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目的观察复方楂金颗粒对高脂高果糖饮食诱导的非酒精性脂肪性肝炎(nonalcoholic steatohepatitis,NASH)小鼠肝脏内Toll样受体4(Toll-like receptor 4,TLR4)信号通路影响。方法 6周龄雄性C3H品系小鼠30只,按体重分层随机分为高脂高果糖组(20只)和对照组(10只)。高脂高果糖组用高脂饲料和20%果糖饮用水喂养;对照组用普通饲料和普通饮用水喂养。8周后将高脂高果糖组按体重分层随机分为高脂高果糖组和楂金颗粒组各10只,楂金颗粒组用高脂饲料和20%果糖饮用水喂养,每天按50mL/kg予12.8%的复方楂金颗粒冲剂灌胃(山楂、郁金、泽泻、海藻、象贝、水飞蓟、桃仁按3∶1.5∶1.5∶2∶1.5∶2∶1比例制备而成)。余组喂养如前,每天予同体积蒸馏水灌胃,16周处死小鼠。检测小鼠体重、肝湿重、肝功能及脂代谢指标,采用HE、油红O及Masson染色评价小鼠肝脏病理学改变。用免疫组织化学染色和实时荧光定量PCR法检测小鼠肝组织TLR4、髓样分化因子88(myeloid differentiation factor 88,MyD88)、肿瘤坏死因子(tumor necrosis factor-alpha,TNF-α)表达。结果楂金颗粒组体重、谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)明显低于高脂高果糖组(P<0.05);油红O染色面积、密度较对照组减少;HE染色显示,楂金颗粒组炎症气球样变较高脂高果糖组减少;Masson染色阴性;TLR4、MyD88免疫组化染色阳性率和mRNA表达均明显低于高脂高果糖组(P<0.05)。结论复方楂金颗粒剂能明显抑制NASH小鼠肝脏内TLR4信号通路。
Objective To observe the effect of compound hawthorn gold granules on the Toll-like receptor 4 (TLR4) signaling pathway in the liver of nonalcoholic steatohepatitis (NASH) mice induced by high fat and high fructose diet. Methods Thirty male C3H mice of 6 weeks old were randomly divided into high fat and high fructose group (n = 20) and control group (n = 10) by body weight. The high-fat and high-fructose group was fed with high-fat diet and 20% fructose drinking water while the control group was fed with normal feed and ordinary drinking water. After 8 weeks, the high-fat and high-fructose groups were randomly divided into 10 groups according to the body weight. Each group was fed with high-fat and high-fructose group and hawthorn gold granules group. The hawthorn gold granules group was fed with high fat diet and 20% fructose drinking water, kg to 12.8% Compound Hawthorn gold granules granules (hawthorn, turmeric, alisma, seaweed, asbestos, milk thistle, peach kernel ratio of 3: 1.5: 1.5: 2: 1.5: . The remaining groups were fed as before, with the same volume of distilled water every day, mice were killed at 16 weeks. The body weight, liver wet weight, liver function and lipid metabolism index were detected. The pathological changes of liver in mice were evaluated by HE, oil red O and Masson staining. The expressions of TLR4, myeloid differentiation factor 88 (MyD88) and tumor necrosis factor-alpha (TNF-α) were detected by immunohistochemistry and real-time fluorescence quantitative PCR. Results Body weight, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in hawthorn gold granules group were significantly lower than those in high fat and high fructose group (P <0.05); Oil red O staining area and density were lower than those in control group ; HE staining showed that the inflammatory balloon-like changes in hawthorn gold granules group were lower than those in high-fat and high-fructose group; Masson staining was negative; the positive rate and mRNA expression of TLR4 and MyD88 in immunohistochemical staining were significantly lower than those in high fat and high fructose group ). Conclusion Fufang Jinzhong Granules can significantly inhibit the TLR4 signaling pathway in the liver of NASH mice.