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
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
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
目的:研究五味子甲素对人肝星状细胞系LX-2细胞的增殖、纤维化指标及凋亡相关蛋白的影响。方法:采用不同浓度的五味子甲素与肝星状细胞LX-2共孵育,MTT法检测五味子甲素对细胞的抑制作用,RT-PCR法和Western blot法测定纤维化指标I型胶原...目的:研究五味子甲素对人肝星状细胞系LX-2细胞的增殖、纤维化指标及凋亡相关蛋白的影响。方法:采用不同浓度的五味子甲素与肝星状细胞LX-2共孵育,MTT法检测五味子甲素对细胞的抑制作用,RT-PCR法和Western blot法测定纤维化指标I型胶原α亚基1(collagen type I alpha 1,collagen I-1)和α-平滑肌动蛋白(α-smooth muscle actin,α-SMA)的表达量,并检测五味子甲素对凋亡相关蛋白Bcl-2和Bax表达量的影响。结果:五味子甲素在不同浓度时可抑制人肝星状细胞系LX-2细胞的增殖,可降低collagenⅠ和α-SMA的mRNA水平和蛋白表达量(P<0.05),并能降低Bcl-2的mRNA水平和蛋白表达量以及Bcl-2/Bax的比值(P<0.05)。结论:五味子甲素可能通过抑制细胞增殖、降低纤维化指标的表达,从而发挥抗肝纤维化的作用。展开更多
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.
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.
文摘目的:研究五味子甲素对人肝星状细胞系LX-2细胞的增殖、纤维化指标及凋亡相关蛋白的影响。方法:采用不同浓度的五味子甲素与肝星状细胞LX-2共孵育,MTT法检测五味子甲素对细胞的抑制作用,RT-PCR法和Western blot法测定纤维化指标I型胶原α亚基1(collagen type I alpha 1,collagen I-1)和α-平滑肌动蛋白(α-smooth muscle actin,α-SMA)的表达量,并检测五味子甲素对凋亡相关蛋白Bcl-2和Bax表达量的影响。结果:五味子甲素在不同浓度时可抑制人肝星状细胞系LX-2细胞的增殖,可降低collagenⅠ和α-SMA的mRNA水平和蛋白表达量(P<0.05),并能降低Bcl-2的mRNA水平和蛋白表达量以及Bcl-2/Bax的比值(P<0.05)。结论:五味子甲素可能通过抑制细胞增殖、降低纤维化指标的表达,从而发挥抗肝纤维化的作用。
