基于肾小管上皮细胞的HIF-1α/STAT3/IL-6通路激活成纤维细胞探讨银杏叶提取物抑制急性顺铂肾损伤慢性化的作用机制

急性顺铂肾损伤慢性化机制未阐明且缺乏有效防治措施。我们发现，银杏叶提取物（EGb）抑制大鼠顺铂肾间质纤维化和肾HIF-1α；顺铂使肾小管上皮细胞（RTEC）HK-2产生HIF-1α、STAT3和分泌IL-6，该HK-2使共培养成纤维细胞α-SMA升高；抑制HK-2的HIF-1α可降低STAT3、IL-6和共培养成纤维细胞α-SMA；外来IL-6活化成纤维细胞；推测RTEC通过HIF-1α/STAT3/IL-6活化成纤维细胞为肌成纤维细胞，EGb抑制HIF-1α而抑制该通路进而抑制肾间质纤维化。本项目拟建立沉默HIF-1α、IL-6基因的HK-2与成纤维细胞共培养模型，以及大鼠模型，用顺铂、EGb干预，检测细胞活化和纤维化、通路指标α-SMA、ColⅠ、HIF-1α、STAT3、IL-6等，从RTEC的HIF-1α/STAT3/IL-6激活成纤维细胞探讨EGb抑制急性顺铂肾损伤慢性化的机制。

Acute kidney injury induced by cisplatin can lead to chronicity and its mechanism is still not elucidated, and there is a shortage of effective preventive and therapeutic measures for it. We found that ginkgo biloba extract (EGb) could inhibit the cisplatin-induced renal interstitial fibrosis, and reduced the HIF-1α level in renal tissue. Renal tubular epithelial cell (RTEC) line HK-2 produced HIF-1α and STAT3, and secreted IL-6 induced by cisplatin. Moreover, the damaged HK-2 elevated the α-SMA level in the co-cultured fibroblasts. Additionally, inhibition the HIF-1α of HK-2 could reduce its STAT3 level and the secretion of IL-6, as well as the α-SMA level of the co-cultured fibroblast. Foreign IL-6 could activate fibroblasts. Therefore, it was speculated that RTEC activated fibroblasts into myofibroblasts by HIF-1α/STAT3/IL-6 pathway, and EGb inhibited this pathway by inhibition HIF-1α and in turn to inhibit renal interstitial fibrosis induced by cisplatin. This project intends to establish a co-culture model using HK-2 cells (silence HIF-1α gene or IL-6 gene) and fibroblasts, and establish a rat model, and then cisplatin and EGb were used for intervention. Cell activation, fibrosis and pathway indicators such as α-SMA, Col I, HIF-1α, STAT3, IL-6 will been detected, to investigate the mechanism of EGb in inhibiting chronicty of cisplatin-induced acute renal injury base on RTEC activated fibroblasts through HIF-1α/STAT3/IL-6.