Dihydroartemisinin suppresses renal fibrosis in mice by inhibiting DNA-methyltransferase 1 and increasing Klotho  

作　　者：Wei Zhou Min-min Chen Hui-ling Liu Zi-lin Si Wen-hui Wu Hong Jiang Lin-xiao Wang Nosratola D.Vaziri Xiao-fei An Ke Su Cheng Chen Ning-hua Tan Zhi-hao Zhang 

机构地区：[1]State Key Laboratory of Natural Medicines,Department of TCMs Pharmaceuticals,School of Traditional Chinese Pharmacy,China Pharmaceutical University,Nanjing 210009,China [2]Division of Nephrology and Hypertension,School of Medicine,University of California Irvine,Irvine,CA,USA [3]Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210023,China [4]Department of Nephrology,Renmin Hospital of Wuhan University,Wuhan 430060,China

出　　处：《Acta Pharmacologica Sinica》2022年第10期2609-2623,共15页中国药理学报（英文版）

基　　金：This work was financially supported by the National Natural Science Foundation of China(Grants 81703691,81803694);the Fundamental Research Funds for the Central Universities(Grant No.2632020ZD07).

摘　　要：Renal fibrosis is an unavoidable end result of all forms of progressive chronic kidney diseases(CKD).Discovery of efficacious drugs against renal fibrosis is in crucial need.In a preliminary study we found that a derivative of artemisinin,dihydroartemisinin(DHA),exerted strong renoprotection,and reversed renal fibrosis in adenine-induced CKD mouse model.In this study we investigated the anti-fibrotic mechanisms of DHA,particularly its specific target in renal cells.Renal fibrosis was induced in mice by unilateral ureteral obstruction(UUO)or oral administration of adenine(80 mg·kg^(-1)),the mice received DHA(30 mg·kg^(-1)·d^(-1),i.g.)for 14 or 21 days,respectively.We showed that DHA administration markedly attenuated the inflammation and fibrotic responses in the kidneys and significantly improved the renal function in both the renal fibrosis mouse models.In adenine-treated mice,DHA was more effective than 5-azacytidine against renal fibrosis.The anti-fibrotic effects of DHA were also observed in TGF-β1-treated HK-2 cells.In order to determine the target protein of DHA,we conducted pull-down technology coupled with shotgun proteomics using a small-molecule probe based on the structure of DHA(biotin-DHA).As a results,DNA methyltransferase 1(DNMT1)was identified as the anti-fibrotic target of DHA in 3 different types of renal cell lines(HK-2,HEK293 and 3T3).We demonstrated that DHA directly bound to Asn 1529 and Thr 1528 of DNMT1 with a Kd value of 8.18μM.In primary mouse renal tubular cells,we showed that DHA(10μM)promoted DNMT1 degradation via the ubiquitin–proteasome pathway.DHA-reduced DNMT1 expression effectively reversed Klotho promoter hypermethylation,which led to the reversal of Klotho protein loss in the kidney of UUO mice.This subsequently resulted in inhibition of the Wnt/β-catenin and TGF-β/Smad signaling pathways and consequently conferred renoprotection in the animals.Knockdown of Klotho abolished the renoprotective effect of DHA in UUO mice.Our study reveals a novel pharmacological activity

关 键 词：renal fibrosis DIHYDROARTEMISININ DNMT1 KLOTHO 5-AZACYTIDINE affinity-based proteome profiling 

分 类 号：R734.2[医药卫生—肿瘤]