虫草菌液拮抗马兜铃酸对人近端肾小管上皮细胞的作用

目的 探讨虫草菌液能否拮抗马兜铃酸诱发的人近端肾小管上皮细胞系(HKC)的促纤维化效应。 方法 马兜铃酸钠盐(AA-Na，40 mg/L)加或不加虫草菌液(10 mg/L)与HKC孵育(孵育12 h检测mRNA表达，孵育36 h检测蛋白质表达)，然后检测HKC中转化生长因子-β1(TGF-β1)、结缔组织生长因子(CTGF)、金属蛋白酶1组织抑制物(TIMP-1)以及纤溶酶原激活物抑制物1(PAI-1)的mRNA(RT-PCR法)和相应蛋白质(CTGF表达采用免疫印迹法，其余采用ELISA方法)表达。结果 AA-Na能显著上调HKC对TGF-β1、CTGF、TIMP-1、PAI-1的表达，与对照组比较，mRNA表达分别上调1.24、1.31、1.27及1.36倍，蛋白质表达分别上调2.50、1.75、2.13及1.46倍，P均<0.05。加虫草菌液后，上述TGF-β1、CTGF及TIMP-1的高表达被显著抑制，与AA-Na组比较，mRNA表达的抑制率分别为12%、20%及17%；蛋白质表达的抑制率分别为25%、20%及37%，P均<0.05，但未能抑制PAI-1的高表达(P > 0.05)。结论 虫草菌液可下调AA-Na刺激的HKC促细胞外基质(ECM)合成因子(TGF-β1、CTGF)及抗ECM降解因子(TIMP-1)的表达。

Hirsutella sinensis antagonizes the effect of aristolochic acid on human proximal tubular epithelial cells

Objective To study whether Hirsutella sinensis (HS) can antagonize aristolochic acid (AA) induced fibrogenesis on human proximal tubular epithelial cells (HKC). Methods HKC were incubated with medium alone， medium with HS 10 mg/L， medium with AA-Na 40 mg/L or medium with AA-Na 40 mg/L and HS 10 mg/L， respectively. After 12 h (for mRNA) or 36 h (for protein)，cells were lysed，and the mRNA and protein expression level of transforming growth factor-β1 (TGF-β1)， connective tissue growth factor (CTGF)， tissue inhibitor of metalloproteinase-1 (TIMP-1)and plasminogen activator inhibitor-1 (PAI-1) of HKC were measured by RT-PCR， ELISA (for TGF-β1， TIMP-1 and PAI-1) and Western blotting (for CTGF)， respectively. Results The mRNA and protein expression of TGF-β1，PAI-1，CTGF and TIMP-1 were significantly up-regulated by AA-Na 40 mg/L. Compared with the control group， the mRNA expression of TGF-β1，CTGF，TIMP-1 and PAI-1 was up-regulated to 1.24，1.31，1.27 and 1.36 times，respectively(P﹤0.05). Their protein expression was up-regulated to 2.50，1.75，2.13 and 1.46 times， respectively(P﹤0.05). HS significantly inhibited the up-regulation of TGF-β1，CTGF，and TIMP-1 expression induced by AA. Compared with the AA-Na stimulation group， the inhibition rates of TGF-β1，CTGF and TIMP-1 mRNA expression in the HS group were 12%，20% and 17%， respectively(P﹤0.05)， and the inhibition rates of their protein expression were 25%，20%， and 37%， respectively (P﹤0.05).HS did not inhibit the up-regulation of PAI-1 expression induced by AA. Conclusion The AA- induced fibrogenic effects on HKC can be antagonized by HS， which may be through down-regulating the factors promoting extra cellular matrix (ECM) synthesis (TGF-β1 and CTGF) and inhibiting ECM degradation (TIMP-1).