论文部分内容阅读
目的:研究蛇葡萄素对大鼠肝星状细胞(HSC-T6)的增殖及胶原蛋白、细胞因子生成的影响。方法:对HSC-T6增殖的影响:用含10%胎牛血清的高糖DMEM培养液制备成HSC-T6细胞3×104个/mL细胞悬液100μL/孔接种于96孔培养板。设细胞对照组、秋水仙碱1.0 mg·L-1组和100.0,75.0,50.0,25.0,12.5 mg·L-15个不同质量浓度蛇葡萄素组,置37℃5%CO2培养48 h。采用MTT法检测蛇葡萄素对大鼠肝细胞HSC-T6增殖的影响。对HSC-T6生成Ⅰ,Ⅲ,Ⅳ型胶原蛋白及细胞因子(TGF-β1,PDGF)含量的影响:取终质量浓度分别为50.0,25.0,12.5 mg·L-1蛇葡萄素,3×104个/mL细胞悬液100μL/孔细胞液接种于96孔培养板。阳性组秋水仙碱的质量浓度为1.0 mg·L-1。置37℃5%CO2培养48 h。ELISA法检测HSCT6细胞生成的胶原蛋白(Ⅰ,Ⅲ,Ⅳ)、转化生长因子(TGF-β1)及血小板衍生因子(PDGF)含量。结果:蛇葡萄素100.0,75.0mg·L-1对HSC-T6细胞的增殖有明显抑制作用,其无毒浓度为50.0 mg·L-1。50 mg·L-1的蛇葡萄素可使HSC-T6细胞生成的Ⅰ,Ⅲ,Ⅳ型胶原蛋白,TGF-β1,PDGF含量明显低于细胞对照组(P<0.05),25 mg·L-1的蛇葡萄素也可使HSC-T6细胞生成的Ⅲ型胶原蛋白含量明显低于细胞对照组(P<0.05)。结论:蛇葡萄素能抑制HSC-T6增殖并可使HSC-T6生成的Ⅰ,Ⅲ,Ⅳ型胶原蛋白,TGF-β1,PDGF含量降低。
Objective: To study the effect of snake grape wine on the proliferation, collagen and cytokine production of rat hepatic stellate cells (HSC-T6). Methods: Effects on proliferation of HSC-T6: Prepare HSC-T6 cell suspension containing 100 cells / mL of HSC-T6 cells in 96-well culture plate by high glucose DMEM medium containing 10% fetal bovine serum. The cell control group, colchicine 1.0 mg · L-1 group and 100.0,75.0,50.0,25.0,12.5 mg · L-15 different concentrations of the snake grape juice group were cultured in 37 ℃ 5% CO2 for 48 h. MTT assay was used to detect the effect of strychnine on the proliferation of rat hepatocyte HSC-T6. The effect of HSC-T6 on the production of collagen type Ⅰ, Ⅲ, Ⅳ and TGF-β1, PDGF in HSC-T6: The final concentrations were 50.0,25.0,12.5 mg · L-1 and 3 × 104 / ML cell suspension 100 μL / well of cell suspension in 96-well culture plate. The concentration of colchicine in the positive group was 1.0 mg · L -1. Placed in 37 ℃ 5% CO2 48 h. The contents of collagen (Ⅰ, Ⅲ, Ⅳ), transforming growth factor (TGF-β1) and platelet-derived factor (PDGF) in HSCT6 cells were detected by ELISA. RESULTS: 100μmol·L-1, 0.050mg · L-1 of S. pombe obviously inhibited the proliferation of HSC-T6 cells. The non-toxic concentration of 50μmol·L-1.50 mg · L- The levels of TGF-β1 and PDGF in type Ⅰ, type Ⅲ and type Ⅳ collagen-T6 cells were significantly lower than those in the cell control group (P <0.05). The 25 mg · L-1 sphingosine also induced HSC-T6 cells Of type Ⅲ collagen content was significantly lower than the cell control group (P <0.05). CONCLUSION: The combination of amylopsin and HSC-T6 can inhibit the proliferation of HSC-T6 and reduce the content of collagen type I, III and IV, TGF-β1 and PDGF.