【摘要】 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。更多还原

【Abstract】 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,25 mmol / 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 48 h. Inverted microscopy was used to observe the morphological changes in high glucose- induced HK- 2 cells. Expression of ɑ- smooth muscle actin( ɑ- 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 ɑ- 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.更多还原