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目的:研究肝素与大鼠肝星状细胞(hepatic stellate cells,HSC)作用后转化生长因子β1 (transforming growth factorβ1,TGF-β1)和Ⅰ型胶原表达的变化及意义.方法:大鼠肝星状细胞以1×10~8/L浓度接种于96孔培养板,每孔100μL.实验分组为肝素Ⅰ组、肝素Ⅱ组、肝素Ⅲ组,加入肝素使各组培养液中肝素浓度分别是10,100,1000 mg/L,加生理盐水为对照组(每组6孔重复3次)培养48 h.培养终止后吸取上清液-20℃冰冻保存,ELISA法检测其上清液TGF-β1和Ⅰ型胶原水平,MTT法观察细胞增殖情况.结果:肝素Ⅱ组和Ⅲ组HSC培养上清液TGF-β1水平均显著低于对照组(4.59±1.27 ng/L,3.34±1.13 ng/L vs 5.95±1.72 ng/L,P均<0.01),肝素各组Ⅰ型胶原水平均低于对照组(87.20±9.30 ng/L,73.17±12.04 ng/L vs 95.61±12.55 ng/L,63.31±10.93 ng/L vs 95.61±12.55 ng/L,P均<0.05),肝素Ⅲ组平均吸光度低于肝素Ⅰ组和对照组(0.29±0.07 vs 0.42±0.12,0.46±0.17,P均<0.05).结论:大鼠肝星状细胞在肝素作用下TGF-β1和Ⅰ型胶原分泌受抑制,其增殖减少.
OBJECTIVE: To study the changes of transforming growth factor β1 (TGF-β1) and type Ⅰ collagen after heparin and rat hepatic stellate cells (HSC) The cells were inoculated into 96-well plates at the concentration of 1 × 10-8 / L with 100μL per well.The experimental groups were heparin Ⅰ, heparin Ⅱ and heparin Ⅲ, heparin concentration was 10,100, 1000 mg / L, normal saline was added to the control group (6 wells repeated 3 times each) for 48 h, then the supernatant was frozen and stored at -20 ℃ after the termination of culture, and the supernatant TGF-β1 and type Ⅰ Collagen level and MTT assay were used to observe the cell proliferation.Results: The levels of TGF-β1 in HSC culture supernatant of group Ⅱ and group Ⅲ were significantly lower than those of control group (4.59 ± 1.27 ng / L, 3.34 ± 1.13 ng / L vs 5.95 ± 1.72 ng / L, P <0.01). The levels of type I collagen in heparin group were significantly lower than those in control group (87.20 ± 9.30 ng / L, 73.17 ± 12.04 ng / L vs 95.61 ± 12.55 ng / L, 63.31 ± 10.93 ng / L vs 95.61 ± 12.55 ng / L, P <0.05). The average absorbance of heparin group was lower than that of heparin group Ⅰ and control group (0.29 ± 0.07 vs 0.42 ± 0.12,0.46 ± 0.17, P <0.05) .Conclusion : Rat hepatic stellate cells inhibit the secretion of TGF-β1 and type Ⅰ collagen under the action of heparin, and their proliferation is reduced.