双歧杆菌乳杆菌三联活菌干预NASH模型大鼠代谢性炎症的实验研究

Experimental study on prevention and treatment of metabolic inflammation in NASH model rats by live combined bifidobacterium and Lactobacillus tablets

目的:研究双歧杆菌乳杆菌三联活菌对高脂饮食诱发的非酒精性脂肪性肝炎(NASH)模型大鼠代谢性炎症的干预作用。方法:SPF级雄性SD大鼠32只,随机分为3组,其中正常对照组、益生菌干预组各8只,模型对照组16只。正常对照组大鼠以标准饲料饲养,其余2组大鼠采用高脂饲料饲养。每4周从模型对照组随机选取2只大鼠处死以评估模型的复制情况,经HE染色证实第4周末模型对照组大鼠出现肝脂肪变,于实验第5周,益生菌干预组给予双歧杆菌乳杆菌三联活菌片0.7 g·kg^(-1) d^(-1)灌胃;正常对照组和模型对照组予以等体积的0.9%氯化钠灌胃,共干预12周。末次给药后24 h,采集血清检测血清丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST),肝脏匀浆组织检测甘油三酯(TG)、游离脂肪酸(FFA);鲎试剂显色基质法测定门静脉血浆脂多糖(LPS);ELISA法检测白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平。结果:模型对照组大鼠于实验第16周复制成NASH模型,与模型对照组比较,益生菌干预组大鼠血清ALT、AST和肝脏匀浆中TG、FFA明显降低(P<0.05);益生菌干预可以改善NASH模型大鼠血清TNF-α、IL-1β和血浆LPS水平(P<0.05)。结论:双歧杆菌乳杆菌三联活菌对高脂饲料诱发的大鼠NASH具有一定的防治作用,其作用机制可能与通过降低NASH模型大鼠内毒素血症、改善代谢性炎症相关。

Objective:To study the preventive and therapeutic effects of Live Combined Bifidobacterium and Lactobacillus tablets on metabolic inflammation in rats with non-alcoholic steatohepatitis(NASH)induced by high-fat diet.Methods:A total of 32 male SPF SD rats was randomly divided into three groups:normal control group,model control group,probiotics intervention group,except for the model control group with 16,the other groups were eight in each.The blank control group was fed with standard feed,and the other two groups were fed with high-fat diet.Two rats from the model control group were killed to evaluate model at random every four weeks.HE staining showed that the rats in the model control group developed nonalcoholic steatosis at the end of fourth weeks,the probiotics group was given the live combined Bifidobacterium and Lactobacillus 0.7 g·kg^(-1) d^(-1) gavage at the beginning of the fifth week.The blank control group and the model control group were given the equal volume of 0.9% Sodium Chloride Solution gastric perfusion,and the total intervention lasted 12 weeks.All the mice were sacrificed 24 hours after last administration,the alanine aminotransferase(ALT),aspartate aminotransferase(AST)contents in serum were detected using biochemical analysis;TG and FFA were tested in homogenate of liver tissue;The plasma endotoxin levels was determined using tachypleus amebocyte lysate(TAL)chromogenic substrate method;The tumor necrosis factor alpha(TNF-α)and Interleukin-1β(IL-1β)levels in serum were detected using ELISA.Results:The rats in the model control group were replicated into NASH models in 16th weeks.Compared with control group,the serum ALT,AST and TG and FFA in the liver homogenate decreased significantly in the probiotic intervention group(P<0.05).The probiotic intervention could improve the level of serum TNF-alpha,IL-1β and plasma LPS(P<0.05)in the serum of NASH model rats.Conclusion:Combined Bifidobacterium and Lactobacillus has some preventive and therapeutic effects on NASH induced by high-fat diet in rats,the mechanism may be related to the reduction of endotoxemia in NASH model rats and the improvement of metabolic inflammation.