首页|根皮素通过抑制FAK改善肾小管间质纤维化

根皮素通过抑制FAK改善肾小管间质纤维化

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目的 探究根皮素能否通过抑制黏着斑激酶(FAK)改善肾小管间质纤维化。方法 肾小管上皮细胞NRK-52E经TGF-β1(10 ng/mL)诱导建立肾纤维化细胞模型，并构建单侧输尿管梗阻(UUO)肾间质纤维化小鼠模型。通过免疫荧光及Western blot法检测细胞及小鼠肾脏组织中α-SMA、FN的蛋白表达水平，明确根皮素体内外抗肾纤维化作用;检测实验小鼠血清中肌酐(Cr)和尿素氮(BUN)水平，分析根皮素的肾保护作用;以苏木精-伊红(HE)染色和Masson染色观察根皮素对UUO小鼠肾脏病理学改变和胶原纤维沉积的影响;Western blot检测TGF-β1诱导的NRK-52E细胞和UUO小鼠肾组织中FAK、p-FAK的蛋白表达水平;通过分子对接预测根皮素与FAK蛋白的潜在结合模式和结合能力;使用FAK抑制剂PND-1186，探究根皮素改善肾间质纤维化的作用机制。结果 根皮素可逆转TGF-β1所致的肾小管上皮细胞及UUO小鼠肾组织中α-SMA、FN的蛋白表达水平升高，且显著降低UUO小鼠血清中Cr、BUN水平，改善UUO小鼠肾小管扩张、炎性细胞浸润及间质胶原沉积;根皮素可以显著抑制UUO小鼠肾组织中FAK、p-FAK蛋白的表达水平，分子对接提示根皮素与FAK蛋白之间具有良好相互作用。根皮素与PND-1186均可改善TGF-β1诱导的NRK-52E细胞纤维化。结论 根皮素通过抑制FAK信号通路在体内外发挥抗肾间质纤维化作用。
Phloretin Improves Tubulointerstitial Fibrosis by Inhibiting FAK
Objective To explore whether phloretin alleviates tubulointerstitial fibrosis by inhibiting the focal adhesion ki-nase(FAK)signaling pathway.Methods Renal tubular epithelial cell NRK-52E was induced by TGF-β1(10 ng/mL)to establish fibrosis model and unilateral ureteral obstruction(UUO)mouse model.The protein expression levels of α-SMA,FN in cells and mice kidneys were tested by Western blotting or immunofluorescence to clarify the anti-fibrosis effect of phloretin in vivo and in vitro.The serum creatinine(Cr)and blood urea nitrogen(BUN)levels were measured to indicate the renal protective effect of phloretin on UUO mice.The pathological changes and collagen deposition in kidney of mice were observed by HE and Masson staining.Protein expression levels of FAK and p-FAK in cells and mouse kidney tissues were detected by Western blotting.The potential binding pattern and binding ability of phloretin to FAK protein were predicted via molecular docking,and the FAK in-hibitor PND-1186 was employed to investigate the specific mechanism of phloretin in alleviating renal interstitial fibrosis.Results Phloretin reversed the increasing trend of expression levels of α-SMA and FN induced by TGF-β1 in renal tubular epithelial cells and UUO mice,and the serum levels of Cr and BUN were significantly reduced in UUO mice.Renal tubular dilation,inflamma-tory cell infiltration and interstitial collagen deposition were improved in UUO mice.The expression levels of FAK and p-FAK proteins were significantly reduced in UUO mice kidney tissue after phloretin treatment.It was suggested by molecular docking that there was a correlation between phloretin and FAK proteins.Both phloretin and PND-1186 improved TGF-β1-induced NRK-52E cell fibrosis.Conclusion Phloretin alleviates renal fibrosis by suppressing FAK in vivo and in vitro.

phloretinunilateral ureteral obstructionrenal protectiontubulointerstitial fibrosisfocal adhesion kinase

程新瑶、曹文洁、钱永帅、刘丽、余惠凡、王晓菲、方梦淇、李飞

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湖北医药学院药学院

武当特色中药研究湖北省重点实验室

湖北医药学院生物医药研究院,十堰 442000

根皮素 单侧输尿管梗阻 肾保护 肾间质纤维化 黏着斑激酶

湖北省卫生健康委青年人才项目湖北省教育厅重点项目湖北医药学院研究生科技创新项目湖北医药学院湖北省高等学校优势特色学科群资助项目(十四五)湖北医药学院大学生创新创业训练计划湖北医药学院大学生创新创业训练计划

WJ2021Q010D20192101YC20230672022BMXKQT1X202310929009X202310929010

2024

10.3870/j.issn.1672-0741.23.10.007

华中科技大学学报(医学版)
华中科技大学

华中科技大学学报(医学版)

CSTPCD北大核心
影响因子：1.443
ISSN：1672-0741
年,卷(期)：2024.53(2)
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