银杏叶提取物通过下调肾小管上皮细胞中 HIF-1α/STAT3/IL-6 通路抑制肾成纤维细胞的激活，减轻顺铂诱导的肾间质纤维化。
Ginkgo biloba extract attenuates cisplatin-induced renal interstitial fibrosis by inhibiting the activation of renal fibroblasts through down-regulating the HIF-1α/STAT3/IL-6 pathway in renal tubular epithelial cells.
机构信息
Department of Pharmacy, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China.
Department of Pharmacy, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China.
出版信息
Phytomedicine. 2023 Jul;115:154809. doi: 10.1016/j.phymed.2023.154809. Epub 2023 Apr 6.
BACKGROUND
Activation of renal fibroblasts into myofibroblasts plays an important role in promoting renal interstitial fibrosis (RIF). Ginkgo biloba extract (EGb) can alleviate RIF induced by cisplatin (CDDP).
PURPOSE
To elucidate the effect of EGb treatment on cisplatin-induced RIF and reveal its potential mechanism.
METHODS
The two main active components in EGb were determined by high-performance liquid chromatography (HPLC) analysis. Rats were induced by CDDP and then treated with EGb, 2ME2 (HIF-1α inhibitor) or amifostine. After HK-2 cells and HIF-1α siRNA HK-2 cells were treated with CDDP, EGb or amifostine, the conditioned medium from each group was cultured with NRK-49F cells. The renal function of rats was detected. The renal damage and fibrosis were evaluated by H&E and Masson trichrome staining. The IL-6 content in the cell medium was detected by ELISA. The expression levels of indicators related to renal fibrosis and signaling pathway were examined by western blotting and qRT-PCR.
RESULTS
HPLC analysis showed that the contents of quercetin and kaempferol in EGb were 36.0 μg/ml and 45.7 μg/ml, respectively. In vivo, EGb and 2ME2 alleviated renal damage and fibrosis, as well as significantly decreased the levels of α-SMA, HIF-1α, STAT3 and IL-6 in rat tissues induced by CDDP. In vitro, the levels of HIF-1α, STAT3 and IL-6 were significantly increased in HK-2 cells and HIF-1α siRNA HK-2 cells induced by CDDP. Notably, HIF-1α siRNA significantly decreased the levels of HIF-1α, STAT3 and IL-6 in HK-2 cells, as well as the IL-6 level in medium from HK-2 cells. Additionally, the α-SMA level in NRK-49F cells was significantly increased after being cultured with conditioned medium from HK-2 cells or HIF-1α siRNA HK-2 cells exposed to CDDP. Furthermore, exogenous IL-6 increased the α-SMA level in NRK-49F cells. Importantly, the expression levels of the above-mentioned indicators were significantly decreased after the HK-2 cells and HIF-1α siRNA HK-2 cells were treated with EGb.
CONCLUSION
This study revealed that EGb improves CDDP-induced RIF, and the mechanism may be related to its inhibition of the renal fibroblast activation by down-regulating the HIF-1α/STAT3/IL-6 pathway in renal tubular epithelial cells.
背景
肾成纤维细胞向肌成纤维细胞的激活在促进肾间质纤维化（RIF）中起着重要作用。银杏叶提取物（EGb）可减轻顺铂（CDDP）引起的 RIF。
目的
阐明 EGb 对顺铂诱导的 RIF 的作用，并揭示其潜在机制。
方法
采用高效液相色谱法（HPLC）分析 EGb 中的两种主要活性成分。用 CDDP 诱导大鼠，然后用 EGb、2ME2（HIF-1α抑制剂）或氨磷汀治疗。用 CDDP、EGb 或氨磷汀处理 HK-2 细胞和 HIF-1α siRNA HK-2 细胞后，将各组的条件培养基培养 NRK-49F 细胞。检测大鼠肾功能。通过 H&E 和 Masson 三色染色评估肾损伤和纤维化。通过 ELISA 检测细胞培养基中 IL-6 的含量。通过 Western blot 和 qRT-PCR 检测与肾纤维化和信号通路相关的指标的表达水平。
结果
HPLC 分析显示，EGb 中槲皮素和山奈酚的含量分别为 36.0μg/ml 和 45.7μg/ml。体内，EGb 和 2ME2 减轻了由 CDDP 诱导的大鼠肾脏损伤和纤维化，并且显著降低了大鼠组织中由 CDDP 诱导的α-SMA、HIF-1α、STAT3 和 IL-6 的水平。体外，用 CDDP 诱导的 HK-2 细胞和 HIF-1α siRNA HK-2 细胞中 HIF-1α、STAT3 和 IL-6 的水平显著升高。值得注意的是，HIF-1α siRNA 显著降低了 HK-2 细胞中 HIF-1α、STAT3 和 IL-6 的水平，以及 HK-2 细胞培养基中的 IL-6 水平。此外，用 CDDP 处理 HK-2 细胞或 HIF-1α siRNA HK-2 细胞的条件培养基培养 NRK-49F 细胞后，NRK-49F 细胞中的α-SMA 水平显著增加。此外，外源性 IL-6 增加了 NRK-49F 细胞中的α-SMA 水平。重要的是，用 EGb 处理 HK-2 细胞和 HIF-1α siRNA HK-2 细胞后，上述指标的表达水平显著降低。
结论
本研究表明 EGb 改善了由 CDDP 引起的 RIF，其机制可能与其通过下调肾小管上皮细胞中的 HIF-1α/STAT3/IL-6 通路抑制肾成纤维细胞的激活有关。
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