大蒜素对高糖诱导人肾小管上皮细胞转分化的影响

Effects of Allicin on Human Tubular Epithelial-myofibroblast Transdifferentiation Induced by High Glucose

目的探讨大蒜素对高糖诱导的人肾小管上皮细胞(HK-2)转分化的影响。方法复苏并传代培养HK-2细胞,分为正常糖对照组(NG,5.5mmol/L)、高糖组(HG,25mmol/L)、高糖+不同剂量的大蒜素干预组(HG+2.5、5、10、20μg/ml Allicn)、高糖+JAK2抑制剂AG490组(HG+10μmol/L AG490)。倒置显微镜下观察高糖对细胞形态的影响,细胞免疫荧光检测α平滑肌肌动蛋白(α-SMA)、E钙黏蛋白(E-cadherin)、Ⅰ型胶原蛋白(collagenⅠ)的表达变化,荧光定量PCR检测TGF-β1mRNA的表达变化,Western blot法检测TGF-β1、p-STAT3、STAT3蛋白的表达变化。结果与正常对照组相比,高糖刺激后的HK-2细胞的形态发生明显的长梭形改变,上皮细胞标志物E-cadherin表达明显减少,而间充质细胞标志物ɑ-SMA及collagenⅠ的表达明显升高(P<0.01);TGF-β1及p-STAT3的表达明显增高(P<0.05);而加入大蒜素或JAK2抑制剂AG490后,均能不同程度抑制高糖诱导的HK-2细胞转分化,而且大蒜素可明显抑制TGF-β1及p-STAT3的表达,以20μg/ml尤为显著(P<0.05)。结论大蒜素可能通过抑制JAK2/STAT3信号通路活化,从而抑制高糖诱导的HK-2细胞转分化。

Objective To investigate the effect of Allicin on high glucose-induced human renal proximal tubular epithelial myofibroblast transdifferentiation（TEMT）.Methods Cultured HK-2 cells were divided into 7 groups：normal glucose （NG,5.5mmol/L）,high glucose （HG,25mmol/L）,high glucose with Allicin treatment （HG ＋ 2.5,5,10,20μg/ml Allicin） and high glucose with the JAK2 inhibitor AG490 group （HG ＋ 10μmol/L AG490）.The cells were cultured for 48h.Inverted microscopy was used to observe the morphological changes in high glucose-induced HK-2 cells.Expression of α-smooth muscle actin （α-SMA）,E-cadherin,collagen Ⅰwere detected by immunofluorescence assay,the expression of TGF-β1 mRNA were measured by real-time PCR,and that of TGF-β1 、p-STAT3、STAT3 were measured by Western blot.Results After incubated with high glucose,HK-2 cells turned into long spindle-shape compared with NG group.The expression of epithelial marker E-cadherin was significantly decreased,while those of Collagen Ⅰ and the mesenchymal markers α-SMA were significantly increased （P ＜ 0.01）.The expression of TGF-β1 and p-STAT3 were significantly increased （P ＜ 0.05）.The administration of Allicin or AG490 obviously inhibited EMT （especially at 20μg/ml,P ＜ 0.01） in HK-2 cells induced by high glucose,while Allicin markedly decreased the expression of TGF-β1 and p-STAT3 （especially at 20μg/ml,P ＜ 0.05）.Conclusion Allicin may inhibit tubular epithelial-myofibroblast transdifferentiation via at least regulating JAK2/ p-STAT3 signaling pathway.