槲皮素通过调控内质网应激信号通路改善非酒精性脂肪肝大鼠肝脏损伤
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罗瑞熙,E-mail:luoruixi058@gzy.edu.cn

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国家自然科学基金(82060824);贵州省科学技术基金(ZK[2021]YB345);贵州省卫生健康委员会科学技术基金 (2021[92]);贵州省教育厅高等学校科学研究项目(青年项目)(2022[203])


Quercetin attenuates liver injury in rats with nonalcoholic fatty liver disease by modulating endoplasmic reticulum stress
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    摘要:

    目的:基于内质网应激(endoplasmic reticulum stress,ERS)信号通路探讨槲皮素(quercetin,Que)对高脂饲料诱导的非酒精性脂肪肝(non-alcoholic fatty liver disease,NAFLD)大鼠模型的干预作用及其机制。方法:高脂饲料喂养SD大鼠8周建立NAFLD模型,之后随机分为模型组、Que低剂量组(25 mg/kg)和Que高剂量组(50 mg/kg),另设正常饮食大鼠作为正常对照组,给药期间监测大鼠体重、进食量。继续饲养8周后,检测各组大鼠血清中胆固醇(total cholesterol,TC)、甘油三酯(triglyceride, TG)、低密度脂蛋白胆固醇(low-density lipoprotein-cholesterol,LDL-C)、高密度脂蛋白胆固醇(high-density lipoprotein-cholesterol, HDL-C)、丙氨酸氨基转移酶(glutamic-pyruvic transaminase,ALT)、天冬氨酸氨基转移酶(glutamic oxalacetic transaminase,AST) 以及肝脏组织中TC、TG的含量变化;苏木素-伊红(hematoxylin-eosin,HE)染色、油红O染色、马松(Masson)染色观察大鼠肝脏组织的病理学变化;血糖仪检测大鼠血糖值并通过糖耐量试验评估大鼠糖耐量受损情况;酶联免疫吸附试验(enzyme-linked immuno sorbent assay,ELISA)试剂盒检测各组大鼠空腹胰岛素水平;实时荧光定量PCR法(quantitative real-time PCR,RT-qPCR)检测各组大鼠肝脏组织ERS相关基因 BipAtf6Atf4Xbp-1sChop 的表达变化;Western blot 法检测各组大鼠肝脏组织 ERS 相关蛋白 BiP、CHOP、ATF4、ATF6、XBP-1s和XBP-1u的表达变化。结果:实验期间,各组大鼠体重、进食量差异无统计学意义;与正常对照组比较,模型组大鼠血清TG、TC、LDL-C、AST、ALT浓度升高,HDL-C浓度降低(P < 0.05);与模型组比较,Que低剂量组大鼠血清TG、TC、LDL-C、AST、ALT浓度降低(P < 0.05),Que高剂量组大鼠血清TG、TC、AST、ALT浓度降低(P < 0.05)。与正常对照组比较,模型组大鼠肝脏体积增大,重量增加,肝细胞形态明显肿胀且呈空泡化,脂质沉积明显增加,且纤维化区域明显增加,同时大鼠出现胰岛素抵抗和糖耐量受损情况;与模型组比较,以上病理改变在 Que 低、高剂量组大鼠中有较明显改善。RT-qPCR和Western blot结果显示,与正常对照组比较,模型组大鼠肝脏组织中BipAtf6Atf4Xbp-1sChop mRNA表达显著升高,BiP、CHOP、ATF4、ATF6和XBP-1s蛋白表达显著升高;与模型组比较,Que低、高剂量组大鼠肝脏组织BipAtf6Atf4Xbp-1sChop mRNA 表达降低,BiP、CHOP、ATF4、ATF6和XBP-1s蛋白表达也显著降低。结论:Que可以改善高脂饲料诱导的 NAFLD大鼠脂代谢异常、糖耐量受损、肝脏脂质沉积以及病理损伤,其机制可能是通过调控ERS信号通路发挥作用。

    Abstract:

    Objective:This study aimed to investigate the potential of quercetin(Que)in ameliorating non-alcoholic fatty liver disease (NAFLD)induced by a high-fat diet(HFD)through the modulation of endoplasmic reticulum stress(ERS)in rats. Methods: Feeding SD rats with high-fat diet(HFD)for 8 weeks to establish the NAFLD model,then randomly divided into the HFD group,HFD + low-dose Que group(25 mg/kg),and HFD + high-dose Que group(50 mg/kg). Another group of rats fed normal diet served as the control group. During the administration period,monitoring of rat body weight and food intake was conducted. After continuing feeding for 8 weeks,serum levels of total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),alanine aminotransferase(ALT),aspartate aminotransferase(AST),as well as changes in TC and TG content in liver tissue of each group were detected. Histopathological changes in rat liver tissue were observed by hematoxylin -eosin (HE)staining,oil red O staining,and Masson’s staining. Blood glucose levels were measured using a glucometer,and rat glucose tolerance was assessed by glucose tolerance test. Fasting insulin levels in each group of rats were detected using enzyme-linked immunosorbent assay(ELISA)kits. Real-time fluorescent quantitative PCR was used to detect the expression changes of ERS-related genes BipAtf6Atf4Xbp-1s,and Chop in rat liver tissue of each group. Western blot was used to detect the expression changes of ERS-related proteins BiP,CHOP,ATF4,ATF6,XBP-1s,and XBP-1u in rat liver tissue of each group. Results:Throughout the experiment, no significant differences in food intake and body weight were observed among the four groups. Compared to the control group,the HFD group exhibited increased serum levels of triglycerides(TG),total cholesterol(TC),low-density lipoprotein-cholesterol(LDL-C), glutamic oxalacetic transaminase(AST),and glutamic-pyruvic transaminase(ALT),along with decreased high-density lipoprotein-cholesterol(HDL-C)levels(P < 0.05). In the low-dose Que group,serum levels of TG,TC,LDL-C,AST,and ALT were significantly reduced compared to the HFD group(P < 0.05). Similarly,the high-dose Que group showed decreased serum levels of TG,TC,AST, and ALT compared to the HFD group(P < 0.05). In the HFD group,the liver exhibited enlargement and increased absolute weight, with prominently swollen hepatocytes,marked vacuolization,and accumulated lipid droplets. Additionally,increased collagen deposition,insulin resistance,and impaired glucose tolerance were observed in the HFD group. However,these alterations were reversed in both low-and high-dose Que groups. The results of RT-qPCR and Western blot showed that compared to the control group, the mRNA expression of BipAtf6Atf4Xbp-1s,and Chop in the liver tissue of rats in the HFD group significantly increased,and the protein expression of BiP,CHOP,ATF4,ATF6,and XBP-1s also significantly increased. Compared to the HFD group,the mRNA expression of BipAtf6Atf4Xbp-1s,and Chop in the liver tissue of rats in the low-and high-dose Que groups decreased,and the protein expression of BiP,CHOP,ATF4,ATF6,and XBP -1s also significantly decreased. Conclusion:Que treatment attenuates HFD-induced aberrant lipid metabolism,impaired glucose tolerance,and hepatic lipid deposition in NAFLD rats,with modulation of the ERS-related pathway potentially playing a critical role.

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罗瑞熙,王文佳,王平,田维毅.槲皮素通过调控内质网应激信号通路改善非酒精性脂肪肝大鼠肝脏损伤[J].南京医科大学学报(自然科学版),2024,(4):445-454

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  • 收稿日期:2023-07-16
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  • 在线发布日期: 2024-04-11
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