降脂颗粒通过减轻脂肪组织功能障碍改善小鼠非酒精性脂肪性肝炎

Jiangzhi Granule improves nonalcoholic steatohepatitis in mice through alleviating adipose tissue dysfunction

目的:基于脂肪组织功能障碍研究降脂颗粒改善非酒精性脂肪性肝炎(NASH)的作用及机制。方法:C57BL/6J小鼠给予高脂饲料联合高果糖/葡萄糖饮水HFCD-HF/G诱导10周,建立NASH模型;同时随机取8只正常小鼠,设为对照组,给予普通饲料。造模后,模型小鼠随机分为模型组、罗格列酮组及降脂颗粒高、低剂量组,每组8只。对照组小鼠给予普通饲料;其他组小鼠给予HFCD-HF/G,用药组同时每天灌胃给予相应药物。连续干预8周后,测定小鼠体质量、肝质量、附睾脂肪质量;试剂盒检测血清丙氨酸氨基转移酶(ALT)、总胆固醇(TC)水平;HE染色观察肝脏、脂肪组织病理学变化;流式细胞术检测小鼠附睾脂肪组织中巨噬细胞数量和M1/M2分型比例;RT-qPCR分析附睾脂肪组织炎性细胞因子肿瘤坏死因子-α(TNF-α)、白介素(IL)-6、IL-10、单核细胞趋化蛋白-1(MCP-1)mRNA的表达水平,以及脂质代谢相关基因脂联素(ADIPOQ)、葡萄糖转运蛋白4(GLUT4)、脂肪酸结合蛋白4(FABP4)、磷酸烯醇式丙酮酸羧激酶1(PCK1)的表达水平。结果:①与对照组相比,模型组小鼠体质量、肝质量、附睾脂肪质量均明显增加(P<0.01)。与模型组相比,降脂颗粒各组小鼠体质量明显降低(P<0.05),降脂高剂量组小鼠肝质量明显降低(P<0.05),附睾脂肪质量有降低趋势;降脂颗粒各组小鼠血清ALT、TC水平明显下降(P<0.01)。②模型组出现大面积肝组织脂肪变,伴有肝小叶炎症和大量气球样变;降脂颗粒干预能明显改善这些病理变化,非酒精性脂肪性肝病活动度积分(NAS)评分明显下降(P<0.01)。③与模型组小鼠相比,降脂颗粒高剂量组小鼠附睾脂肪组织中脂肪细胞体积减小,炎细胞浸润减少。④降脂颗粒干预后,小鼠附睾脂肪组织巨噬细胞数量减少(P<0.05),M1/M2型巨噬细胞比例降低(P<0.05);促炎因子IL-6、MCP-1、TNF-α mRNA水平明显下降(P<0.05,P<0.01),抗炎因子IL-10 mRNA水平上升(P<0.05);ADIPOQ、GLUT4 mRNA水平上升(P<0.05),FABP4、PCK1 mRNA水平下降(P<0.05)。结论:降脂颗粒能有效改善小鼠NASH,其可能通过减轻脂肪组织病理及功能障碍发挥治疗NASH的作用。

Objective:To explore the effect and mechanism of Jiangzhi Granule on nonalcoholic steatohepatitis(NASH)based on adipose tissue dysfunction.Methods:C57BL/6J mice were fed with high-fat and calorie diet plus high-fructose/glucose drinking water for 10 weeks to establish NASH mode,while 8 normal mice were randomly selected as the control group and given ordinary diet. After modeling,the model mice were randomly divided into model group,rosiglitazone group,low and high dose Jiangzhi Granule group(JZG-L and JZG-H group),with 8 mice in each group. Mice in the control group were given ordinary diet,mice in the other groups were given highfat diet,and mice in the treating groups were given corresponding drugs by gavage every day. After continuous intervention for 8 weeks,the body weight,liver weight and epididymis fat weight of mice were measured. The serum levels of alanine aminotransferase(ALT)and total cholesterol(TC)were measured by commercial kits. Histopathological changes of liver and adipose tissue were observed by HE staining.The number of macrophages and M1/M2 typing ratio in epididymal adipose tissue of mice were detected by flow cytometry. RT-qPCR was used to analyze the mRNA expression levels of inflammatory cytokines such as tumor necrosis factor(TNF)-α,interleukin(IL)-6,IL-10and monocyte chemotactic protein 1(MCP-1)in epididymal adipose tissue. The expression levels of lipid metabolism related genes including adiponectin(ADIPOQ),glucose transporter 4(GLUT4),fatty acid binding protein 4(FABP4)and phosphoenolpyruvate carboxykinase 1(PCK1)were also analyzed.Results:① Compared with the control group,the body weight,liver weight and epididymal fat weight of mice in the model group significantly increased(P<0.01). Compared with the model group,the body weight of mice in the JZG-L and JZG-H group significantly decreased(P<0.05),the liver weight of mice in the JZG-H group significantly decreased(P<0.05),epididymal fat weight showed a downward trend;the serum levels of ALT and TC of mice in JZG-L and JZG-H groups significantly decreased(P<0.01).②In the model group,extensive hepatic steatosis was found in mice liver tissues with hepatic lobule inflammation and a lot of ballooning degeneration,which were improved by Jiangzhi Granule treatment,and the NAS scores were significantly decreased as well(P<0.01).③Compared with the model group,the size of adipocyte and the inflammatory infiltration in epididymal adipose tissue of mice in JZG-H group reduced.④ After intervention with Jiangzhi Granule,the number of macrophages and the proportion of M1/M2macrophages in mice epididymal adipose tissues decreased(P<0.05);the mRNA levels of pro-inflammatory cytokines IL-6,MCP-1 and TNF-α significantly decreased(P<0.05,P<0.01)and the mRNA level of anti-inflammatory cytokine IL-10 increased(P<0.05);the mRNA levels of ADIPOQ and GLUT4 increased(P<0.05)and the mRNA expressions of FABP4 and PCK1 decreased(P<0.05).Conclusion:Jiangzhi Granule can effectively improve NASH in mice,which may play a role in the treatment of NASH by alleviating the pathology and dysfunction of adipose tissue.