AIM: To make drug sera of Salvia miltiorrhiza and Yigankang, both of which are Chinese herbs that activate bleeding and eliminate stasis, in normal rats and those with liver fibrosis, respectively. To investigate and ...AIM: To make drug sera of Salvia miltiorrhiza and Yigankang, both of which are Chinese herbs that activate bleeding and eliminate stasis, in normal rats and those with liver fibrosis, respectively. To investigate and compare the effects of the two different drug sera on the proliferation and activation of hepatic stellate cells (HSCs).METHODS: Some rats were induced with liver fibrosis:40% carbon tetrachloride (CCl4) subcutaneous injection,twice a week for 9 wk. Salvia miltiorrhiza, Yigankang,colchicines and normal saline were administered into the stomachs of normal rats and those with liver fibrosis.Drug sera were extracted 5 d later. HSCs in vitro were cultivated in different drug sera for 24 h. The rates of proliferation and expression of α-smooth muscle actin (α-SMA) were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and immunocytochemistry stain, respectively.RESULTS: The drug sera from normal and liver fibrotic rats could be used to cultivate HSCs and to observe the effects of the corresponding components of herbs on HSCs. Salvia miltiorrhiza and Yigankang had better inhibitory effects on HSCs than colchicines (MTT: normal drug serum: Salvia miltiorrhiza 0.42±0.08, Yigankang 0.32±0.10 vs colchicines 0.45±0.12 pathological drug serum: Salvia miltiorrhiza 0.33±0.02, Yigankang 0.26±0.01vs colchicines 0.41±0.09. P＜0.05). The drug sera of Salvia miltiorrhiza, Yigankang from liver fibrotic rats had a stronger inhibitory effect than the same ones from normal rats (MTT: Salvia miltiorrhiza: normal drug serum 0.42±0.08 vs pathological drug serum 0.33±0.02. Yigankang: normal drug serum 0.32±0.10 vs pathological drug serum 0.26±0.01.P＜0.05).CONCLUSION: Salvia miltiorrhiza and Yigankang could inhibit the expression of α-SMA and the proliferation of HSCs. The drug sera from normal and liver fibrotic rats had different effects on HSCs, probably due to different metabolic processes, effective components and different quantities of drug contents in drug sera from rats with different states of liver.展开更多
Investigating the function of combining induced rat monocytes-derived bone marrow-haemopoietic stem cell (rat BM-HSCs) with LPS and rat bone marrow-mesenchymal stem cell (rat BM-MSCs) was to analyze the acceleration o...Investigating the function of combining induced rat monocytes-derived bone marrow-haemopoietic stem cell (rat BM-HSCs) with LPS and rat bone marrow-mesenchymal stem cell (rat BM-MSCs) was to analyze the acceleration of homing process mechanism in injured pancreas. Mononucleated stem cells were isolated from aspirated whole rat BM using ficoll and cultured in α-MEM complete growth medium in 10 cm petridish. After two days, adherent cells after washing twice in petridish were added α-MEM growth medium and then mesenchymal cells were characterized using CD105 marker in third passage and labeled PKH26. Then haemopoietic stem cells (HSCs) were isolated with magnetic beads CD34+ and differentiated in vitro, and then induced monocytes with LPS. Animal experiment used 28 male Wistar rats, and divided them into 4 groups. After transplantation combined, both cells between monocyte derived HSc (mHSCs) and rat BM-MSC were analyzed expression of pair box gen 4 (Pax4), pancreatic and duodenal homeobox (Pdx1), C-peptide using immunohistochemistry, then secretion of insulin and C-peptide analyzed using indirect ELISA. Results showed that the expressions of Pax4, Pdx1, C-peptide found in the surface membrane cell of pancreatic cell, and secreted C-peptide and insulin were shown significant (P < 0.05) in transplanted group 2, 3 and 4, but in group 3 were transplanted with combined cells more dominant than non-combined cells. Conclusions suggested that combining of induced monocytes-derived HSCs and rat BM-MSCs has accelerated homing MSCs into injured pancreatic tissue.展开更多
Hepatic stellate cell （HSC） activation is an essential event during liver fibrogene- sis. Phosphatase and tension homolog deleted on chromosome 10 （PTEN）, a tumor suppressor, is a negative regulator of this proces...Hepatic stellate cell （HSC） activation is an essential event during liver fibrogene- sis. Phosphatase and tension homolog deleted on chromosome 10 （PTEN）, a tumor suppressor, is a negative regulator of this process. PTEN promoter hypermethylation is a major epigenetic si- lencing mechanism in tumors. The present study aimed to investigate whether PTEN promoter methylation was involved in HSCs activation and liver fibrosis, we observed that hypermethyla- tion of PTEN gene was responsible for the decrease of PTEN expression during HSCs展开更多
A study recently published in Nature reported a single-cell transcriptome map of human hematopoietic stem cells(HSCs)and a gene expression signature that distinguishes nascent HSCs from non-HSCs during prenatal develo...A study recently published in Nature reported a single-cell transcriptome map of human hematopoietic stem cells(HSCs)and a gene expression signature that distinguishes nascent HSCs from non-HSCs during prenatal development.1 This transcriptome map provides an important tool for further elucidation of human HSC ontogeny and could also serve as a guide for generation of transplantable HSCs ex vivo,1 to widen the therapeutic application of HSCs.展开更多
An effective therapeutic regimen for hepatic fibrosis requires a deep understanding of the pathogenesis mechanism.Hepatic fibrosis is characterized by activated hepatic stellate cells(aHSCs)with an excessive productio...An effective therapeutic regimen for hepatic fibrosis requires a deep understanding of the pathogenesis mechanism.Hepatic fibrosis is characterized by activated hepatic stellate cells(aHSCs)with an excessive production of extracellular matrix.Although promoted activation of HSCs by M2 macrophages has been demonstrated,the molecular mechanism involved remains ambiguous.Herein,we propose that the vitamin D receptor(VDR)involved in macrophage polarization may regulate the communication between macrophages and HSCs by changing the functions of exosomes.We confirm that activating the VDR can inhibit the effect of M2 macrophages on HSC activation.The exosomes derived from M2 macrophages can promote HSC activation,while stimulating VDR alters the protein profiles and reverses their roles in M2 macrophage exosomes.Smooth muscle cell-associated protein 5(SMAP-5)was found to be the key effector protein in promoting HSC activation by regulating autophagy flux.Building on these results,we show that a combined treatment of a VDR agonist and a macrophage-targeted exosomal secretion inhibitor achieves an excellent anti-hepatic fibrosis effect.In this study,we aim to elucidate the association between VDR and macrophages in HSC activation.The results contribute to our understanding of the pathogenesis mechanism of hepatic fibrosis,and provide potential therapeutic targets for its treatment.展开更多
Objective:To investigate the role and mechanism of mTOR in sorafenib-ameliorated liver fibrosis by inducing hepatic stellate cells(HSC)ferroptosis.Methods:The liver fibrosis models of C57BL/6 male mice were induced wi...Objective:To investigate the role and mechanism of mTOR in sorafenib-ameliorated liver fibrosis by inducing hepatic stellate cells(HSC)ferroptosis.Methods:The liver fibrosis models of C57BL/6 male mice were induced with carbon tetrachloride(CCl4)and randomly divided into normal group,model group,and sorafenib low(2.5 mg/kg),medium(5 mg/kg),and high(10 mg/kg)dose groups.Except for the normal group,the remaining four groups were treated with intraperitoneal injection of CCl4 for 8 weeks to establish liver fibrosis models.In addition,the corresponding concentrations of sorafenib were administered during the fourth week of modeling.Western blot was used to detect the expression of mTOR and p-mTOR protein in liver tissues.In vitro experiments,HSC-T6 cells were activated by PDGFBB and then treated with sorafenib(5μM,10μM,20μM),an mTOR inhibitor and activator.CCK8 was used to detect HSC-T6 cell viability,Western blot detected the protein expression ofα-SMA,COL1α1,GPX4,mTOR and p-mTOR.The levels of serum iron,GSH,and MDA were measured,and the intracellular ROS level was measured by 2',7'-dichlorofluorescein diacetate and dihydroethidium.Results:In vitro results showed that sorafenib significantly decreased the protein expression ofα-SMA,COL1α1,GPX4 and p-mTOR,and decreased the level of GSH and increased the content of iron,MDA and ROS in HSC-T6(P<0.05).Sorafenib(5μM,10μM,20μM)significantly inhibited the cell viability of PDGF-BB-enhanced HSCT6(P<0.05).When mTOR was inhibited,the protein expressions ofα-SMA,COL1α1 and GPX4 and the level of GSH were lower(P<0.05),and the contents of iron,MDA and ROS were further increased(P<0.05).When mTOR was activated,the results were reversed.Conclusion:Sorafenib induced ferroptosis to alleviate liver fibrosis,and inhibition of mTOR further enhanced the effect of sorafenib.展开更多
文摘AIM: To make drug sera of Salvia miltiorrhiza and Yigankang, both of which are Chinese herbs that activate bleeding and eliminate stasis, in normal rats and those with liver fibrosis, respectively. To investigate and compare the effects of the two different drug sera on the proliferation and activation of hepatic stellate cells (HSCs).METHODS: Some rats were induced with liver fibrosis:40% carbon tetrachloride (CCl4) subcutaneous injection,twice a week for 9 wk. Salvia miltiorrhiza, Yigankang,colchicines and normal saline were administered into the stomachs of normal rats and those with liver fibrosis.Drug sera were extracted 5 d later. HSCs in vitro were cultivated in different drug sera for 24 h. The rates of proliferation and expression of α-smooth muscle actin (α-SMA) were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and immunocytochemistry stain, respectively.RESULTS: The drug sera from normal and liver fibrotic rats could be used to cultivate HSCs and to observe the effects of the corresponding components of herbs on HSCs. Salvia miltiorrhiza and Yigankang had better inhibitory effects on HSCs than colchicines (MTT: normal drug serum: Salvia miltiorrhiza 0.42±0.08, Yigankang 0.32±0.10 vs colchicines 0.45±0.12 pathological drug serum: Salvia miltiorrhiza 0.33±0.02, Yigankang 0.26±0.01vs colchicines 0.41±0.09. P＜0.05). The drug sera of Salvia miltiorrhiza, Yigankang from liver fibrotic rats had a stronger inhibitory effect than the same ones from normal rats (MTT: Salvia miltiorrhiza: normal drug serum 0.42±0.08 vs pathological drug serum 0.33±0.02. Yigankang: normal drug serum 0.32±0.10 vs pathological drug serum 0.26±0.01.P＜0.05).CONCLUSION: Salvia miltiorrhiza and Yigankang could inhibit the expression of α-SMA and the proliferation of HSCs. The drug sera from normal and liver fibrotic rats had different effects on HSCs, probably due to different metabolic processes, effective components and different quantities of drug contents in drug sera from rats with different states of liver.
文摘Investigating the function of combining induced rat monocytes-derived bone marrow-haemopoietic stem cell (rat BM-HSCs) with LPS and rat bone marrow-mesenchymal stem cell (rat BM-MSCs) was to analyze the acceleration of homing process mechanism in injured pancreas. Mononucleated stem cells were isolated from aspirated whole rat BM using ficoll and cultured in α-MEM complete growth medium in 10 cm petridish. After two days, adherent cells after washing twice in petridish were added α-MEM growth medium and then mesenchymal cells were characterized using CD105 marker in third passage and labeled PKH26. Then haemopoietic stem cells (HSCs) were isolated with magnetic beads CD34+ and differentiated in vitro, and then induced monocytes with LPS. Animal experiment used 28 male Wistar rats, and divided them into 4 groups. After transplantation combined, both cells between monocyte derived HSc (mHSCs) and rat BM-MSC were analyzed expression of pair box gen 4 (Pax4), pancreatic and duodenal homeobox (Pdx1), C-peptide using immunohistochemistry, then secretion of insulin and C-peptide analyzed using indirect ELISA. Results showed that the expressions of Pax4, Pdx1, C-peptide found in the surface membrane cell of pancreatic cell, and secreted C-peptide and insulin were shown significant (P < 0.05) in transplanted group 2, 3 and 4, but in group 3 were transplanted with combined cells more dominant than non-combined cells. Conclusions suggested that combining of induced monocytes-derived HSCs and rat BM-MSCs has accelerated homing MSCs into injured pancreatic tissue.
文摘Hepatic stellate cell （HSC） activation is an essential event during liver fibrogene- sis. Phosphatase and tension homolog deleted on chromosome 10 （PTEN）, a tumor suppressor, is a negative regulator of this process. PTEN promoter hypermethylation is a major epigenetic si- lencing mechanism in tumors. The present study aimed to investigate whether PTEN promoter methylation was involved in HSCs activation and liver fibrosis, we observed that hypermethyla- tion of PTEN gene was responsible for the decrease of PTEN expression during HSCs
基金The work was supported by grants from the Forschungskommission of the Medical Faculty of the Heinrich Heine University of Düsseldorf and the Leukämie Lymphom Liga e.V.to E.Grinstein.Acknowledged is furthermore grant support from Deutsche Forschungsgemeinschaft to E.Grinstein.
文摘A study recently published in Nature reported a single-cell transcriptome map of human hematopoietic stem cells(HSCs)and a gene expression signature that distinguishes nascent HSCs from non-HSCs during prenatal development.1 This transcriptome map provides an important tool for further elucidation of human HSC ontogeny and could also serve as a guide for generation of transplantable HSCs ex vivo,1 to widen the therapeutic application of HSCs.
基金supported by the National Natural Science Foundation of China(Nos.81930099,81773664,82130102,92159304,81703585,and 81903651)the Natural Science Foundation of Jiangsu Province(Nos.BK20212011 and BK20180565)+4 种基金the Technology Innovation Project of Nucleic Acid Drug from National Center of Technology Innovation for Biopharmaceuticals(No.NCTIB2022HS01014)the“Double First-Class”University Project(No.CPU2022QZ05)the 111 Project from the Ministry of Education of China and the State Administration of Foreign Expert Affairs of China(Nos.111-2-07 and B17047)the Fundamental Research Funds for the Central Universities of China(No.2632022ZD11)the Open Project of State Key Laboratory of Natural Medicines(No.SKLNMZZ202017),China.
文摘An effective therapeutic regimen for hepatic fibrosis requires a deep understanding of the pathogenesis mechanism.Hepatic fibrosis is characterized by activated hepatic stellate cells(aHSCs)with an excessive production of extracellular matrix.Although promoted activation of HSCs by M2 macrophages has been demonstrated,the molecular mechanism involved remains ambiguous.Herein,we propose that the vitamin D receptor(VDR)involved in macrophage polarization may regulate the communication between macrophages and HSCs by changing the functions of exosomes.We confirm that activating the VDR can inhibit the effect of M2 macrophages on HSC activation.The exosomes derived from M2 macrophages can promote HSC activation,while stimulating VDR alters the protein profiles and reverses their roles in M2 macrophage exosomes.Smooth muscle cell-associated protein 5(SMAP-5)was found to be the key effector protein in promoting HSC activation by regulating autophagy flux.Building on these results,we show that a combined treatment of a VDR agonist and a macrophage-targeted exosomal secretion inhibitor achieves an excellent anti-hepatic fibrosis effect.In this study,we aim to elucidate the association between VDR and macrophages in HSC activation.The results contribute to our understanding of the pathogenesis mechanism of hepatic fibrosis,and provide potential therapeutic targets for its treatment.
文摘单钢板混凝土组合(half steel-concrete composite,HSC)板的抗冲击性能十分出色,可有效抵挡落石冲击。在符合相关规范要求的前提下,可通过设计降低HSC板在落石冲击作用下的变形,以最大限度地保护山区基础设施和人民的安全。为快速、准确地处理HSC板的变形和设计参数之间的复杂非线性关系,基于三种机器学习算法分别建立冲击作用下HSC板最大变形的预测模型,并通过有限元结果对模型进行验证;在此基础上,以落石冲击下HSC板的最大变形、自质量和造价为优化目标,采用遗传算法对某山区建筑中某HSC屋面板进行优化设计,求解HSC屋面板的厚度、连接件尺寸等设计参数的最优解集。研究结果表明,高斯过程回归(Gaussian process regression,GPR)模型对HSC屋面板变形的预测精度最高,可代替复杂耗时的有限元计算,有效提高目标方程的计算效率,且最终的优化结果给出了多种优化方案,可有效降低HSC屋面板变形,为工程设计提供参考。
基金Foundation Project of National Natural Science(81600498)。
文摘Objective:To investigate the role and mechanism of mTOR in sorafenib-ameliorated liver fibrosis by inducing hepatic stellate cells(HSC)ferroptosis.Methods:The liver fibrosis models of C57BL/6 male mice were induced with carbon tetrachloride(CCl4)and randomly divided into normal group,model group,and sorafenib low(2.5 mg/kg),medium(5 mg/kg),and high(10 mg/kg)dose groups.Except for the normal group,the remaining four groups were treated with intraperitoneal injection of CCl4 for 8 weeks to establish liver fibrosis models.In addition,the corresponding concentrations of sorafenib were administered during the fourth week of modeling.Western blot was used to detect the expression of mTOR and p-mTOR protein in liver tissues.In vitro experiments,HSC-T6 cells were activated by PDGFBB and then treated with sorafenib(5μM,10μM,20μM),an mTOR inhibitor and activator.CCK8 was used to detect HSC-T6 cell viability,Western blot detected the protein expression ofα-SMA,COL1α1,GPX4,mTOR and p-mTOR.The levels of serum iron,GSH,and MDA were measured,and the intracellular ROS level was measured by 2',7'-dichlorofluorescein diacetate and dihydroethidium.Results:In vitro results showed that sorafenib significantly decreased the protein expression ofα-SMA,COL1α1,GPX4 and p-mTOR,and decreased the level of GSH and increased the content of iron,MDA and ROS in HSC-T6(P<0.05).Sorafenib(5μM,10μM,20μM)significantly inhibited the cell viability of PDGF-BB-enhanced HSCT6(P<0.05).When mTOR was inhibited,the protein expressions ofα-SMA,COL1α1 and GPX4 and the level of GSH were lower(P<0.05),and the contents of iron,MDA and ROS were further increased(P<0.05).When mTOR was activated,the results were reversed.Conclusion:Sorafenib induced ferroptosis to alleviate liver fibrosis,and inhibition of mTOR further enhanced the effect of sorafenib.
