虫草菌液拮抗人白蛋白致HK-2细胞增殖及TGF-β1、FN表达的影响

Hirsutella sinensis antagonizes the effects of human albumin on the expression of TGF-β1,FN and the proliferation in HK-2 cells

目的初步探讨虫草菌液能否拮抗人白蛋白致HK-2细胞的促纤维化效应。方法人白蛋白组(albumin,ALB 5g/L)加或不加虫草菌液(hirsu-tella sinensis,HS 10mg/L)与HK-2细胞孵育,然后用四甲基偶氮唑盐(MTT)比色法测定细胞增殖水平;使用酶联免疫吸附法(ELISA)检测细胞上清FN、TGF-β1的分泌水平。采用逆转录多聚酶链式反应(RT-PCR)半定量分析HK-2中TGF-β1及Fn的mRNA表达。结果虫草菌液可抑制人白蛋白所致HK-2增殖(P<0.01),虫草菌液干预人白蛋白孵育的HK-2,可使TGF-β1和Fn mRNA和分泌及表达明显下调(P<0.05)。结论虫草菌液可抑制人白蛋白刺激的HK-2细胞增殖和TGF-β1、FN表达。

[Objective] To investigate the effect of HirsuteUa sinensis （HS） on tubulointerstitial fibrosis and its possible mechanism. [Methods] HK-2 ceils were incubated with medium alone, medium with HS 10 mg/L, medium with albumin 5 g/L or medium with albumin 5 g/L and HS 10 mg/L, respectively. The capacity of proliferation of HK-2 was assessed by tetrazolium salt colofimetry assay （MTT assay）. ELISA was used to measure the secretion of FN, TGF-β1 at 24 hours. The mRNA expressions of TGF-β1 and Fn were determined by semi-quantification reverse tran-scfiptive PCR （RT-PCR）. [Results] The proliferative capacity in HS groups was significantly lower than group ALB 5.0 g/L at 24 hours （P 〈0.01）. Obvious decrease of TGF-β1 was found in HS groups compared with group stimulated with albumin 5 g/L （P 〈0.05）. Exposure of HK-2 to HS reduced the Fn secretion （P 〈0.05）. [Conclusion] HS can inhibit effect on cellular proliferation. HS reduced the expressin of TGF-β1 in HK-2 stimulated by albumin 5 g/L, and reduced Fn production. These may be the new mechanisms of HS to postpone the tubulointerstitial fibrosis.