凯里酸汤对代谢性脂肪肝的改善作用及其机制探讨

Improvement of Kaili Sour soup on metabolic fatty liver and its mechanism

目的:探讨凯里酸汤对代谢性脂肪肝(MAFLD)的改善作用及其机制。方法:34只SD大鼠随机分为4组,正常对照(ND,n=8)组、正常饮食+酸汤(NS,n=8)组、模型对照(HF,n=10)组和高脂饮食+酸汤(HS,n=8)组。ND和NS组大鼠给予标准饲料喂养,HF和HS组大鼠给予高脂饲料喂养以诱导MAFLD模型。造模同时,NS组和HS组大鼠给予酸汤灌胃干预,ND组和HF组大鼠给予等体积双蒸水灌胃。灌胃酸汤当日煮熟,自然冷却后灌胃,灌胃剂量1 ml/100 g,6 d/周,1 d/次。饲喂12周后,处死各组大鼠,收取血和肝组织样本。称量各组大鼠体质量及肝质量,计算肝指数和Lee’s指数;测定血清AST、ALT活性、TG、TC、HDL-C、LDL-C和TBA含量变化;HE和油红O染色观察肝组织病理形态变化及脂肪变性情况;粪便16SrRNA测序检测大鼠肠道菌群变化;Western blot和IHC检测IL-6和TNF-α表达变化;Western blot检测TLR-4和NF-κB p65表达变化。结果:与ND组比较,HF组大鼠体质量、肝质量、肝指数和Lee’s指数显著升高(P<0.05);血清中HDL-C含量显著降低,其余血清生化指标活性/含量均显著升高(P<0.05);肝小叶界限不清晰,肝组织发生明显病理性改变及脂肪变性;大鼠肠道菌群及菌群结构均发生了显著改变,粪便肠道菌群的丰度和多样性均显著增加;肝组织中炎症因子(IL-6、TNF-α)蛋白表达明显增加(P<0.05);TLR-4和NF-κB p65蛋白表达明显增加(P<0.05)。与HF组比较,HS组大鼠上述指标均明显改善(P<0.05),肝组织中脂滴明显减少或变小。结论:凯里酸汤能有效改善肠道微生态环境,进而改善胆汁酸及脂质代谢,对MAFLD具有较好的改善作用。

Objective:To explore the improvement effect of Kaili Sour soup on metabolic fatty liver(MAFLD)and its mechanism.Methods:Thirty-four SD rats were randomly divided into four groups:normal control group(ND,n=8),normal diet plus sour soup group(NS,n=8),model control group(HF,n=10)and high-fat diet plus sour soup group(HS,n=8). The rats in ND and NS groups were fed with standard diet,and the rats in HF and HS groups were fed with high fat diet to induce MAFLD model. At the same time,rats in NS group and HS group were given sour soup and rats in ND group and HF group were given the same volume of double distilled water. The gastric acid decoction was cooked on the same day,cooled naturally,and then given to the stomach at a dose of1 ml/100 g,6 d/week,1 d/time. After feeding for 12 weeks,rats in each group were killed and blood and liver tissue samples were collected. Weighed the body mass and liver mass of rats in each group and calculated the liver index and Lee’s index;serum AST,ALT activity,TG,TC,HDL-C,LDL-C and TAB were measured. HE and Oil Red O staining were used to observe the pathological changes and steatosis of liver tissue. Intestinal flora changes in rats was detected by fecal 16SrRNA sequencing.The expression changes of inflammatory factors(IL-6 and TNF-α)were detected by Western blot and IHC. The expression changes of TLR-4 and NF-κB p65 were detected by Western blot.Results:Compared with ND group,the body mass,liver mass,liver index and Lee’s index of HF group were significantly increased(P<0.05). The content of HDL-C in serum decreased significantly,and the activity/content of other serum biochemical indexes increased significantly(P<0.05). The boundaries of liver lobules were unclear,and obvious pathological changes and steatosis occured in liver tissue;the intestinal flora and structure of rats changed significantly,and the abundance and diversity of fecal intestinal flora increased significantly;the expressions of inflammatory markers(IL-6,TNF-α)in liver tissue increased significantly(P<0.05). The expressions of TLR-4 and NF-κB p65 in liver tissue increased significantly(P<0.05). Compared with HF group,the above indexes in HS group were significantly improved(P<0.05). Lipid droplets in liver tissue decreased or became smaller obviously.Conclusion:Kaili Sour soup can effectively improve intestinal microecological environment,and then improve bile acid and lipid metabolism,and finally alleviate MAFLD.