益生菌联合柴胡疏肝散治疗大鼠非酒精性脂肪肝的实验研究

Rapeutic Effect of Probiotics Combined Chaihu Shugan Powder on Nonalcoholic Fatty Liver Rats

目的观察益生菌联合柴胡疏肝散对非酒精性脂肪肝(NAFLD)大鼠的干预作用,并探讨其可能的作用机制。方法将45只SD大鼠按照随机数字表法分为5组:正常组、模型组、柴胡疏肝散[5 mL/(kg·d)生药]、益生菌组(1 mL/d益生菌)、益生菌联合柴胡疏肝散组[简称联合组,5 mL/(kg·d)生药+1 mL/d益生菌],以高糖高脂饲料诱导SD大鼠制成NAFLD模型,各组分别治疗8周。采用生化试剂盒检测大鼠血清和肝组织中甘油三酯(TG)、总胆固醇(TC)、ALT、AST、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、游离脂肪酸(FFA)水平。ELISA法检测血清中肿瘤坏死因子-α(TNF-α)、IL-18、IL-6、IL-22以及胰岛素(Ins)含量。同时对肝脏组织进行HE染色,观察其组织病理学变化。结果与正常组比较,模型组大鼠TG、TC、ALT、AST、LDL-C、FFA以及炎性因子水平显著升高(P<0.01),HDL-C含量显著降低(P<0.01)。与模型组比较,柴胡疏肝散组、益生菌组、联合组大鼠TG、ALT、AST、LDL-C、FFA、TNF-α、IL-18、IL-6、IL-22及Ins含量降低(P<0.05,P<0.01),HDL-含量显著升高(P<0.01);且联合组效果优于柴胡舒肝散组或益生菌组(P<0.01)。病理结果显示,模型组可见肝细胞排列紊乱,细胞边界不清晰,胞浆中较多脂肪空泡,柴胡舒肝散与益生菌治疗组肝细胞边界逐渐清晰,且脂肪空泡数量减少,联合组肝细胞排列紧密,无明显可见脂肪空泡,接近于正常组水平。结论柴胡疏肝散、益生菌均对NAFLD有一定治疗作用,两者联合有协同作用,能发挥更好的治疗效果,其机制可能与减轻肝脏炎性水平,并调节肝脏脂质代谢有关。

Objective To observe the therapeutic effect of probiotics combined Chaihu Shugan Powder(CHSGP) on nonalcoholic fatty liver disease(NAFLD), and to systematically elucidate its possible mechanism. Methods Totally 45 SD rats were divided into 5 groups by random digit table, i.e., normal, model, CHSGP [5 mL/(kg·d) crude drugs], probiotics(1 mL/d probiotics), probiotics combined CHSGP [combination group, 5 mL/(kg·d) crude drugs+1 mL/d probiotics]. NAFLD model was induced in SD rats by high glucose and high fat diet. Rats in each group were respectively treated for 8 weeks. The levels of triglyceride(TG), total cholesterol(TC), glutamic-pyruvic transaminase(ALT), glutamicoxalacetic transaminase(AST), high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C), and free fatty acid(FFA) in serum and liver tissues were measured by biochemical kits. The serum levels of tumor necrosis factor-α(TNF-α), interleukin-18(IL-18), IL-6, IL-22, and insulin(Ins) were measured by ELISA. At the same time HE staining was performed to observe the histopathological changes of liver tissues. Results Compared with the normal group, the levels of TG, TC, ALT, AST, LDL-C, FFA, and inflammatory factors significantly increased(P<0.01), while the HDL-C content significantly decreased(P<0.01) in the model group. Compared with the model group, the levels of TG, ALT, AST, LDL-C, FFA, TNF-α, IL-18, IL-6, IL-22, and Ins decreased(P<0.05, P<0.01), while the HDL-C content significantly increased(P<0.01) in CHSGP group, probiotics group, and combination group. And the effects in the combination group were better than those of CHSGP group or probiotic group(P<0.01). The pathological results showed that disarranged hepatocytes with unclear boundary could be seen in the model group. There were more fat vacuoles in the cytoplasm. The boundary of hepatocytes in CHSGP group and probiotics group were gradually clear, and the number of fat vacuoles was reduced. The hepatocytes were closely arranged with no obviously visible vacuoles in the combination group, which were close to the levels of the normal group. Conclusions Both CHSGP and probiotics had certain therapeutic effects on NAFLD. Their combinations synergistically exerted better therapeutic effects. The mechanism might be related to attenuating inflammatory levels of the liver and regulating lipid metabolism of the liver.