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目的:研究银杏叶提取物(extract of ginkgo biloba,EGB)对转化生长因子β1(transforming growth factor-β1 TGF-β1)诱导的人肾小管上皮细胞HK2(human tubular epithelial cells)转分化的影响及其机制。方法:复苏并传代培养HK2细胞,应用10μg/LTGF-β1诱导HK2细胞向间充质的转分化,给予EGB干预。应用West-ern印记法检测HK2细胞E-钙黏蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、NADPH氧化酶p67phox以及超氧化物歧化酶(superoxide dismutase,SOD)的表达,检测细胞培养液上清中丙二醛(malondialdehyde,MDA)的含量。结果:EGB显著抑制TGF-β1诱导的E-cadherin下调(P<0.05)和α-SMA上调(P<0.05);EGB显著抑制TGF-β1诱导的p67phox表达增加(P<0.05),显著抑制TGF-β1诱导的SOD表达降低(P<0.05);同时,TGF-β1显著促进了MDA的产生(P<0.05),EGB可以部分抑制TGF-β1刺激的MDA浓度的升高(P<0.05)。结论:EGB显著抑制TGF-β1诱导的HK2细胞转分化,其机制可能是EGB抑制了TGF-β1诱导的p67phox上调并上调SOD的表达。
Objective: To study the effect of extract of ginkgo biloba (EGB) on transdifferentiation of human tubular epithelial cells (HK2) induced by transforming growth factor-β1 TGF-β1. mechanism. METHODS: HK2 cells were resuscitated and subcultured, and 10 μg/L TGF-β1 was used to induce transdifferentiation of HK2 cells into mesenchymal cells and EGB intervention was performed. Immunohistochemical staining of E-cadherin, α-smooth muscle actin (α-SMA), NADPH oxidase p67phox, and superoxide dismutase (superoxide dismutase) in HK2 cells was detected by Western blot. The expression of dismutase, SOD) was detected by measuring the content of malondialdehyde (MDA) in the supernatant of cell culture medium. RESULTS: EGB significantly inhibited TGF-β1-induced E-cadherin downregulation (P<0.05) and α-SMA upregulation (P<0.05); EGB significantly inhibited the expression of p67phox induced by TGF-β1 (P<0.05), and significantly inhibited TGF The expression of SOD induced by -β1 was decreased (P<0.05). At the same time, TGF-β1 significantly promoted the production of MDA (P<0.05). EGB partially inhibited the increase of MDA stimulated by TGF-β1 (P<0.05). CONCLUSION: EGB significantly inhibits the transdifferentiation of HK2 cells induced by TGF-β1. The mechanism may be that EGB inhibits the up-regulation of p67phox induced by TGF-β1 and up-regulates the expression of SOD.