Atractylodis rhizoma water extract attenuates fructose-induced glomerular injury in rats through anti-oxidation to inhibit TRPC6/p-CaMK4 signaling
Atractylodis rhizoma water extract attenuates fructose-induced glomerular injury in rats through anti-oxidation to inhibit TRPC6/p-CaMK4 signaling
复制标题
白术水提取物通过抗氧化抑制 TRPC6/p-CaMK4 信号传导减轻果糖诱导的大鼠肾小球损伤
DOI：
10.1016/j.phymed.2021.153643
复制
发表时间：
2021-07-26
期刊：
影响因子：
7.9
通讯作者：
Kong, Ling-Dong
中科院分区：
文献类型：
--
作者：
Chen, Li;Yang, Jie;Kong, Ling-Dong
背景：白术是一种祛湿芳香草药，在中医中用于治疗肾性水肿已有数千年历史。这种草药具有抗氧化作用。然而，目前尚不清楚苍术如何通过其抗氧化作用来预防肾小球损伤。 目的：通过对苍术水提取物（ARE）成分和网络药理学的分析，探讨ARE是否通过其抗氧化作用抑制氧化应激驱动的瞬时受体电位通道6（TRPC6）及其下游分子钙/钙调蛋白依赖性蛋白激酶IV（CaMK4）信号传导来预防肾小球损伤。方法：液体使用色谱-串联质谱 (LC-MS/MS) 分析 ARE 成分。初步进行了网络药理学分析。雄性 Sprague-Dawley 大鼠给予 10% 果糖饮用水（100 mL/d），持续 16 周。在过去 8 周内，大鼠口服 ARE 剂量为 720 和 1090 mg/kg。分别检测大鼠肾皮质过氧化氢（H2O2）和丙二醛（MDA）水平以及超氧化物歧化酶（SOD）活性。通过蛋白质印迹法分析大鼠肾小球中氧化还原相关因子叉头框 O3 (FoxO3)、SOD2 和过氧化氢酶 (CAT)、足细胞裂孔隔膜蛋白 podocin 和 nephrin、细胞骨架蛋白 CD2 相关蛋白 (CD2AP) 和 α-Actinin-4，以及 TRPC6、p-CaMK4 和突触足蛋白水平。采用qRT-PCR检测SOD2和CAT mRNA水平。结果：ARE中鉴定出36种成分。其中，网络药理学分析表明ARE可能抑制肾脏氧化应激。因此，ARE 上调了果糖喂养大鼠肾小球中 FoxO3 的表达，然后增加了肾小球中 SOD2 和 CAT 的 mRNA 和蛋白质水平。它降低了果糖喂养大鼠肾皮质中的 H2O2 和 MDA 水平，并增加了 SOD 活性。随后，ARE 下调果糖喂养大鼠肾小球中的 TRPC6 和 p-CaMK4，并上调突触足蛋白。此外，ARE 增加了 podocin 和 nephrin，以及 CD2AP 和 α-Actinin-4，这与该动物模型中尿白蛋白与肌酐比值的降低和肾小球结构损伤的改善相一致。结论：这些结果表明，ARE 可能通过减少氧化应激来抑制 TRPC6/p-CaMK4 信号传导并上调突触蛋白表达来预防果糖喂养大鼠的肾小球损伤。因此，ARE可能成为临床上治疗高果糖所致肾小球损伤的一种有前景的药物。
Background: Atractylodis rhizoma, an aromatic herb for resolving dampness, is used to treat Kidney-related edema in traditional Chinese medicine for thousands years. This herb possesses antioxidant effect. However, it is not yet clear how Atractylodis rhizoma prevents glomerular injury through its anti-oxidation.Purpose: Based the analysis of Atractylodis rhizoma water extract (ARE) components and network pharmacology, this study was to explore whether ARE prevented glomerular injury via its anti-oxidation to inhibit oxidative stress-driven transient receptor potential channel 6 (TRPC6) and its downstream molecule calcium/calmodulin-dependent protein kinase IV (CaMK4) signaling.Methods: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to analyze ARE components. Network pharmacology analysis was preliminarily performed. Male Sprague-Dawley rats were given 10% fructose drinking water (100 mL/d) for 16 weeks. ARE at 720 and 1090 mg/kg was orally administered to rats for the last 8 weeks. Hydrogen peroxide (H2O2) and malondialdehyde (MDA) level, and superoxide dismutase (SOD) activity in rat kidney cortex were detected, respectively. In rat glomeruli, redox-related factors forkhead box O3 (FoxO3), SOD2 and catalase (CAT), podocyte slit diaphragm proteins podocin and nephrin, cytoskeleton proteins CD2-associated protein (CD2AP) and alpha-Actinin-4, as well as TRPC6, p-CaMK4 and synaptopodin protein levels were analyzed by Western Blotting. SOD2 and CAT mRNA levels were detected by qRT-PCR.Results: 36 components were identified in ARE. Among them, network pharmacology analysis indicated that ARE might inhibit kidney oxidative stress. Accordingly, ARE up-regulated nuclear FoxO3 expression, and then increased SOD2 and CAT at mRNA and protein levels in glomeruli of fructose-fed rats. It reduced H2O2 and MDA levels, and increased SOD activity in renal cortex of fructose-fed rats. Subsequently, ARE down-regulated TRPC6 and p-CaMK4, and up-regulated synaptopodin in glomeruli of fructose-fed rats. Furthermore, ARE increased podocin and nephrin, as well as CD2AP and alpha-Actinin-4, being consistent with its reduction of urine albumin-to-creatinine ratio and improvement of glomerular structure injury in this animal model.Conclusions: These results suggest that ARE may prevent glomerular injury in fructose-fed rats possibly by reducing oxidative stress to inhibit TRPC6/p-CaMK4 signaling and up-regulate synaptopodin expression. Therefore, ARE may be a promising drug for treating high fructose-induced glomerular injury in clinic.