鳖甲煎丸改善肠道菌群失调治疗大鼠非酒精性脂肪肝的机制探讨

Biejia Jianwan Improves Intestinal Flora Imbalance and Attenuates Nonalcoholic Fatty Liver Disease in Rats

摘要:

目的：探讨鳖甲煎丸通过改善肠道菌群失调，进而发挥对非酒精性脂肪肝（non-alcoholic fatty liver disease，NAFLD）模型大鼠的肝脏保护作用。方法：SD大鼠随机分为正常组、模型组、罗格列酮（3 mg·kg^-1）组及鳖甲煎丸（2.4，1.2，0.6 g·kg^-1）组，每组10只。给予高脂乳剂及四氯化碳橄榄油溶液诱导NAFLD模型，造模成功后连续给药4周。在实验过程中动态监测各组大鼠肠道菌群变化；实验结束后取结肠组织，采用实时荧光定量PCR检测各组大鼠结肠紧密连接蛋白（occludin）和闭锁连接蛋白1（ZO-1）mRNA的表达，用阿利新蓝-过碘酸雪夫（AB-PAS）染色考察结肠组织杯状细胞黏液层厚度的变化；同时动态监测大鼠血清丙氨酸氨基转移酶（ALT），天冬氨酸转移酶（AST），碱性磷酸酶（ALP），总胆固醇（TC），低密度脂蛋白-胆固醇（LDL-C）指标，并用苏木精-伊红（HE）染色观察肝脏组织病变情况。结果：与正常组比较，模型组拟杆菌门、拟杆菌属、梭杆菌属数量明显增加（P<0.05），乳酸杆菌数量减少（P<0.05），肠道紧密连接蛋白occludin表达明显减少（P<0.05），肠道通透性明显增加（P<0.05）。与模型组比较，鳖甲煎丸（0.6 g·kg^-1）组拟杆菌1和拟杆菌2数量明显减少（P<0.05），同时乳酸杆菌数量增加（P<0.05），结肠occludin表达明显增加（P<0.05），同时有效降低肝损伤指标，炎性细胞浸润明显改善。结论：鳖甲煎丸能够改善NAFLD模型大鼠肠道菌群紊乱，有效降低肠道通透性，显著降低肝细胞损伤。

Abstract:

Objective: To study the effect of Biejia Jianwan on intestinal flora imbalance and investigate its protection effect on liver in non-alcoholic fatty liver disease (NAFLD) rats model. Method: SD rats were randomly divided into normal group, model group, rosiglitazone (3 mg·kg^-1) group and Biejia Jianwan (2.4, 1.2 and 0.6 g·kg^-1) groups (n=10 in each group). NAFLD rat models were established by high fat emulsion and carbon tetrachloride olive oil solution. The drugs were given for 4 weeks after successful modeling. The changes in intestinal flora were monitored dynamically. Colon tissues were taken after experiment to detect mRNA expression levels of tight junction protein expression occludin and blocking junction protein (ZO-1) with the method of Real-time PCR. In addition, mucus layer thickness was evaluated by Alcian blue-periodic acid Schiff (AB-PAS) staining. The levels of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), total cholesterol (TC), and low density lipoprotein cholesterol (LDL-C) in blood serum were monitored dynamically. The liver histopathology was observed by means of hematoxylin-eosin (HE) staining. Result: As compared with the normal group, the number of Bacteroidetes, Bacteroides and Fusobacterium was significantly increased (P<0.05) and the number of Lactobacilli was decreased in model group (P<0.05). Besides, the expression level of occludin mRNA was significantly decreased and intestinal permeability was significantly increased (P<0.05) in model group. As compared with the model group, Biejia Jianwan 0.6 g·kg^-1 could significantly reduce the number of Bacteroidetes, while increase the number of Lactobacilli and the expression level of occludin mRNA (P<0.05). Biejia Jianwan could also effectively relieve liver damage and improve inflammatory cell infiltration. Conclusion: Biejia Jianwan had protective effects in NAFLD model rats by improving intestinal flora disturbance, effectively reducing intestinal permeability and significantly reducing liver damage.