基于PI3K/AKT1通路探讨山西老陈醋对高脂饮食诱导大鼠抗氧化作用机制

申彤; 刘佳佳; 陈嘉; 庞全海

doi:10.13386/j.issn1002-0306.2022110310

基于PI3K/AKT1通路探讨山西老陈醋对高脂饮食诱导大鼠抗氧化作用机制

Explore the Mechanism of Shanxi Aged Vinegar on High-fat Diet-Induced Antioxidant in Rats Based on PI3K/AKT1 Pathway

摘要

摘要: 目的：基于PI3K/AKT1通路探究山西老陈醋对高脂饮食诱导大鼠抗氧化作用机制。方法：30只SPF级雄性Wistar大鼠随机分为5组：正常组（C，常规饲料）、高脂模型组（M，高脂乳剂）、低（LV）、中（MV）、高（HV）剂量山西老陈醋干预组（1.35、2.7、5.4 g/（kg·bw））。造模（5周）同时给予陈醋干预后，山西老陈醋继续干预5周。检测血清血脂含量、肝损伤指标及抗氧化酶活性；HE染色观察肝组织病理学状态；qRT-PCR和Western blotting检测大鼠肝脏中PIK3CA、AKT1 mRNA及蛋白水平表达，免疫组织化学技术进行定位分析。结果：与C组相比，M组肝脂肪变性严重，血清中脂质含量和肝损伤程度极显著升高（P<0.01），抗氧化酶活性极显著下降（P<0.01），肝组织中PIK3CA和AKT1 mRNA及蛋白水平表达极显著升高（P<0.01），高脂血症大鼠造模成功，同时PI3K/AKT1通路被激活。血清学检测结果显示，山西老陈醋可显著下调血清中脂质含量、改善肝损伤、增加抗氧化酶活性（P<0.05），使其肝组织病理损伤减轻，且HV和MV组改善效果优于LV组。qRT-PCR、Western blotting及免疫组织化学技术检测结果显示，PIK3CA、AKT1主要定位于肝细胞质及细胞核中，山西老陈醋可极显著下调PI3K/AKT1通路中PIK3CA、AKT1 mRNA及蛋白水平表达（P<0.01），HV和MV组干预效果优于LV组。结论：不同剂量山西老陈醋可不同程度改善高脂饮食大鼠脂质、抗氧化及肝功能指标，最佳剂量可能在2.7~5.4 g/（kg·bw）之间，山西老陈醋可能通过调控PI3K/AKT1通路发挥抗氧化应激损伤作用，进而改善高脂饮食诱发的大鼠高脂血症。

Abstract: Objective: To explore the antioxidant mechanism of Shanxi aged vinegar on rats induced by high fat diet based on PI3K/AKT1 pathway. Methods: Thirty SPF male Wistar rats were randomly divided into 5 groups: Normal group (C, conventional diet), high fat model group (M, high fat emulsion), low (LV), medium (MV) and high (HV) Shanxi aged vinegar intervention groups (1.35, 2.7, 5.4 g/(kg·bw)). Modeling (5 weeks) at the same time given Shanxi aged vinegar intervention, the aged vinegar intervention continued for 5 weeks. Serum lipid content, liver injury index and antioxidant enzyme activity were detected. The histopathological status of liver was observed by HE staining. The mRNA and protein levels of PIK3CA and AKT1 in rat liver were detected by qRT-PCR and Western blotting, and the localization was analyzed by immunohistochemistry. Results: Compared with group C, liver steatosis was severe in group M, serum lipid content and degree of liver injury were significantly increased (P<0.01), antioxidant enzyme activity was significantly decreased (P<0.01), mRNA and protein expressions of PIK3CA and AKT1 in liver tissue were significantly increased (P<0.01). The hyperlipidemia rats were successfully modeled and PI3K/AKT1 pathway was activated. The results of Serological showed that Shanxi aged vinegar could significantly down-regulate serum lipid content, improve liver injury, increase antioxidant enzyme activity (P<0.05), and reduce pathological injury of liver tissue, and the improvement effect of HV and MV groups was better than that of LV group. The results of qRT-PCR, Western blotting and immunohistochemical detection showed that PIK3CA and AKT1 were mainly located in the cytoplasm and nucleus of liver cells. Shanxi aged vinegar could significantly down-regulate the mRNA and protein expressions of PIK3CA and AKT1 in PI3K/AKT1 pathway (P<0.01), and the intervention effect of HV and MV groups was better than that of LV group. Conclusion: Different doses of Shanxi aged vinegar can improve lipid, antioxidant and liver function indexes of rats with high fat diet to different degrees, and the best dose may be between 2.7~5.4 g/(kg·bw). Shanxi aged vinegar may play a role in anti-oxidative stress injury by regulating PI3K/AKT1 pathway, so as to improve hyperlipidemia induced by high fat diet in rats.