金丝桃苷通过抑制YAP改善肾小管间质纤维化  

Hypericin Ameliorates Renal Tubulointerstitial Fibrosis via Inhibiting YAP

作　　者：王俊青 余惠凡 黄林生[2] 汪鲁洁 刘静雨 袁璐 李飞 Wang Junqing;Yu Huifan;Huang Linsheng(School of Pharmaceutical Sciences,Hubei University of Medicine,Shiyan 442000,China;Department of Hepatopancreatobiliary Surgery,Taihe Hos pital,Hubei Unriversity of Medicine,Shiyan 442000,China)

机构地区：[1]湖北医药学院药学院,十堰442000 [2]湖北医药学院附属太和医院肝胆胰外科,十堰442000

出　　处：《华中科技大学学报(医学版)》2025年第2期190-197,共8页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：湖北医药学院研究生科技创新项目(No.YC2024070,No.YC2024071,No.YC2024072);湖北医药学院“十四五”湖北省高等学校优势特色学科群(生物与医药)资助项目(No.2024BMXKQT1)。

摘　　要：目的明确金丝桃苷的抗肾纤维化作用并探究金丝桃苷是否能够通过抑制Yes相关蛋白(YAP)发挥缓解肾小管间质纤维化作用。方法将36只雄性小鼠随机分为假手术(Sham)组、假手术+120 mg/kg金丝桃苷组、单侧输尿管梗阻(UUO)模型组、金丝桃苷(30、60、120 mg/kg)治疗组,每组6只,除Sham组外,其余组小鼠经UUO建立肾间质纤维化模型,给药组小鼠给予相应剂量的金丝桃苷混悬液,Sham组和UUO模型组小鼠灌胃给予相应体积生理盐水,连续14 d。使用TGF-β1(10 ng/mL)诱导NRK-52E细胞构建细胞纤维化模型。采用Western blot法检测UUO小鼠肾组织及TGF-β1诱导的NRK-52E细胞中α-SMA、Fibronectin、CollagenⅠ以及CollagenⅢ的蛋白表达水平,明确金丝桃苷的抗肾纤维化作用;检测小鼠血尿素氮(BUN)以及血肌酐(Cr)水平,探究金丝桃苷的肾保护作用;使用HE及Masson染色法评价金丝桃苷对小鼠肾组织病理学变化及胶原沉积的影响;Western blot法检测UUO小鼠肾组织以及TGF-β1诱导的NRK-52E细胞中YAP及其下游靶基因CTGF的蛋白表达水平;使用YAP抑制剂维替泊芬,进一步探究金丝桃苷改善肾小管间质纤维化作用机制。结果金丝桃苷可显著逆转UUO小鼠肾组织以及TGF-β1诱导的NRK-52E细胞中α-SMA(P<0.01)、Fibronectin(P<0.01)、CollagenⅠ(P<0.01)以及CollagenⅢ(P<0.01)的蛋白表达水平,同时显著降低UUO小鼠血Cr(P<0.01)、BUN(P<0.01)水平,明显改善肾组织病理学变化以及肾间质胶原沉积;金丝桃苷可在体内外显著抑制TGF-β1诱导的NRK-52E细胞中YAP(P<0.01)、CTGF(P<0.01)蛋白表达水平,且与维替泊芬作用相似,二者均明显改善TGF-β1诱导的NRK-52E细胞纤维化。结论金丝桃苷通过抑制YAP发挥抗肾小管间质纤维化作用。Objective To clarify the anti-fibrotic effect of hypericin on preventing renal fibrosis.To explore if hypericin can improve renal tubulointerstitial fibrosis by inhibiting the Yes-associated protein(YAP).Methods Thirty-six male mice were randomly allocated into 6 groups(n=6 per group):sham group,sham+hypericin(120 mg/kg)group,UUO model group,hypericin(30,60,120 mg/kg)treatment groups.Except for the sham group,renal interstitial fibrosis was induced in all other groups via unilateral ureteral obstruction(UUO).Mice in the hypericin treatment group received corresponding doses of hypericin suspension via oral gavage,while sham group and UUO model group were administered equivalent volumes of normal saline through the same route for 14 consecutive days.Additionally,an in vitro model was established using NRK-52E cells treated with TGF-β1(10 ng/mL).To evaluate the anti-fibrotic effects of hypericin,protein expression levels ofα-SMA,fibronectin,collagen Ⅰ and collagen Ⅲ were assessed in both the UUO mouse kidney tissues and TGF-β1-stimulated NRK-52E cells using Western blotting.Blood urea nitrogen(BUN)and creatinine(Cr)levels in mice were measured to assess the renal protective effects of hypericin.Histological changes and collagen deposition in the kidneys were observed using hematoxylin-eosin and Masson staining.The expression levels of YAP and its downstream target gene CTGF were also determined using Western blotting.The specific mechanism of hypericin in improving renal interstitial fibrosis was further explored using the YAP inhibitor verteporfin.Results The protein expression levels ofα-SMA,FN,collagen Ⅰ,and collagen Ⅲ in renal tissues of UUO mice and TGF-β1-induced NRK-52E cells were markedly reversed by hypericin(all P<0.01).The levels of Cr and BUN in UUO mice serum were effectively reduced(both P<0.01),and renal histopathological alterations and interstitial collagen deposition were markedly ameliorated.Potent inhibitory effects on YAP and CTGF protein expression were observed both in vivo and i

关 键 词：金丝桃苷 肾间质纤维化 肾保护 单侧输尿管梗阻 YAP 

分 类 号：R285[医药卫生—中药学] R692[医药卫生—中医学]