肝纤维化大鼠药物血清对肝星状细胞增殖及活性型TGFβ_1含量的影响

Eeffects of Drug Sera from Liver Fibrosis Rats on Proliferation of Hepatic Stellate Cells and Active Type TGFβ_1 Content

目的:观察肝纤维化大鼠药物血清对肝星状细胞(hepatic stellate cells,HSC)增殖及活性型TGFβ_1,含量的影响。方法:采用二甲基亚硝胺(dimethylnitrosamine,DMN)造成大鼠肝纤维化模型,经丹参酚酸B盐(Salvianolic-acid B,SA-B)与虫草多糖(Cordyceps sinensis polysaccharide,CP)联合治疗后采集大鼠血清,通过体外培养HSC,运用比色法、噻唑蓝(MTT)法及生物学检测法来观察药物血清对HSC增殖及活性型TGFβ_1含量的影响。结果:①细胞培养上清乳酸脱氢酶(LDH)活力测定显示,模型大鼠血清和药物血清无细胞毒性,正常大鼠血清组、模型大鼠血清组、药物血清组细胞培养上清LDH活力分别为(95±7.88)U/L,(72.2±11.42)U/L,(48.13±62.16)U/L,正常大鼠血清组与模型大鼠血清组比,差异有统计学意义(P<0.05)。②在对HSC增殖的影响方面,正常大鼠血清与模型大鼠血清之间的差异无统计学意义。与模型大鼠血清比较,药物血清能抑制HSC增殖(P<0.05)。③与模型大鼠血清比较,正常大鼠血清与药物血清均能减少活性型TGFβ_1含量,差异均有统计学意义(P<0.05)。结论:DMN肝纤维化大鼠血清与正常大鼠血清对HSC的影响不尽相同;SA-B合CP能抑制HSC增殖,并能减少活性型TGFβ_1含量。

Objective: To observe the effects of drug sera from liver fibrosis rats on proliferation of hepatic stellate cells (HSC) and active type TGFβ1 content. Methods: The fibrotic model was induced by Dimethylnitrosamine (DMN) in rats, treated with the association of Salvianolic-acid B (SA-B) and Cordyceps sinensis polysaccharide (CP). Then sera collected from rats were added into activated HSC culture medium. In order to observe the effects of drug sera on proliferation of HSC and active type TGFβ1 content, lactic dehydrogenase (LDH) activity in culture medium was measured with colorimeter assay, also MTT method and biological method were adopted. Results: (1)LDH activity in culture medium showed that sera from model rats and drug sera had no toxicity to HSC. LDH activity of normal rat sera group was (95±7.88) U/L, LDH activity of model rat sera group was (72.2± 11.42) U/L, LDH activity of drug sera group was (48.13±62.16) U/L. Normal rat sera group was compared with model rat sera group, the difference between them had statistic significance, P<0.05.(2)The difference of the effect on proliferation of HSC between normal rat sera and model rat sera had no obvious significance. Compared with model rat sera, drug sera could inhibit proliferation of HSC, P<0.05. (3)Compared with model rat sera, either normal rat sera or drug sera could decrease active type TGFβ1 content, P<0.05. Conclusion: It is different between DMN liver fibrosis rat sera and normal rat sera that the effects on HSC; the association of SA-B and CP could inhibit proliferation of HSC and decrease active type TGFβ1 content.