Pub Date : 2024-08-12DOI: 10.1134/s1990519x24700457
M. V. Khrapova, O. S. Bryushinina, Yu. G. Zyuzkova, N. V. Kandalintseva, E. B. Menshchikova
Abstract
TS-13 (sodium 3-(3'-tert-butyl-4'-hydroxyphenyl)propyl thiosulfonate) is a synthetic antioxidant that in numerous studies has demonstrated biological effectiveness in modeling pathological conditions in vivo, in particular, in the model of Parkinson’s disease. To establish whether these effects are indirect or associated with, among other things, the direct effect of TS-13 on the organs and tissues of animals, in this work we determined the concentration of TS-13 in the blood plasma and brain of rats after intragastric administration. After a single intragastric injection of TS-13 solution at a dose of 100 mg/kg to male Sprague Dawley rats (n = 57), biomaterial (blood, brain) was collected over 24 h. To measure the concentration of a substance in samples, a bioanalytical technique using high-performance liquid chromatography with UV detection was developed and validated. The quantitative determination method was developed by us for the first time and validated before the study. It has been established that the calculated values of the calibration samples meet the acceptance criteria (have the required accuracy and precision) in the concentration range from 0.05 to 6 μg/mL, R = 0.9998. The results of determining the concentration of TS-13 in the blood plasma and brain of rats showed that after a single oral administration the compound enters the blood, where it is detected within 15 h (average retention time 7.94 h, half-life 7.59 h, elimination constant 0.13 h–1, total clearance 40.1 L/(kg h)), and also penetrates the blood–brain barrier, quickly entering the brain (maximum concentration is achieved after 1 h). The compound has low affinity for brain tissue (tissue availability 0.32), and, therefore, its concentration does not reach high values; however, slow elimination of the substance is observed—average retention time 6.56 h, half-life 6.43 h, elimination constant 0.11 h–1. After a single intragastric administration to rats, TS-13 enters the blood, where at least part of it is detected unchanged after 30 min, reaching maximum values after 1 h. A similar kinetics of the substance is characteristic of the brain, where it is found in smaller quantities. Thus, as a result of the study, it was shown that TS-13 penetrates the blood–brain barrier and is able to directly affect brain structures, which, however, does not negate the possibility of an indirect effect, mediated by the ability to change the activity of intra- and intercellular signaling systems.
{"title":"The New Synthetic Monophenolic Antioxidant TS-13 Penetrates the Blood–Brain Barrier","authors":"M. V. Khrapova, O. S. Bryushinina, Yu. G. Zyuzkova, N. V. Kandalintseva, E. B. Menshchikova","doi":"10.1134/s1990519x24700457","DOIUrl":"https://doi.org/10.1134/s1990519x24700457","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>TS-13 (sodium 3-(3'-<i>tert</i>-butyl-4'-hydroxyphenyl)propyl thiosulfonate) is a synthetic antioxidant that in numerous studies has demonstrated biological effectiveness in modeling pathological conditions in vivo, in particular, in the model of Parkinson’s disease. To establish whether these effects are indirect or associated with, among other things, the direct effect of TS-13 on the organs and tissues of animals, in this work we determined the concentration of TS-13 in the blood plasma and brain of rats after intragastric administration. After a single intragastric injection of TS-13 solution at a dose of 100 mg/kg to male Sprague Dawley rats (<i>n</i> = 57), biomaterial (blood, brain) was collected over 24 h. To measure the concentration of a substance in samples, a bioanalytical technique using high-performance liquid chromatography with UV detection was developed and validated. The quantitative determination method was developed by us for the first time and validated before the study. It has been established that the calculated values of the calibration samples meet the acceptance criteria (have the required accuracy and precision) in the concentration range from 0.05 to 6 μg/mL, <i>R</i> = 0.9998. The results of determining the concentration of TS-13 in the blood plasma and brain of rats showed that after a single oral administration the compound enters the blood, where it is detected within 15 h (average retention time 7.94 h, half-life 7.59 h, elimination constant 0.13 h<sup>–1</sup>, total clearance 40.1 L/(kg h)), and also penetrates the blood–brain barrier, quickly entering the brain (maximum concentration is achieved after 1 h). The compound has low affinity for brain tissue (tissue availability 0.32), and, therefore, its concentration does not reach high values; however, slow elimination of the substance is observed—average retention time 6.56 h, half-life 6.43 h, elimination constant 0.11 h<sup>–1</sup>. After a single intragastric administration to rats, TS-13 enters the blood, where at least part of it is detected unchanged after 30 min, reaching maximum values after 1 h. A similar kinetics of the substance is characteristic of the brain, where it is found in smaller quantities. Thus, as a result of the study, it was shown that TS-13 penetrates the blood–brain barrier and is able to directly affect brain structures, which, however, does not negate the possibility of an indirect effect, mediated by the ability to change the activity of intra- and intercellular signaling systems.</p>","PeriodicalId":9705,"journal":{"name":"Cell and Tissue Biology","volume":"22 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141931651","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 : 2024-08-12DOI: 10.1134/s1990519x24700421
S. S. Sharaf, A. Lekshmi, K. Sujathan
Abstract
This study demonstrated that silencing of HIF1A using esiRNA (endoribonuclease prepared small interfering RNA) could inhibit breast cancer cells proliferation, increase the apoptosis rate, and inhibit cell invasion and migration. These results suggested that HIF1A gene may involve in the occurrence and development of triple negative breast cancer phenotype. Hypoxia is an important hallmark of most solid tumors. In breast cancer, hypoxia is more evident in TNBC (triple negative breast cancer) than in other breast cancer subtypes. Intratumoral analysis in TNBC showed that tumor areas with high levels of hypoxia, indicated higher expression of HIF1A. Here we investigated the effect of silencing HIF1A on the biological function of triple negative breast cancer. Reverse transcription-polymerase chain reaction and western blot were used to detect HIF1A expression in different breast cancer cells and patient tissues. esiRNA was used to silence HIF1A in MDA-MB-231 cells. Then, BrdU assay was performed to study cell proliferation. Additionally, cell apoptosis after HIF1A knockdown was measured by Annexin V/propidium iodide staining followed by Caspase assay, and cell migration and invasion was detected by trans-well assay. IHC analysis suggested a strong overexpression of HIF1A in TNBC tissues. Silencing suggested that HIF1A significantly promotes cell proliferation and migration, resulting in metastasis and downregulates apoptosis. MDA-MB-231 cells in which HIF1A expression was inhibited were less invasive, with reduced resistance to hypoxia, impaired migration, increased apoptosis and reduced capacity to cause metastasis. HIF1A may be a dominant factor driving the metastatic progression and apoptosis of triple negative breast cancer and can be a potent therapeutic target for the disease.
{"title":"esiRNA Mediated Silencing of HIF1A Regulates Migration, Invasion, Apoptosis, and Proliferation of MDA-MB-231 Cells","authors":"S. S. Sharaf, A. Lekshmi, K. Sujathan","doi":"10.1134/s1990519x24700421","DOIUrl":"https://doi.org/10.1134/s1990519x24700421","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>This study demonstrated that silencing of HIF1A using esiRNA (endoribonuclease prepared small interfering RNA) could inhibit breast cancer cells proliferation, increase the apoptosis rate, and inhibit cell invasion and migration. These results suggested that HIF1A gene may involve in the occurrence and development of triple negative breast cancer phenotype. Hypoxia is an important hallmark of most solid tumors. In breast cancer, hypoxia is more evident in TNBC (triple negative breast cancer) than in other breast cancer subtypes. Intratumoral analysis in TNBC showed that tumor areas with high levels of hypoxia, indicated higher expression of HIF1A. Here we investigated the effect of silencing <i>HIF1A</i> on the biological function of triple negative breast cancer. Reverse transcription-polymerase chain reaction and western blot were used to detect <i>HIF1A</i> expression in different breast cancer cells and patient tissues. esiRNA was used to silence <i>HIF1A</i> in MDA-MB-231 cells. Then, BrdU assay was performed to study cell proliferation. Additionally, cell apoptosis after HIF1A knockdown was measured by Annexin V/propidium iodide staining followed by Caspase assay, and cell migration and invasion was detected by trans-well assay. IHC analysis suggested a strong overexpression of HIF1A in TNBC tissues. Silencing suggested that HIF1A significantly promotes cell proliferation and migration, resulting in metastasis and downregulates apoptosis. MDA-MB-231 cells in which HIF1A expression was inhibited were less invasive, with reduced resistance to hypoxia, impaired migration, increased apoptosis and reduced capacity to cause metastasis. HIF1A may be a dominant factor driving the metastatic progression and apoptosis of triple negative breast cancer and can be a potent therapeutic target for the disease.</p>","PeriodicalId":9705,"journal":{"name":"Cell and Tissue Biology","volume":"128 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141931650","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 : 2024-08-12DOI: 10.1134/s1990519x24700433
V. K. Rumyantseva, S. N. Morozkina, M. V. Uspenskaya, M. G. Petukhov
Abstract
Using the results of virtual ligand screening (VLS) of a representative set of 66834 commercially available drug-like ligands and 8400 di- and tripeptides in the central cavity of Transthyretin (TTR) amyloid fibrils, it has been shown that despite the great chemical diversity, among commercially available drug-like organic compounds and ultrashort peptides (USPs), only 7 USPs are able to bind in the central cavity of TTR amyloid fibrils, thus preventing the growth of amyloid fibrils. The results of VLS also show that the relaxation of ligand steric strains in the obtained complexes not only significantly improves docking scores but also radically (>50%) changes the main result of VLS, the molecular composition of 1% of the best ligands. Thus, the relaxation of steric strains after VLS can more than double the effectiveness of VLS in the development of new drugs.
{"title":"Relaxation of Steric Strains of TTR-Type Amyloid Fibril Inhibitors Radically Changes the Results of Their Virtual Screening","authors":"V. K. Rumyantseva, S. N. Morozkina, M. V. Uspenskaya, M. G. Petukhov","doi":"10.1134/s1990519x24700433","DOIUrl":"https://doi.org/10.1134/s1990519x24700433","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>Using the results of virtual ligand screening (VLS) of a representative set of 66834 commercially available drug-like ligands and 8400 di- and tripeptides in the central cavity of Transthyretin (TTR) amyloid fibrils, it has been shown that despite the great chemical diversity, among commercially available drug-like organic compounds and ultrashort peptides (USPs), only 7 USPs are able to bind in the central cavity of TTR amyloid fibrils, thus preventing the growth of amyloid fibrils. The results of VLS also show that the relaxation of ligand steric strains in the obtained complexes not only significantly improves docking scores but also radically (>50%) changes the main result of VLS, the molecular composition of 1% of the best ligands. Thus, the relaxation of steric strains after VLS can more than double the effectiveness of VLS in the development of new drugs.</p>","PeriodicalId":9705,"journal":{"name":"Cell and Tissue Biology","volume":"50 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141931649","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 : 2024-08-12DOI: 10.1134/s1990519x24700445
E. B. Menshchikova, R. A. Knyazev, N. V. Trifonova, N. A. Deeva, A. R. Kolpakov, L.P. Romakh, N. V. Kandalintseva
Abstract
The antitumor antibiotic doxorubicin, a member of a large group of anthracyclines, is widely and quite effectively used to treat patients with malignant neoplasms. However, a serious side effect of this drug is cardiotoxicity, largely due to the anthracycline’s ability to induce oxidative stress. The purpose of this study was to study the effect of TS-13, a synthetic phenolic antioxidant and activator of the redox-sensitive signaling system of the antioxidant-responsive element Keap1/Nrf2/ARE, on the functional parameters of the isolated rat heart after a course of doxorubicin administration. Male Wistar rats (n = 24) were divided into three groups: control (n = 10); a “doxorubin” group (n = 7), which received three weekly intraperitoneal injections of doxorubicin solution at a cumulative dose of 15 mg/kg; and a “doxorubicin + TS-13” group (n = 7) (doxorubicin was administered according to a similar scheme; TS-13 solution, with drinking water). On the 21st day after the start of the experiment, the presence of a cardioprotective effect of TS-13 was assessed ex vivo using a Langendorff model of the isolated heart. As parameters of myocardial functional activity, coronary flow, heart rate, and pressure in the left ventricle (myocardial contractility) were recorded; the integral indicator of myocardial contractility (performance) was calculated as the product of heart rate and pressure in the left ventricle. The general toxic effect of doxorubicin manifested itself in the form of a significant decrease in the body weight of animals (by 21%), the administration of TS-13 reduced the cachectic effect of the cytostatic. Doxorubicin worsened cardiac function in all the studied parameters (coronary flow, heart rate, myocardial contractility and integral contractility index); the effect persisted throughout the entire period of perfusion (40 min). Animals that received TS-13 per os along with intraperitoneal injections of doxorubicin lost less weight, and the functional activity of isolated hearts significantly improved: coronary flow, pressure in the left ventricle, and performance increased. We have previously shown that the administration of TS-13 not only does not cancel, but even potentiates, the antitumor activity of doxorubicin. The results obtained indicate the prospects of using TS-13 as an adjuvant therapy for malignant neoplasms, enhancing the antineoplastic effect of the cytostatic and neutralizing its side effects, including cardiotoxicity.
{"title":"Synthetic Antioxidant TS-13 Reduces the Cardiotoxicity of Doxorubicin","authors":"E. B. Menshchikova, R. A. Knyazev, N. V. Trifonova, N. A. Deeva, A. R. Kolpakov, L.P. Romakh, N. V. Kandalintseva","doi":"10.1134/s1990519x24700445","DOIUrl":"https://doi.org/10.1134/s1990519x24700445","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>The antitumor antibiotic doxorubicin, a member of a large group of anthracyclines, is widely and quite effectively used to treat patients with malignant neoplasms. However, a serious side effect of this drug is cardiotoxicity, largely due to the anthracycline’s ability to induce oxidative stress. The purpose of this study was to study the effect of TS-13, a synthetic phenolic antioxidant and activator of the redox-sensitive signaling system of the antioxidant-responsive element Keap1/Nrf2/ARE, on the functional parameters of the isolated rat heart after a course of doxorubicin administration. Male Wistar rats (<i>n</i> = 24) were divided into three groups: control (<i>n</i> = 10); a “doxorubin” group (<i>n</i> = 7), which received three weekly intraperitoneal injections of doxorubicin solution at a cumulative dose of 15 mg/kg; and a “doxorubicin + TS-13” group (<i>n</i> = 7) (doxorubicin was administered according to a similar scheme; TS-13 solution, with drinking water). On the 21st day after the start of the experiment, the presence of a cardioprotective effect of TS-13 was assessed ex vivo using a Langendorff model of the isolated heart. As parameters of myocardial functional activity, coronary flow, heart rate, and pressure in the left ventricle (myocardial contractility) were recorded; the integral indicator of myocardial contractility (performance) was calculated as the product of heart rate and pressure in the left ventricle. The general toxic effect of doxorubicin manifested itself in the form of a significant decrease in the body weight of animals (by 21%), the administration of TS-13 reduced the cachectic effect of the cytostatic. Doxorubicin worsened cardiac function in all the studied parameters (coronary flow, heart rate, myocardial contractility and integral contractility index); the effect persisted throughout the entire period of perfusion (40 min). Animals that received TS-13 per os along with intraperitoneal injections of doxorubicin lost less weight, and the functional activity of isolated hearts significantly improved: coronary flow, pressure in the left ventricle, and performance increased. We have previously shown that the administration of TS-13 not only does not cancel, but even potentiates, the antitumor activity of doxorubicin. The results obtained indicate the prospects of using TS-13 as an adjuvant therapy for malignant neoplasms, enhancing the antineoplastic effect of the cytostatic and neutralizing its side effects, including cardiotoxicity.</p>","PeriodicalId":9705,"journal":{"name":"Cell and Tissue Biology","volume":"192 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141931624","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 : 2024-08-12DOI: 10.1134/s1990519x2470041x
N. A. Obanina, N. P. Bgatova, I. D. Ivanov
Abstract
Tumor-produced pro-inflammatory cytokines and toxic substances passing through the blood-brain barrier can cause disturbances of brain homeostasis and neuronal damage. Correction of brain homeostasis under peripheral tumor growth conditions is an important task. In the present study, a cytokine expression in the brain and ultrastructural features of the prefrontal cortex pyramidal neurons in animals with skin melanoma and after lithium carbonate treatment were detected by PCR analysis, transmission electron microscopy, and immunohistochemical staining. The low expression levels of the neurotrophic factor BDNF and an increase expression of colony-stimulating factors G-CSF and GM-CSF were noted in animals with skin melanoma. At the same time, as welling of mitochondria, inner mitochondrial membrane damage and dilated endoplasmic reticulum were revealed in the prefrontal cortex neurons. Furthermore, the predominance of autophagosomes was revealed under peripheral tumor growth conditions. Lithium carbonate administration had a corrective effect on the cytokine expression in the brain and the ultrastructure of the prefrontal cortex neurons in animals with skin melanoma.
{"title":"Neurons Structure and Cytokine Expression after Lithium Carbonate Treatment on Melanoma Mice Model","authors":"N. A. Obanina, N. P. Bgatova, I. D. Ivanov","doi":"10.1134/s1990519x2470041x","DOIUrl":"https://doi.org/10.1134/s1990519x2470041x","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>Tumor-produced pro-inflammatory cytokines and toxic substances passing through the blood-brain barrier can cause disturbances of brain homeostasis and neuronal damage. Correction of brain homeostasis under peripheral tumor growth conditions is an important task. In the present study, a cytokine expression in the brain and ultrastructural features of the prefrontal cortex pyramidal neurons in animals with skin melanoma and after lithium carbonate treatment were detected by PCR analysis, transmission electron microscopy, and immunohistochemical staining. The low expression levels of the neurotrophic factor BDNF and an increase expression of colony-stimulating factors G-CSF and GM-CSF were noted in animals with skin melanoma. At the same time, as welling of mitochondria, inner mitochondrial membrane damage and dilated endoplasmic reticulum were revealed in the prefrontal cortex neurons. Furthermore, the predominance of autophagosomes was revealed under peripheral tumor growth conditions. Lithium carbonate administration had a corrective effect on the cytokine expression in the brain and the ultrastructure of the prefrontal cortex neurons in animals with skin melanoma.</p>","PeriodicalId":9705,"journal":{"name":"Cell and Tissue Biology","volume":"44 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141931623","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 : 2024-08-06DOI: 10.1134/s1990519x24700299
A. G. Goncharov, V. V. Shupletsova, N. D. Gazatova, O. B. Melashchenko, K. A. Yurova, L. S. Litvinova
Abstract
The review analyzes the results of modern high-tech research on the use of umbilical cord blood serum (plasma) as an additive to culture media for growing cell cultures. Since culture media are a key factor in cell culture, this review examines the composition and properties of the main culture media used in cell biology and regenerative medicine. The authors paid special attention to growth factors; the functional characteristics of the main families of these polypeptides are described (fibroblast growth factors, epidermal growth factors, transforming growth factors, growth differentiation, epidermal growth factors, endothelial cell growth factors, hematopoietic growth factors, etc.). It has been noted that one of the promising sources of growth factors is umbilical cord blood serum (plasma). The review presents the main technologies for obtaining umbilical cord blood, as well as systematizing studies reflecting the content of growth factors, cytokines, exosomes, and mRNA in umbilical cord blood; experimental data on the use of umbilical cord blood serum as an additive to culture media for growing various cultures of animal cells are described. Human cord blood serum, compared to animal sources, is an affordable, safe product containing high levels of bioactive molecules. For its widespread introduction as an additive to culture media, it is necessary to develop standards for the production and testing of this product.
{"title":"Cord Blood as a Trophic-Growth Additive for Culture Work","authors":"A. G. Goncharov, V. V. Shupletsova, N. D. Gazatova, O. B. Melashchenko, K. A. Yurova, L. S. Litvinova","doi":"10.1134/s1990519x24700299","DOIUrl":"https://doi.org/10.1134/s1990519x24700299","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>The review analyzes the results of modern high-tech research on the use of umbilical cord blood serum (plasma) as an additive to culture media for growing cell cultures. Since culture media are a key factor in cell culture, this review examines the composition and properties of the main culture media used in cell biology and regenerative medicine. The authors paid special attention to growth factors; the functional characteristics of the main families of these polypeptides are described (fibroblast growth factors, epidermal growth factors, transforming growth factors, growth differentiation, epidermal growth factors, endothelial cell growth factors, hematopoietic growth factors, etc.). It has been noted that one of the promising sources of growth factors is umbilical cord blood serum (plasma). The review presents the main technologies for obtaining umbilical cord blood, as well as systematizing studies reflecting the content of growth factors, cytokines, exosomes, and mRNA in umbilical cord blood; experimental data on the use of umbilical cord blood serum as an additive to culture media for growing various cultures of animal cells are described. Human cord blood serum, compared to animal sources, is an affordable, safe product containing high levels of bioactive molecules. For its widespread introduction as an additive to culture media, it is necessary to develop standards for the production and testing of this product.</p>","PeriodicalId":9705,"journal":{"name":"Cell and Tissue Biology","volume":"73 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141931652","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}
The sequential culture expansion has been shown to affect the favorable characteristics of human mesenchymal stem/stromal cells (hMSCs), including their immunomodulatory potential. Hereby, we have investigated the immunomodulatory potential of Yazd human testicular Cells (YhTCs) regarding gene expression profile assessment during their lifespan. Morphological examination, immunostaining of vimentin and fibronectin markers, and flow cytometry analysis were used to confirm cultured cells’ MSC properties. Quantitative RT-PCR was used to investigate the target genes, COX-1, TGF-β, IL-6, VCAM-1, and FAS, at different passage numbers (P3, P6, and P9). Our findings confirmed the MSC properties of YhTCs over the lifespan. The results of COX-1 and IL-6 indicated significantly higher expression levels at the late passages (p‑value = 0.03 and p-value = 0.01, respectively). Despite the similar expression pattern, VCAM-1 expression altered significantly at different passages (p-value = 0.0006), but the changes in TGF-β expression levels were non-significant across the lifespan. The YhTCs maintained the main characteristics of MSCs over the sequential culture expansion. Although some of the target genes in the study were sensitive to the culture expansion in the unstimulated YhTCs (IL-6, COX-1, and VCAM1), it seems that FAS and TGF-β need external stimuli or intracellular signals to be induced in YhTCs.
{"title":"Immunomodulatory Potential of Human Testicular Tissue-Derived Mesenchymal Stem/Stromal Cells over the Lifespan","authors":"Maryam Hassan Nasab, Mahmood Dehghani-Ashkezari, Fateme Montazeri, Ehsan Farashahi-Yazd, Fatemeh Hajizadeh-Tafti, Jalal Golzadeh, Seyed Mehdi Hoseini, Behrouz Aflatoonian","doi":"10.1134/s1990519x24700329","DOIUrl":"https://doi.org/10.1134/s1990519x24700329","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>The sequential culture expansion has been shown to affect the favorable characteristics of human mesenchymal stem/stromal cells (hMSCs), including their immunomodulatory potential. Hereby, we have investigated the immunomodulatory potential of Yazd human testicular Cells (YhTCs) regarding gene expression profile assessment during their lifespan. Morphological examination, immunostaining of vimentin and fibronectin markers, and flow cytometry analysis were used to confirm cultured cells’ MSC properties. Quantitative RT-PCR was used to investigate the target genes, <i>COX-1</i>, <i>TGF-</i>β, <i>IL-6</i>, <i>VCAM-1,</i> and <i>FAS</i>, at different passage numbers (P3, P6, and P9). Our findings confirmed the MSC properties of YhTCs over the lifespan. The results of <i>COX-1</i> and <i>IL-6</i> indicated significantly higher expression levels at the late passages (<i>p</i>‑value = 0.03 and <i>p</i>-value = 0.01, respectively). Despite the similar expression pattern, <i>VCAM-1</i> expression altered significantly at different passages (<i>p</i>-value = 0.0006), but the changes in <i>TGF-</i>β expression levels were non-significant across the lifespan. The YhTCs maintained the main characteristics of MSCs over the sequential culture expansion. Although some of the target genes in the study were sensitive to the culture expansion in the unstimulated YhTCs (<i>IL-6</i>, <i>COX-1,</i> and <i>VCAM1</i>), it seems that <i>FAS</i> and <i>TGF-</i>β need external stimuli or intracellular signals to be induced in YhTCs.</p>","PeriodicalId":9705,"journal":{"name":"Cell and Tissue Biology","volume":"22 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141931655","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 : 2024-08-06DOI: 10.1134/s1990519x24700408
L. M. Polyakov, D. V. Sumenkova, M. V. Kotova, N. V. Trifonova, R. A. Knyazev
Abstract
The purpose of this work was to study the possibility of using apolipoprotein A-I (apo A-I) as a transport variant form of green fluorescent protein (GFP) gene in rat hepatocytes. Cultured rat hepatocytes were used as a model. The conjugate of apo A-I with fluorescein isothiocyanate (FITC) was prepared by incubation of the apo A-I protein with FITC in carbonate buffer, pH 9.5, at a ratio of 1.5 μg of FITC per 1 mg of the protein. Plasmids for pE-GAG transfection with an inserted GFP gene were enriched in the promoter region with cis-elements of CC(GCC)3-5 type to enhance complex formation with apo A-I. An inverted fluorescence microscope was used to visually analyze cell fluorescence. The work presents evidence of penetration of FITC-labeled apo A-I into the cytoplasm and nuclei of rat hepatocytes through receptor-mediated endocytosis. Based on this evidence, it was attempted to use apo A-I as a an agent of targeted delivery of plasmid DNA with an inserted GFP gene into a cell. According to the results of fluorescence microscopy, using apo A-I as a transfection agent for plasmid DNA resulted in accumulation of GFP protein in the cytoplasm of hepatocytes. Expression of the GFP gene and accumulation of fluorescent protein were not observed in the absence of apo A-I. The results obtained may indicate the delivery of GFP gene to the nuclear apparatus of the cell, its expression and synthesis of GFP protein.
{"title":"Apolipoprotein A-I as a Transport Alternative of Green Fluorescent Protein GFP Gene in Rat Hepatocytes","authors":"L. M. Polyakov, D. V. Sumenkova, M. V. Kotova, N. V. Trifonova, R. A. Knyazev","doi":"10.1134/s1990519x24700408","DOIUrl":"https://doi.org/10.1134/s1990519x24700408","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>The purpose of this work was to study the possibility of using apolipoprotein A-I (apo A-I) as a transport variant form of green fluorescent protein (GFP) gene in rat hepatocytes. Cultured rat hepatocytes were used as a model. The conjugate of apo A-I with fluorescein isothiocyanate (FITC) was prepared by incubation of the apo A-I protein with FITC in carbonate buffer, pH 9.5, at a ratio of 1.5 μg of FITC per 1 mg of the protein. Plasmids for pE-GAG transfection with an inserted GFP gene were enriched in the promoter region with <i>cis</i>-elements of CC(GCC)3-5 type to enhance complex formation with apo A-I. An inverted fluorescence microscope was used to visually analyze cell fluorescence. The work presents evidence of penetration of FITC-labeled apo A-I into the cytoplasm and nuclei of rat hepatocytes through receptor-mediated endocytosis. Based on this evidence, it was attempted to use apo A-I as a an agent of targeted delivery of plasmid DNA with an inserted GFP gene into a cell. According to the results of fluorescence microscopy, using apo A-I as a transfection agent for plasmid DNA resulted in accumulation of GFP protein in the cytoplasm of hepatocytes. Expression of the GFP gene and accumulation of fluorescent protein were not observed in the absence of apo A-I. The results obtained may indicate the delivery of GFP gene to the nuclear apparatus of the cell, its expression and synthesis of GFP protein.</p>","PeriodicalId":9705,"journal":{"name":"Cell and Tissue Biology","volume":"58 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141931708","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 : 2024-08-06DOI: 10.1134/s1990519x24700354
S. A. Vladimirova, B. A. Margulis, I. V. Guzhova, A. D. Nicotina
Abstract
The occurrence of intolerable side effects in patients undergoing chemotherapy continues to be a key clinical obstacle. In this regard, the search for tumor-specific therapy that does not have a toxic effect on healthy tissue remains an urgent task. It is known that heat-shock protein factor HSF1 is an important marker of cancer progression, and the products of its transcriptional activity allow tumor cells to successfully avoid the negative effects of antitumor therapy. Thereby, the use of drugs that inhibit HSF1 activity is a promising strategy. In this research, we found that the HSF1 activity inhibitor from the CL-43 cardenolide group exhibited a cytoprotective effect on primary nontransformed dermal fibroblast cells (DF-2) and made them less sensitive to etoposide, while in DLD1 tumor cells, on the contrary, we observed an increase in this sensitivity. In addition, we found that CL-43 affected the intranuclear transport of active HSF1, as well as increasing its activity and, accordingly, the synthesis of HSP70 in human fibroblasts, while CL-43 suppressed this activity in a dose-dependent manner in tumor cells. Our results indicate the high therapeutic potential of CL-43 and its uniqueness as a tumor-specific compound.
{"title":"The Effect of HSF1 Activity Inhibitor of the Cardenolide Group (CL-43) on Tumor and Nontransformed Cells","authors":"S. A. Vladimirova, B. A. Margulis, I. V. Guzhova, A. D. Nicotina","doi":"10.1134/s1990519x24700354","DOIUrl":"https://doi.org/10.1134/s1990519x24700354","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>The occurrence of intolerable side effects in patients undergoing chemotherapy continues to be a key clinical obstacle. In this regard, the search for tumor-specific therapy that does not have a toxic effect on healthy tissue remains an urgent task. It is known that heat-shock protein factor HSF1 is an important marker of cancer progression, and the products of its transcriptional activity allow tumor cells to successfully avoid the negative effects of antitumor therapy. Thereby, the use of drugs that inhibit HSF1 activity is a promising strategy. In this research, we found that the HSF1 activity inhibitor from the CL-43 cardenolide group exhibited a cytoprotective effect on primary nontransformed dermal fibroblast cells (DF-2) and made them less sensitive to etoposide, while in DLD1 tumor cells, on the contrary, we observed an increase in this sensitivity. In addition, we found that CL-43 affected the intranuclear transport of active HSF1, as well as increasing its activity and, accordingly, the synthesis of HSP70 in human fibroblasts, while CL-43 suppressed this activity in a dose-dependent manner in tumor cells. Our results indicate the high therapeutic potential of CL-43 and its uniqueness as a tumor-specific compound.</p>","PeriodicalId":9705,"journal":{"name":"Cell and Tissue Biology","volume":"192 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141931659","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 : 2024-08-06DOI: 10.1134/s1990519x24700391
E. V. Bogdanenko, L. A. Sergievich, A. V. Karnaukhov, N. A. Karnaukhova, I. A. Lizunova
Abstract
In сlinical practice, allogeneiс bone marrow transplantation (BMT) is often a cause of graft-vs.-host disease (GvHD). GvHD is explained by the fact that T-lymphocytes, which are administered simultaneously with hematopoietic cells during transplantation and form and mature anew in the timus of the recipient from donor progenitor cells thereafter, recognize and attack the cells of the host. However, a complete explanation of the phenomenon of the GvHD does not exist, and chimerization of the recipient’s organism as a possible cause of damage of its organs is not taken into account. Therefore, the aim of this work consisted in modeling of allogeneic transplantation of the whole bone marrow (experiment) and comparing its results with syngeneic transplantation (control) basing on an investigation of engraftment of cells of donor origin in the main GvHD target organs. The bone marrow donors were Tg(ACTB-EGFP)1Osb/J mice carrying a green fluorescent protein gene (EGFP), and the recipients were animals of CBA and C57BL/6 inbred strains of an age of 2–10 months. One day before BMT (1.5 × 107 cells per mouse), all recipients were irradiated at a dose of 6.5 Gy (LD 50/30). After 1, 3, 5, 7, 11, 14, 21, 28, 35, and 55 days, the development of chimerism in the liver, skin, and colon of animals was examined using a fluorescent microscope. Already after 1 day, single fibroblast-like donor cells were found in the colon; after 7 days, in the skin and liver. From 14 to 28 days after BMT, donor cells formed mainly stroma in the liver, as well as fibroblasts and keratinocytes in the skin, and in the colon they substituted the cells of the villi, stroma, and parenchyma of Peyer’s patches that died after irradiation. Unlike control, in the experimental groups, GFP+ giant fibroblasts about 30 μm in length were found in the stroma of the liver, as well as in the skin and in the colon; in the liver there were many GFP+ bulkheads and fibroblast-like Ito’s cells of very intricate configuration. From 35 to 55 days after allogeneic BMT, cells of the donor origin in the liver and in the villi of the colon began to destroy, the villi became overgrown with GFP+ connective tissue cells and warped, the wall of the colon became thin, and the skin was fully replaced with a new one (these things were never observed in the control groups). We propose the hypothesis that, alongside GvHD traits like thinning of the colon wall and a large number of roundish GFP+ cells on the inner surface of the skin, other signs of the studied after allogeneic BMT organs suggest that the cells of the organs that are formed from mesenchymal stem cells of the whole bone marrow become targets for the recipient’s T cells, i.e., suggest the existence of a host-vs.-graft (HVG) reaction. The obvious manifestation of immune reactions after BMT directly coincides with the term of massive engraftment of the studied organs with cells of donor origin
{"title":"A Study of the Relationship of the Dynamics of Development and Characteristics of Chimerism with Manifestations of Graft-vs.-Host Disease in the Organs of Mice after Allogeneic Transplantation of Whole Bone Marrow","authors":"E. V. Bogdanenko, L. A. Sergievich, A. V. Karnaukhov, N. A. Karnaukhova, I. A. Lizunova","doi":"10.1134/s1990519x24700391","DOIUrl":"https://doi.org/10.1134/s1990519x24700391","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>In сlinical practice, allogeneiс bone marrow transplantation (BMT) is often a cause of graft-vs.-host disease (GvHD). GvHD is explained by the fact that T-lymphocytes, which are administered simultaneously with hematopoietic cells during transplantation and form and mature anew in the timus of the recipient from donor progenitor cells thereafter, recognize and attack the cells of the host. However, a complete explanation of the phenomenon of the GvHD does not exist, and chimerization of the recipient’s organism as a possible cause of damage of its organs is not taken into account. Therefore, the aim of this work consisted in modeling of allogeneic transplantation of the whole bone marrow (experiment) and comparing its results with syngeneic transplantation (control) basing on an investigation of engraftment of cells of donor origin in the main GvHD target organs. The bone marrow donors were Tg(ACTB-EGFP)1Osb/J mice carrying a green fluorescent protein gene (EGFP), and the recipients were animals of CBA and C57BL/6 inbred strains of an age of 2–10 months. One day before BMT (1.5 × 10<sup>7</sup> cells per mouse), all recipients were irradiated at a dose of 6.5 Gy (LD 50/30). After 1, 3, 5, 7, 11, 14, 21, 28, 35, and 55 days, the development of chimerism in the liver, skin, and colon of animals was examined using a fluorescent microscope. Already after 1 day, single fibroblast-like donor cells were found in the colon; after 7 days, in the skin and liver. From 14 to 28 days after BMT, donor cells formed mainly stroma in the liver, as well as fibroblasts and keratinocytes in the skin, and in the colon they substituted the cells of the villi, stroma, and parenchyma of Peyer’s patches that died after irradiation. Unlike control, in the experimental groups, GFP+ giant fibroblasts about 30 μm in length were found in the stroma of the liver, as well as in the skin and in the colon; in the liver there were many GFP+ bulkheads and fibroblast-like Ito’s cells of very intricate configuration. From 35 to 55 days after allogeneic BMT, cells of the donor origin in the liver and in the villi of the colon began to destroy, the villi became overgrown with GFP+ connective tissue cells and warped, the wall of the colon became thin, and the skin was fully replaced with a new one (these things were never observed in the control groups). We propose the hypothesis that, alongside GvHD traits like thinning of the colon wall and a large number of roundish GFP+ cells on the inner surface of the skin, other signs of the studied after allogeneic BMT organs suggest that the cells of the organs that are formed from mesenchymal stem cells of the whole bone marrow become targets for the recipient’s T cells, i.e., suggest the existence of a host-vs.-graft (HVG) reaction. The obvious manifestation of immune reactions after BMT directly coincides with the term of massive engraftment of the studied organs with cells of donor origin","PeriodicalId":9705,"journal":{"name":"Cell and Tissue Biology","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141931662","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}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号