0统计软件进行分析，并通过t检验分析各组之间的差异。结果RWPE-2细胞的形态学变化与DHT有一定的浓度依赖关系：恒定组和波动组的细胞数和生长情况均高于空白组。DHT刺激RWPE-2细胞72 h后与空白组（6.904±0.143）相比，恒定组[（7.073±0.103）和（8.508±0.187）]和波动组[（8.662±0.327）和（9.239±0.167）]的相对A值均有统计学意义（t=6.805，4.922，10.6，P<0.01）空白组。同时，波动组的差异比恒定组更显著。而FKBP5mRNA在波动组较恒定组降低（t=7.101，10.760，8.289，8.088，8.519，9.157，11.330，P<0.05），DHT干预后上述基因在细胞核中的分布增加，波动组差异更显著。结论DHT可上调RWPE-2细胞错配修复蛋白MLH1的表达，同时DHT的波动可增加这种趋势，AR和Akt信号通路介导的增殖是其机制之一。关键词：双氢睾酮波动；细胞增殖；错配修复；雄激素受体信号通路；蛋白激酶B信号通路
{"title":"Effect of dihydrotestosterone fluctuation on mismatch repair gene mutL homolog 1 in prostate epithelial cells and its mechanism","authors":"Bo Xue, Jianpeng Hu, Xiaofei Zhang, Y. Wan, Xuechao Xu, F. Cui","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.028","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.028","url":null,"abstract":"Objective \u0000To investigate the relationship between the fluctuation of dihydrotestosterone concentration and the expression of mutL homolog 1 (MLH1) in mismatch repair system in prostate epithelial cells (RWPE-2 cell line) and its mechanism. \u0000 \u0000 \u0000Methods \u0000RWPE-2 cells were randomly divided into blank group [anhydrous ethanol solvent and dihydrotestosterone (DHT) concentration of 0 nmol/L], constant concentration group (concentration of DHT: 1, 10 nmol/L) and fluctuating concentration group (concentration of DHT: 1, 10 nmol/L, fluctuating every 24 h). After different time (0, 24, 48 and 72 h), cell counting kit-8 was use to examine cell proliferation and the target genes in androgen receptor (AR) signaling pathway [prostate specific antigen (PSA), FK506 binding protein 51 (FKBP5)], protein kinase B (Akt) signaling pathway [Cyclin D1, B cell lymphoma/leukemia-2 (bcl-2)] and MLH1 was detected by Western blotting, real-time quantitative polymerase chain reaction (Real-time PCR) and immunofluorescence. Graph Pad 7.0 statistical software was used for analysis and differences between groups were analyzed by t test. \u0000 \u0000 \u0000Results \u0000The morphological changes of RWPE-2 cells had a certain concentration-dependent relationship with DHT: constant group and fluctuating group had more cells and better growth than blank group. After RWPE-2 cells were stimulated with DHT for 72 h, as compared with the blank group (6.904±0.143), the relative A values of the constant group [(7.073±0.103) and (8.508±0.187)] and the fluctuation group [(8.662±0.327) and (9.239±0.167)] increased statistically (t=6.805, 4.922, 10.6, P<0.01). Western blotting and RT-qPCR results confirmed that the mRNA and protein levels of every target gene were increased significantly in constant group and fluctuating group (P<0.05) as compared with blank group. Meanwhile, the difference in fluctuating group was more significant than constant group. Whereas, FKBP5 mRNA decreased in fluctuating group as compared with constant group (t=7.101, 10.760, 8.289, 8.088, 8.519, 9.157, 11.330, P<0.05). The distribution of above genes increased in the nucleus after DHT intervention with more significant difference in fluctuating group. \u0000 \u0000 \u0000Conclusion \u0000DHT can up-regulate the expression of the mismatch repair protein MLH1 in RWPE-2 cells, meanwhile, DHT fluctuations can increase this trend and AR and Akt signaling pathways-mediated proliferation is one of the mechanisms. \u0000 \u0000 \u0000Key words: \u0000Dihydrotestosterone fluctuation; Cell proliferation; Mismatch repair; Androgen receptor signal pathway; Protein kinase B signal pathway","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"97-100"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45401240","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective To investigate the effects of rosuvastatin on proliferation, osteogenic differentiation and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) expression of mesenchymal stem cells (MSCs). Methods MSCs were isolated. Rosuvastatin at concentrations of 1×10-11, 1×10-9 and 1×10-7 mol/L were added to the experimental groups, and dimethylsurfoxide (DMSO) served as control. Cell proliferation was measured by methyl thiazol tetrazolium (MTT) assay. After 3, 7, 14 and 21 days of osteogenic induction, alkaline phosphatase (ALP) quantitative analysis, alizarin red staining and quantitative analysis, real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting were performed. T-test was used for comparison between two groups, and ANOVA for more than two groups. Results At 24 h and 48 h, the proliferation of MSCs in high concentration group was significantly higher than in the control group (24 h: 3.57±0.24 vs. 3.14±0.14, t=-3.851, P<0.05; 48 h: 4.19±0.18 vs. 3.44±0.11, t=-6.780, P<0.05). Until 72 h, the proliferation with 1×10-9 mol/L rosuvastatin was also significantly increased (5.42±0.13 vs. 4.47±0.16, t=-8.700, P<0.05). At 3rd day after induction culture, 1×10-7 mol/L rosuvastatin significantly enhanced the genes expression of BMP2, Runx 2, OPN 2, OPN and ColⅠ (BMP2: 2.1±0.2 vs. 1.0±0.2, t=-6.736, P<0.05; Runx2: 5.2±0.6 vs. 1.0±0.1, t=-11.959, P<0.05; OPN: 1.8±0.4 vs. 1.0±0.2, t=-3.098, P<0.05; ColⅠ: 1.9±0.3 vs. 1.0±0.3, t=-3.674, P<0.05); 1×10-7 mol/L rosuvastatin could increase the expression of p-Akt/PI3K. The ALP activity after adding 1×10-9 or 1×10-7 mol/L rosuvastatin was significantly increased at 14th and 21st day (14 d: 12.07±1.67, 18.32±2.26 vs. 3.43±0.36, F=7.200, P<0.05; 21 d: 10.74±1.27, 14.71±3.18 vs. 5.76±0.63, F=6.489, P<0.05). The quantitative results of alizarin red were similar to those of the staining. Meanwhile, the quantitative values of alizarin red in the 1×10-9 and 1×10-7 mol/L groups were also significantly increased (14 d: 5.6±0.8, 6.2±1.2 vs. 1.0±0.2, F=5.600, P<0.05; 21 d: 5.8±1.1, 7.1±1.8 vs. 2.4±0.3, F=5.956, P<0.05). Conclusion Rosuvastatin promotes proliferation and osteogenic differentiation of MSCs, which may be related to PI3K/Akt signaling. Key words: Rosuvastatin; Phosphatidylinositol-3-kinase/protein kinase B signaling pathway; Mesenchymal stem cells; Osteogenic differentiation
目的探讨瑞舒伐他汀对间充质干细胞(MSCs)增殖、成骨分化及磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B (Akt)表达的影响。方法分离MSCs。实验组分别加入浓度为1×10-11、1×10-9和1×10-7 mol/L的瑞舒伐他汀，对照组分别加入二甲苏福啶(DMSO)。采用甲基噻唑四氮唑(MTT)法检测细胞增殖。成骨诱导3、7、14、21 d后进行碱性磷酸酶(ALP)定量分析、茜素红染色及定量分析、实时定量聚合酶链反应(real-time PCR)和Western blotting。两组间比较采用t检验，两组以上采用方差分析。结果24 h、48 h，高浓度组MSCs的增殖明显高于对照组(24 h: 3.57±0.24 vs. 3.14±0.14,t=-3.851, P<0.05;48 h: 4.19±0.18和3.44±0.11,t = -6.780, P < 0.05)。1×10-9 mol/L瑞舒伐他汀治疗至72 h时，细胞增殖也显著增加(5.42±0.13∶4.47±0.16,t=-8.700, P<0.05)。诱导培养后第3天，1×10-7 mol/L瑞舒伐他汀显著提高BMP2、runx2、opn2、OPN和Col基因表达Ⅰ(BMP2: 2.1±0.2 vs. 1.0±0.2,t=-6.736, P<0.05;Runx2: 5.2±0.6 vs. 1.0±0.1,t=-11.959, P<0.05;OPN: 1.8±0.4 vs. 1.0±0.2,t=-3.098, P<0.05;ColⅠ:1.9±0.3 vs. 1.0±0.3,t=-3.674, P<0.05);1×10-7 mol/L瑞舒伐他汀可增加p-Akt/PI3K的表达。添加1×10-9或1×10-7 mol/L瑞舒伐他汀后第14天和第21天ALP活性显著升高(14 d: 12.07±1.67,18.32±2.26 vs. 3.43±0.36,F=7.200, P<0.05;21 d: 10.74±1.27,14.71±3.18和5.76±0.63,F = 6.489, P < 0.05)。茜素红的定量结果与染色结果相似。同时，1×10-9和1×10-7 mol/L组的芹素红定量值也显著升高(14 d: 5.6±0.8、6.2±1.2 vs. 1.0±0.2,F=5.600, P<0.05;21 d: 5.8±1.1,7.1±1.8和2.4±0.3,F = 5.956, P < 0.05)。结论瑞舒伐他汀促进MSCs增殖和成骨分化，可能与PI3K/Akt信号通路有关。关键词:瑞舒伐他汀;磷脂酰肌醇-3激酶/蛋白激酶B信号通路;间充质干细胞;成骨分化
{"title":"Rosuvastatin regulates osteogenic differentiation of mesenchymal stem cells through phosphatidylinositol-3-kinase/protein kinase B","authors":"Xuepeng Wang, Chunchun Zou, Changju Hou, Maoqiang Li, Z. Bian, Liulong Zhu","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.029","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.029","url":null,"abstract":"Objective \u0000To investigate the effects of rosuvastatin on proliferation, osteogenic differentiation and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) expression of mesenchymal stem cells (MSCs). \u0000 \u0000 \u0000Methods \u0000MSCs were isolated. Rosuvastatin at concentrations of 1×10-11, 1×10-9 and 1×10-7 mol/L were added to the experimental groups, and dimethylsurfoxide (DMSO) served as control. Cell proliferation was measured by methyl thiazol tetrazolium (MTT) assay. After 3, 7, 14 and 21 days of osteogenic induction, alkaline phosphatase (ALP) quantitative analysis, alizarin red staining and quantitative analysis, real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting were performed. T-test was used for comparison between two groups, and ANOVA for more than two groups. \u0000 \u0000 \u0000Results \u0000At 24 h and 48 h, the proliferation of MSCs in high concentration group was significantly higher than in the control group (24 h: 3.57±0.24 vs. 3.14±0.14, t=-3.851, P<0.05; 48 h: 4.19±0.18 vs. 3.44±0.11, t=-6.780, P<0.05). Until 72 h, the proliferation with 1×10-9 mol/L rosuvastatin was also significantly increased (5.42±0.13 vs. 4.47±0.16, t=-8.700, P<0.05). At 3rd day after induction culture, 1×10-7 mol/L rosuvastatin significantly enhanced the genes expression of BMP2, Runx 2, OPN 2, OPN and ColⅠ (BMP2: 2.1±0.2 vs. 1.0±0.2, t=-6.736, P<0.05; Runx2: 5.2±0.6 vs. 1.0±0.1, t=-11.959, P<0.05; OPN: 1.8±0.4 vs. 1.0±0.2, t=-3.098, P<0.05; ColⅠ: 1.9±0.3 vs. 1.0±0.3, t=-3.674, P<0.05); 1×10-7 mol/L rosuvastatin could increase the expression of p-Akt/PI3K. The ALP activity after adding 1×10-9 or 1×10-7 mol/L rosuvastatin was significantly increased at 14th and 21st day (14 d: 12.07±1.67, 18.32±2.26 vs. 3.43±0.36, F=7.200, P<0.05; 21 d: 10.74±1.27, 14.71±3.18 vs. 5.76±0.63, F=6.489, P<0.05). The quantitative results of alizarin red were similar to those of the staining. Meanwhile, the quantitative values of alizarin red in the 1×10-9 and 1×10-7 mol/L groups were also significantly increased (14 d: 5.6±0.8, 6.2±1.2 vs. 1.0±0.2, F=5.600, P<0.05; 21 d: 5.8±1.1, 7.1±1.8 vs. 2.4±0.3, F=5.956, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000Rosuvastatin promotes proliferation and osteogenic differentiation of MSCs, which may be related to PI3K/Akt signaling. \u0000 \u0000 \u0000Key words: \u0000Rosuvastatin; Phosphatidylinositol-3-kinase/protein kinase B signaling pathway; Mesenchymal stem cells; Osteogenic differentiation","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"101-104"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46317928","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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