Combined melatonin and poricoic acid A inhibits renal fibrosis through modulating the interaction of Smad3 and β-catenin pathway in AKI-to-CKD continuum
Combined melatonin and poricoic acid A inhibits renal fibrosis through modulating the interaction of Smad3 and β-catenin pathway in AKI-to-CKD continuum
复制标题
褪黑激素和猪苓酸 A 组合通过调节 AKI-to-CKD 连续体中 Smad3 和 β-catenin 通路的相互作用抑制肾纤维化
DOI：
10.1177/2040622319869116
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发表时间：
2019-08-01
影响因子：
3.5
通讯作者：
Zhao, Ying-Yong
中科院分区：
文献类型：
--
作者：
Chen, Dan-Qian;Cao, Gang;Zhao, Ying-Yong
背景：急性肾损伤（阿基）是进展为慢性肾脏病（CKD）和肾纤维化的主要危险因素之一。然而，有效的治疗方法仍然知之甚少。在此，我们研究了从茯苓表层分离的褪黑激素和多孔酸A（PAA）的肾脏保护作用，并研究了联合治疗对肾缺血再灌注损伤（IRI）和缺氧/复氧（H/R）或TGF-β 1诱导的HK-2细胞模型中TGF-β/Smad和Wnt/β-连环蛋白相互作用的影响。方法：采用Western blot和免疫组化染色检测蛋白表达，qRT-PCR检测mRNA表达。采用免疫共沉淀、染色质免疫共沉淀、RNA干扰和荧光素酶报告基因分析等方法探讨PAA和褪黑素的肾脏保护作用机制。结果如下：PAA和联合治疗具有肾脏保护和抗纤维化作用，但在AKI到CKD连续体中的潜在机制不同。褪黑素抑制Smad依赖性和非依赖性通路，PAA通过阻断Smad 3与TGF β RI和SARA的相互作用，选择性抑制Smad 3的磷酸化。进一步研究表明，褪黑素和PAA的抑制作用部分依赖于Smad 3，尤其是PAA。褪黑激素和PAA也抑制Wnt/β-连环蛋白通路及其促纤维化下游靶点，PAA表现更好。我们进一步确定，IRI诱导核Smad 3/beta-catenin复合物，而褪黑素和PAA干扰Smad 3和beta-catenin的相互作用，补充PAA可以增强褪黑素对TGF-β/Smad和Wnt/beta-catenin通路的抑制作用。结论：联合褪黑素和PAA为治疗AKI-CKD连续过程中的肾纤维化提供了一种有前途的治疗策略。
Background: Acute kidney injury (AKI) is one of the major risk factors for progression to chronic kidney disease (CKD) and renal fibrosis. However, effective therapies remain poorly understood. Here, we examined the renoprotective effects of melatonin and poricoic acid A (PAA) isolated from the surface layer of Poria cocos, and investigated the effects of combined therapy on the interaction of TGF-beta/Smad and Wnt/beta-catenin in a rat model of renal ischemia-reperfusion injury (IRI) and hypoxia/reoxygenation (H/R) or TGF-beta 1-induced HK-2 cells. Methods: Western blot and immunohistochemical staining were used to examine protein expression, while qRT-PCR was used to examine mRNA expression. Coimmunoprecipitation, chromatin immunoprecipitation, RNA interference, and luciferase reporter gene analysis were employed to explore the mechanisms of PAA and melatonin's renoprotective effects. Results: PAA and combined therapy exhibited renoprotective and antifibrotic effects, but the underlying mechanisms were different during AKI-to-CKD continuum. Melatonin suppressed Smad-dependent and Smad-independent pathways, while PAA selectively inhibited Smad3 phosphorylation through distrupting the interactions of Smad3 with TGF beta RI and SARA. Further studies demonstrated that the inhibitory effects of melatonin and PAA were partially depended on Smad3, especially PAA. Melatonin and PAA also inhibited the Wnt/beta-catenin pathway and its profibrotic downstream targets, and PAA performed better. We further determined that IRI induced a nuclear Smad3/beta-catenin complex, while melatonin and PAA disturbed the interaction of Smad3 and beta-catenin, and supplementing with PAA could enhance the inhibitory effects of melatonin on the TGF-beta/Smad and Wnt/beta-catenin pathways. Conclusions: Combined melatonin and PAA provides a promising therapeutic strategy to treat renal fibrosis during the AKI-to-CKD continuum.