中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2015年
23期
3639-3643
,共5页
干细胞%骨髓干细胞%肝星状细胞%骨髓间充质干细胞%肝细胞生长因子%肝细胞生长因子激活因子%细胞凋亡%机制
榦細胞%骨髓榦細胞%肝星狀細胞%骨髓間充質榦細胞%肝細胞生長因子%肝細胞生長因子激活因子%細胞凋亡%機製
간세포%골수간세포%간성상세포%골수간충질간세포%간세포생장인자%간세포생장인자격활인자%세포조망%궤제
Hepatocytes%Apoptosis%Proto-Oncogene Protein c-met%rho-Associated Kinases
背景：控制肝星状细胞的激活和增殖是抗肝纤维化研究的重点，人类或鼠的骨髓间充质干细胞可通过旁分泌的肝细胞生长因子诱导肝星状细胞凋亡。目的：探讨骨髓间充质干细胞在肝星状细胞凋亡中的参与机制。方法：构建共培养体系，于半透膜上下层在细胞培养板上对肝星状细胞和骨髓间充质干细胞进行接种和培养，单纯肝星状细胞组设为空白对照组，共培养的肝星状细胞和骨髓间充质干细胞组为共培养组，以3 mg/L c-Met阻滞剂干预的肝星状细胞和骨髓间充质干细胞为c-Met阻滞剂组，以3 mg/L RhoA阻滞剂干预的肝星状细胞和骨髓间充质干细胞为RhoA阻滞剂组。结果与结论：c-Met阻滞剂质量浓度为3.0 mg/L，RhoA阻滞剂浓度为30μmol/L时比其他浓度的细胞抑制率大。共培养组、c-Met阻滞剂组和RhoA阻滞剂组细胞中RhoA mRNA和蛋白表达水平均显著低于空白对照组，且以 RhoA 阻滞剂组最为显著。各组肝细胞生长因子浓度均随着时间的延长出现逐渐下降的情况，肝细胞生长因子激活物浓度均随着时间的延长出现逐渐上升的情况，且 c-Met 阻滞剂组变化最为显著。随着时间的延长，各组肝星状细胞凋亡率均出现逐渐上升的情况，其中RhoA阻滞剂组凋亡率最高，c-Met阻滞剂组最低。表明骨髓间充质干细胞会参与到肝星状细胞的凋亡机制中，并通过对肝细胞生长因子的激活以及Rho通路的下调达到促进肝星状细胞凋亡的目的。
揹景：控製肝星狀細胞的激活和增殖是抗肝纖維化研究的重點，人類或鼠的骨髓間充質榦細胞可通過徬分泌的肝細胞生長因子誘導肝星狀細胞凋亡。目的：探討骨髓間充質榦細胞在肝星狀細胞凋亡中的參與機製。方法：構建共培養體繫，于半透膜上下層在細胞培養闆上對肝星狀細胞和骨髓間充質榦細胞進行接種和培養，單純肝星狀細胞組設為空白對照組，共培養的肝星狀細胞和骨髓間充質榦細胞組為共培養組，以3 mg/L c-Met阻滯劑榦預的肝星狀細胞和骨髓間充質榦細胞為c-Met阻滯劑組，以3 mg/L RhoA阻滯劑榦預的肝星狀細胞和骨髓間充質榦細胞為RhoA阻滯劑組。結果與結論：c-Met阻滯劑質量濃度為3.0 mg/L，RhoA阻滯劑濃度為30μmol/L時比其他濃度的細胞抑製率大。共培養組、c-Met阻滯劑組和RhoA阻滯劑組細胞中RhoA mRNA和蛋白錶達水平均顯著低于空白對照組，且以 RhoA 阻滯劑組最為顯著。各組肝細胞生長因子濃度均隨著時間的延長齣現逐漸下降的情況，肝細胞生長因子激活物濃度均隨著時間的延長齣現逐漸上升的情況，且 c-Met 阻滯劑組變化最為顯著。隨著時間的延長，各組肝星狀細胞凋亡率均齣現逐漸上升的情況，其中RhoA阻滯劑組凋亡率最高，c-Met阻滯劑組最低。錶明骨髓間充質榦細胞會參與到肝星狀細胞的凋亡機製中，併通過對肝細胞生長因子的激活以及Rho通路的下調達到促進肝星狀細胞凋亡的目的。
배경：공제간성상세포적격활화증식시항간섬유화연구적중점，인류혹서적골수간충질간세포가통과방분비적간세포생장인자유도간성상세포조망。목적：탐토골수간충질간세포재간성상세포조망중적삼여궤제。방법：구건공배양체계，우반투막상하층재세포배양판상대간성상세포화골수간충질간세포진행접충화배양，단순간성상세포조설위공백대조조，공배양적간성상세포화골수간충질간세포조위공배양조，이3 mg/L c-Met조체제간예적간성상세포화골수간충질간세포위c-Met조체제조，이3 mg/L RhoA조체제간예적간성상세포화골수간충질간세포위RhoA조체제조。결과여결론：c-Met조체제질량농도위3.0 mg/L，RhoA조체제농도위30μmol/L시비기타농도적세포억제솔대。공배양조、c-Met조체제조화RhoA조체제조세포중RhoA mRNA화단백표체수평균현저저우공백대조조，차이 RhoA 조체제조최위현저。각조간세포생장인자농도균수착시간적연장출현축점하강적정황，간세포생장인자격활물농도균수착시간적연장출현축점상승적정황，차 c-Met 조체제조변화최위현저。수착시간적연장，각조간성상세포조망솔균출현축점상승적정황，기중RhoA조체제조조망솔최고，c-Met조체제조최저。표명골수간충질간세포회삼여도간성상세포적조망궤제중，병통과대간세포생장인자적격활이급Rho통로적하조체도촉진간성상세포조망적목적。
BACKGROUND:Control of hepatic stelate cel activation and proliferation is the focus of developing strategies against liver fibrosis. Human or murine bone marrow mesenchymal stem cels can induce apoptosis of hepatic stelate cels through paracrine of hepatocyte growth factors. OBJECTIVE:To explore the mechanism by which bone marrow mesenchymal stem cels participate in apoptosis of rat hepatic stelate cels. METHODS:Hepatic stelate cels and bone marrow mesenchymal stem cels were seeded and co-cultured in the upper and lower chambers in a co-culture system, serving as a co-culture group. In the blank control group, only hepatic stelate cels were involved. In the c-Met inhibitor group, hepatic stelate cels and bone marrow mesenchymal stem cels were treated with 3 mg/L C-Met inhibitor. In the RhoA inhibitor group, both kinds of cels were treated with 3 mg/L RhoA inhibitor. RESULTS AND CONCLUSION:The concentration of c-Met inhibitor was 3.0 mg/L. RhoA inhibitor at 30μmol/L exhibited a greater inhibitory effect than at other concentrations. RhoA mRNA and protein expression in the co-culture, c-Met inhibitor and in particular RhoA inhibitor groups was obviously greater than in the blank control group. Hepatocyte growth factor concentration in each group was gradualy decreased with time, hepatocyte growth factor activator concentration in each group was gradualy increased with time, and the changes were most obvious in the c-Met inhibitor group. Apoptosis rate of hepatic stelate cels in each group was gradualy increased with time, and highest apoptosis rate appeared in the RhoA inhibitor group, and lowest apoptosis rate in the c-Met inhibitor group. These findings suggest that bone marrow mesenchymal stem cels participate in and promote the apoptosis of hepatic stelate cels by activating hepatocyte growth factors and downregulating Rho activity.
