Elabela通过调节水通道蛋白2改善糖尿病肾病作用及机制研究  

Elabela alleviates diabetic kidney disease by regulating aquaporin 2

作　　者：石敏[1] 徐玉迪 王子葳 宋婧 张红[1] Shi Min;Xu Yudi;Wang Ziwei;Song Jin;Zhang Hong(Department of Endocrinology,The Affiliated Huai′an No.1 People′s Hospital of Nanjing Medical University,Huai′an 223300,China)

机构地区：[1]南京医科大学附属淮安第一医院内分泌科,223300

出　　处：《中华内分泌代谢杂志》2022年第8期651-657,共7页Chinese Journal of Endocrinology and Metabolism

基　　金：国家自然科学基金(81700723);江苏省自然科学基金(BK20191213)。

摘　　要：目的观察Elabela(Ela)对糖尿病肾病(DKD)保护作用及可能机制。方法db/db小鼠随机分为糖尿病组和Ela干预组,db/m小鼠为正常对照组。Ela干预组小鼠腹腔注射Ela-215 mg·kg^(-1)·d^(-1),持续8周。实验结束测尿白蛋白/肌酐比值(UACR)等代谢指标,HE染色和PAS染色观察肾脏病理学改变,免疫组化观察水通道蛋白2(AQP2)表达,Western blot分析肾脏Ⅳ型胶原蛋白(Col-Ⅳ)和AQP2水平。Ela处理高糖诱导人肾小管上皮细胞(HK-2),Western blot观察HK2细胞外基质主要成分Col-Ⅳ及AQP2的表达;进一步敲除HK2细胞apelin受体(APJ),观察Ela的干预效应。最后,利用NanoBit?技术,研究Ela诱导的APJ激活与抗利尿激素(AVP)诱导的抗利尿激素2型受体(AVPR2)活化间的相互作用,以明确Ela调节AQP2的可能机制。结果(1)在不影响血糖、体重等代谢指标的情况下,Ela干预明显降低了db/db小鼠的UACR水平,且肾脏病理损害显著改善,肾组织Col-Ⅳ水平下降,AQP2表达明显降低。(2)Ela干预可明显抑制高糖诱导的Col-Ⅳ表达和AQP2水平,进一步干扰APJ后,Ela对Col-Ⅳ和AQP2调节作用被逆转。(3)Ela激活APJ可明显拮抗AVP诱导的AVPR2下游信号β-Arrestin-2(ARRB2)传递,进一步提示Ela对AQP2的抑制作用可能与拮抗AVP/AVPR2信号有关。结论Ela可能通过APJ介导作用,抑制AQP2水平,发挥DKD保护作用。Objective To assess the renal-protective effect of Elabela(Ela)on diabetic kidney disease(DKD),and explore its potential mechanism.Methods db/db mice were randomly divided into diabetic group and Ela intervention group,while db/m mice were taken as normal control group.The mice in the Ela intervention group were intraperitoneally injected with Ela-215 mg·kg^(-1)·d^(-1)for 8 weeks.At the end of the experiment,urinary albumin/creatinine ratio(UACR)was measured.The renal pathological changes were observed by HE and PAS staining.The expression of aquaporin 2(AQP2)examined by immunohistochemistry.The level of collageⅣ(Col-Ⅳ)and AQP2 in renal tissue was analyzed by Western blot.The human renal tubular epithelial cells(HK-2)were incubated with high glucose medium and further interfered with apelin receptors(APJ)-siRNA.Western blot analysis was used to detect the effect of Ela intervention on Col-Ⅳand AQP2 expression.Finally,to clarify the possible mechanism of Ela regulating AQP2,the interaction between Ela-induced APJ activation and arginine vasopressin(AVP)-evoked arginine vasopressin receptor 2(AVPR2)activation was investigated by NanoBit©technology.Results(1)Without affecting blood glucose and body weight,Ela intervention significantly reduced the UACR in db/db mice,and attenuate pathological changes of the kidney,as well as expression of Col-Ⅳand AQP2.(2)Ela treatment could remarkably inhibit the high glucose-induced the expression of Col-Ⅳand AQP2,which was reversed by interfering with APJ.(3)AVP-induced downstreamβ-Arrestin-2 signaling transduction via AVPR2 was obviously antagonized by interaction of Ela and APJ,further suggesting that the inhibitory effect of Ela on AQP2 may be related to antagonizing AVP/AVPR2 signaling.Conclusion Ela exerts renal protection by inhibiting the expression of AQP2 through APJ.

关 键 词：Elabela 糖尿病肾病 水通道蛋白2 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]