杨梅素通过激活Nrf2/HO-1通路抑制氧化应激改善肾脏纤维化  

Myricetin attenuates renal fibrosis by activating Nrf2/HO-1pathway to inhibit oxidative stress

作　　者：李冬雪 黄周 王瀚宇 张之昊 谭宁华[1] 邓雪阳 LI Dong-xue;HUANG Zhou;WANG Han-yu;ZHANG Zhi-hao;TAN Ning-hua;DENG Xue-yang(China Pharmaceutical University,School of Traditional Chinese Pharmacy,Nanjing 211198,China)

机构地区：[1]中国药科大学中药学院,江苏南京211198

出　　处：《药学学报》2024年第2期359-367,共9页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金青年资助项目(8200020019)。

摘　　要：本文探讨杨梅素(myricetin,MYR)对单侧输尿管结扎(unilateral uretera obstruction,UUO)和胆管结扎(common bile duct ligation,CBDL)诱导的小鼠肾脏纤维化的影响及其作用机制。动物实验已获得中国药科大学伦理委员会批准,项目伦理号为2022-10-020。选取35只ICR小鼠,分为control组、UUO组、UUO+MYR组、CBDL组、CBDL+MYR组。H&E和Masson染色检测肾脏组织病理变化,蛋白质免疫印迹法(Western blot,WB)检测肾组织中纤维化相关蛋白的表达;总超氧化物歧化酶(superoxide dismutase,SOD)活性检测试剂盒(WST-8法)检测CBDL小鼠肾脏组织中的总SOD变化。体外实验采用HK-2细胞,细胞转化生长因子-β1(transforming growth factor beta 1,TGF-β1)(10 ng·mL^(-1))造纤维化模型,高糖(30 mmol·L-1)造氧化应激模型,随后使用不同浓度的MYR处理,WB检测纤维化及氧化应激相关蛋白的表达;使用不同浓度的MYR处理NIH/3T3细胞,5-溴-2′-脱氧尿苷(5-bromo-2′-deoxyuridine,Brdu)标记法检测其对细胞增殖的影响。结果显示,UUO组和CBDL组的肾脏病变严重,胶原沉积明显,I型胶原蛋白(collagen-I,COL-I)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、纤连蛋白(fibronectin,FN)、波动蛋白(vimentin)、纤溶酶原激活物抑制物1(plasminogen activator inhibitor-1,PAI-1)蛋白表达上调,CBDL组小鼠肾脏SOD酶活力明显下降,MYR给药治疗后逆转了上述变化。MYR给药抑制NIH/3T3细胞的增殖活力而对HK-2细胞没有影响,降低了TGF-β1诱导HK-2细胞后PAI-1、FN、vimentin的上调。MYR给药也能上调高糖诱导HK-2细胞后核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)、血红素加氧酶1(heme oxygenase-1,HO-1)的下调。综上所述,MYR在体内外都可发挥改善肾脏纤维化的作用,可能是通过抑制成纤维细胞的增殖,同时激活Nrf2/HO-1信号通路抑制氧化应激发挥作用。This paper investigates the effect of myricetin(MYR)on renal fibrosis induced by unilateral ureteral obstruction(UUO)and common bile duct ligation(CBDL)in mice and its mechanism.The animal experiment has been approved by the Ethics Committee of China Pharmaceutical University(NO:2022-10-020).Thirty-five ICR mice were divided into control,UUO,UUO+MYR,CBDL and CBDL+MYR groups.H&E and Masson staining were used to detect pathological changes in kidney tissues.Western blot(WB)was used to detect the expression of fibrosis-related proteins in renal tissue,and total superoxide dismutase(SOD)activity detection kit(WST-8)was used to detect the changes of total SOD in renal tissue of CBDL mice.In vitro,HK-2 cells and transforming growth factor beta 1(TGF-β1,10 ng·mL^(-1))were used to induce fibrotic model,and high glucose(30 mmol·L-1)was used to induce oxidative stress model,and then treated with different concentrations of MYR,WB was used to detect the expression of fibrosis and oxidative stress-related proteins,while NIH/3T3 cells were treated with different concentrations of MYR,and their effects on cell proliferation were detected by 5-bromo-2′-deoxyuridine(Brdu).The results showed that the renal lesions in UUO group and CBDL group were severe,collagen deposition was obvious,the expression of collagen-Ⅰ(COL-Ⅰ),α-smooth muscle actin(α-SMA),fibronectin(FN),vimentin and plasminogen activator inhibitor-1(PAI-1)protein was up-regulated,and the activity of SOD enzyme in CBDL group was significantly decreased.MYR partly reversed the above changes after treatment.MYR inhibited the proliferation of NIH/3T3 cells but had no effect on the proliferation of HK-2 cells,and decreased the upregulation of PAI-1,FN and vimentin in HK-2 cells stimulated by TGF-β1.MYR can also upregulate the down-regulation of nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxygenase-1(HO-1)in HK-2 cells stimulated by high glucose.To sum up,MYR can improve renal fibrosis in vivo and in vitro,probably by inhibiting the proliferation of

关 键 词：杨梅素 单侧输尿管结扎 胆管结扎 肾纤维化 氧化应激 

分 类 号：R966[医药卫生—药理学]