Shiyong Ling, Chen Yan, Kai Huang, Bo Lv, Hua Wang, Xiaoyan Wang, Jun Chen, Jingchuan Sun
{"title":"PTEN介导的TGF-β/Smad2通路对骨质疏松性胫骨骨折大鼠成骨分化和骨髓间充质干细胞张力作用的分析","authors":"Shiyong Ling, Chen Yan, Kai Huang, Bo Lv, Hua Wang, Xiaoyan Wang, Jun Chen, Jingchuan Sun","doi":"10.1155/2022/1004203","DOIUrl":null,"url":null,"abstract":"Purpose. To discuss effects of phosphatase and tensin homolog protein (PTEN)-mediated transforming growth factor-β (TGF-β)/Smad homologue 2 (Smad2) pathway on osteogenic differentiation in osteoporotic (OP) tibial fracture rats and bone marrow mesenchymal stem cell (BMSC) under tension. Methods. A tibial fracture model was established. The rats were divided into sham-operated group and model group, and tibia tissue was collected. Purchase well-grown cultured rat BMSC, and use the Flexercell in vitro cell mechanics loading device to apply tension. The expression of PTEN was detected by qRT-PCR. After the BMSCs were transfected with si-PTEN and oe-PTEN, the force was applied to detect cell differentiation. The expression of TGF-β/Smad2 protein was detected by Western blot. The formation of calcium nodules in BMSC was detected by alkaline phosphatase (ALP) staining and alizarin red (AR) staining. Results. The expression of PTEN was higher in the model group and tension MSC group, and the expression of TGF-β and Smad2 protein was lower. The expression of TGF-β and Smad2 protein in oe-PTEN group was lower than the oe-NC group and control group. The expression of TGF-β and Smad2 protein in si-PTEN group was higher than the si-NC group and control group. The results of ALP staining and AR staining also confirmed the above results. Conclusion. PTEN-mediated TGF-β/Smad2 pathway may play a key role in the osteogenic differentiation of OP tibial fracture rats. Downregulation of PTEN and upregulation of TGF-β/Smad2 signal can promote the osteogenic differentiation of BMSC under tension.","PeriodicalId":9844,"journal":{"name":"Cellular Microbiology","volume":" ","pages":""},"PeriodicalIF":2.6000,"publicationDate":"2022-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Analysis of Effects of PTEN-Mediated TGF-β/Smad2 Pathway on Osteogenic Differentiation in Osteoporotic Tibial Fracture Rats and Bone Marrow Mesenchymal Stem Cell under Tension\",\"authors\":\"Shiyong Ling, Chen Yan, Kai Huang, Bo Lv, Hua Wang, Xiaoyan Wang, Jun Chen, Jingchuan Sun\",\"doi\":\"10.1155/2022/1004203\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Purpose. To discuss effects of phosphatase and tensin homolog protein (PTEN)-mediated transforming growth factor-β (TGF-β)/Smad homologue 2 (Smad2) pathway on osteogenic differentiation in osteoporotic (OP) tibial fracture rats and bone marrow mesenchymal stem cell (BMSC) under tension. Methods. A tibial fracture model was established. The rats were divided into sham-operated group and model group, and tibia tissue was collected. Purchase well-grown cultured rat BMSC, and use the Flexercell in vitro cell mechanics loading device to apply tension. The expression of PTEN was detected by qRT-PCR. After the BMSCs were transfected with si-PTEN and oe-PTEN, the force was applied to detect cell differentiation. The expression of TGF-β/Smad2 protein was detected by Western blot. The formation of calcium nodules in BMSC was detected by alkaline phosphatase (ALP) staining and alizarin red (AR) staining. Results. The expression of PTEN was higher in the model group and tension MSC group, and the expression of TGF-β and Smad2 protein was lower. The expression of TGF-β and Smad2 protein in oe-PTEN group was lower than the oe-NC group and control group. The expression of TGF-β and Smad2 protein in si-PTEN group was higher than the si-NC group and control group. The results of ALP staining and AR staining also confirmed the above results. Conclusion. PTEN-mediated TGF-β/Smad2 pathway may play a key role in the osteogenic differentiation of OP tibial fracture rats. Downregulation of PTEN and upregulation of TGF-β/Smad2 signal can promote the osteogenic differentiation of BMSC under tension.\",\"PeriodicalId\":9844,\"journal\":{\"name\":\"Cellular Microbiology\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":2.6000,\"publicationDate\":\"2022-04-29\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Cellular Microbiology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1155/2022/1004203\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"CELL BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cellular Microbiology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1155/2022/1004203","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"CELL BIOLOGY","Score":null,"Total":0}
Analysis of Effects of PTEN-Mediated TGF-β/Smad2 Pathway on Osteogenic Differentiation in Osteoporotic Tibial Fracture Rats and Bone Marrow Mesenchymal Stem Cell under Tension
Purpose. To discuss effects of phosphatase and tensin homolog protein (PTEN)-mediated transforming growth factor-β (TGF-β)/Smad homologue 2 (Smad2) pathway on osteogenic differentiation in osteoporotic (OP) tibial fracture rats and bone marrow mesenchymal stem cell (BMSC) under tension. Methods. A tibial fracture model was established. The rats were divided into sham-operated group and model group, and tibia tissue was collected. Purchase well-grown cultured rat BMSC, and use the Flexercell in vitro cell mechanics loading device to apply tension. The expression of PTEN was detected by qRT-PCR. After the BMSCs were transfected with si-PTEN and oe-PTEN, the force was applied to detect cell differentiation. The expression of TGF-β/Smad2 protein was detected by Western blot. The formation of calcium nodules in BMSC was detected by alkaline phosphatase (ALP) staining and alizarin red (AR) staining. Results. The expression of PTEN was higher in the model group and tension MSC group, and the expression of TGF-β and Smad2 protein was lower. The expression of TGF-β and Smad2 protein in oe-PTEN group was lower than the oe-NC group and control group. The expression of TGF-β and Smad2 protein in si-PTEN group was higher than the si-NC group and control group. The results of ALP staining and AR staining also confirmed the above results. Conclusion. PTEN-mediated TGF-β/Smad2 pathway may play a key role in the osteogenic differentiation of OP tibial fracture rats. Downregulation of PTEN and upregulation of TGF-β/Smad2 signal can promote the osteogenic differentiation of BMSC under tension.
期刊介绍:
Cellular Microbiology aims to publish outstanding contributions to the understanding of interactions between microbes, prokaryotes and eukaryotes, and their host in the context of pathogenic or mutualistic relationships, including co-infections and microbiota. We welcome studies on single cells, animals and plants, and encourage the use of model hosts and organoid cultures. Submission on cell and molecular biological aspects of microbes, such as their intracellular organization or the establishment and maintenance of their architecture in relation to virulence and pathogenicity are also encouraged. Contributions must provide mechanistic insights supported by quantitative data obtained through imaging, cellular, biochemical, structural or genetic approaches.