消脂汤对非酒精性脂肪性肝炎小鼠肠道主要菌群的影响

Effect of Xiaozhitang on Intestinal Flora of Mice with NAFLD

目的:观察消脂汤对非酒精性脂肪性肝炎(NAFLD)小鼠模型的治疗作用及对肠道主要菌群的影响。方法:采用D12492高脂饲料诱导建立NAFLD小鼠模型,造模成功后分别给予不同剂量药物干预,检测血清肿瘤坏死因子-α(TNF-α),白细胞介素-6(IL-6),内毒素(LPS)及相关生化指标,取肝脏行苏木素-伊红(HE)染色和油红O染色;实时荧光定量PCR(Realtime PCR)检测结肠内容物中普雷沃氏菌属、梭杆菌属、双歧杆菌、乳酸杆菌、大肠埃希菌等主要肠道菌群数量的变化。结果:与正常组比较,模型组血清甘油三酯(TG),总胆固醇(TC),血清丙氨酸氨基转移酶(ALT),天冬氨酸氨基转移酶(AST),空腹血糖(FBG),空腹胰岛素(FINS),胰岛素抵抗指数(HOMA-IR),LPS,TNF-α,IL-6水平明显升高(P<0.05,P<0.01);与模型组比较,治疗组FBG,FINS,HOMA-IR,TC,TG显著降低(P<0.01),非诺贝特组血清ALT,AST无明显降低;与正常组比较,模型组小鼠结肠内梭杆菌属、普雷沃氏菌属、双歧杆菌及乳酸杆菌数量明显减少(P<0.01),大肠埃希菌数量升高(P<0.01);正常组与正消组比较无明显差异;与模型组比较,正消组,消脂汤高、中剂量组梭杆菌属、普雷沃氏菌属、双歧杆菌及乳酸杆菌数量明显升高(P<0.05,P<0.01),大肠埃希菌数量显著减少(P<0.01);消脂汤低剂量组、非诺贝特组与模型组无明显差异。结论:消脂汤能显著改善NAFLD小鼠的胰岛素抵抗,减轻肝脏脂肪蓄积和炎症反应,可能与其调整肠道菌群,促进乳酸杆菌、双歧杆菌等优势菌群数量的增长相关。

Objective： To observe the effect of Xiaozhitang on intestinal flora of non-alcoholic fatty liver disease （NAFLD） model. Method： Mice were fed with D12492 high-fat diet for 16 weeks to establish the NAFLD model. After successful modeling, they were randomly divided into model group, high-dose Xiaozhitang group, middle-dose Xiaozhitang group, low-dose Xiaozhitang group, fenofibrate group, normal control group and Xiaozhitang control group. All of the mice were given normal diet. Meanwhile, drug groups were given the corresponding drugs, and control group was given equal volume of normal saline. Nine weeks after administration, all of the mice were treated with intraperitoneal anesthesia with 3% hydrate to collect serum. Total cholesterol（TC） and Triglyceride （TG） content in liver tissues and aspartase aminotransferase （AST） and alanine aminotransferase （ALT） content in serum were detected by automatic biochemical analyzer. The expressions of tumor necrosis factor-alpha （TNF-α） , interleukin-6 （IL-6） and Lpopolysaccharide （LPS） were detected by enzymes linked immunosorbent assay （ELISA）. Two liver tissues were collected for paraffin embedding, hematoxyhin-easin （HE） staining and oil red O staining. Bacterial counts of Prevotella, Bifidobacterium, Escherichia coli, Fusobacterium and Lactobacillus were examined by Real-time polymerase chain reaction （Real- time PCR）. Result： Compared with normal group, blood lipid, insulin （INS） and insulin resistance index （HOMAR-IR） were significantly increased in model group （P 〈 0.05, P 〈 0. 01 ）. Compared with model group, TC, TG, FBG, FINS, HOMA-IR were lower in experimental group （P 〈0.05, P 〈0.01）. However, there was no difference between model group and fenofibrate group. The bacterial counts of Prevotella, Bifidobacterium, Fusobacterium and Lactobacillus in model group were lower than normal group （ P 〈 0.01 ） , and Escherichia coli was higher in model group. The bacterial counts of Prevotella, Bifidobacterium, Fusobacterium and Lactobacillus were much higher in high-dose group and middle-dose group than that of model group （P 〈 0.05, P 〈 0.01 ）. However, the bacterial count of Escherichia coli was lower than model group （P 〈 0.01 ）. There was no difference between model group, the low-dose group and the Fenofibrate group. Conclusion： Xiaozhitang can significantly improve insulin resistance of NAFLD, reduce lipid and inflammation, which may be related to its effects in adjusting intestinal flora and improving the growth of Lactobacillus, Bifidobacterium and other dominant flora.