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目的探讨清化方对非酒精性脂肪肝(NAFLD)大鼠辅助性T细胞17(Th17)/调节性T细胞(Treg)平衡及相关细胞因子的影响。方法将60只雄性Wistar大鼠分为空白组、黄连素对照组及低、中、高清化方组,每组10只。空白组给予普通饲料,模型组及中药组喂养高脂饲料。治疗组予清化方药液灌胃,模型组予黄连素液灌胃,空白组灌服等量的0.9%NaCl溶液,各组均干预12周。实验结束后,采用流式细胞术检测大鼠外周血及肠道组织Th17/Treg细胞比例,并且测定大鼠血清肝功能(ALT、AST、GGT)、血脂(TG、TC、HDL、LDL),血浆内毒素(LPS)及相关细胞因子(TGF-β、TNF-α、IL-6、IL-8、IL-10、IL-17)水平;采集大鼠肝组织,HE染色后观察肝脂肪变性情况。结果 NAFLD模型大鼠外周血Th17细胞比率及相关细胞因子(IL-17、IL-6、IL-8)较空白组明显增高(P<0.05),清化方中高剂量组、中剂量组及对照组较模型组显著降低(P<0.05)。模型组NAFLD大鼠肠道固有层Treg细胞比例显著低于空白组(P<0.05);模型组大鼠肠道固有层Th17/Treg细胞比例高于空白组(P<0.05)。结论NAFLD大鼠模型外周血中Th17/Treg比例及相关的细胞因子水平高于空白组,清化方能够通过调节Th17/Treg失衡而减轻NAFLD大鼠肝细胞的炎症和肝脏的脂肪沉积。
Objective To investigate the effects of Qinghua Recipe on the balance of Th17 / Tregs and related cytokines in non-alcoholic fatty liver disease (NAFLD) rats. Methods Sixty male Wistar rats were divided into blank group, berberine control group and low, middle and high-grade group, 10 rats in each group. The blank group was given normal feed, and the model group and the traditional Chinese medicine group were fed with high fat diet. The treatment group was given qinghuafang liquid, and the model group was treated with berberine solution. The blank group was fed with the same amount of 0.9% NaCl solution, and each group was treated for 12 weeks. After the experiment, the proportion of Th17 / Treg cells in peripheral blood and intestinal tissue of rats was determined by flow cytometry. The serum levels of ALT, AST, GGT, TG, TC, HDL, LDL, The levels of plasma LPS and related cytokines (TGF-β, TNF-α, IL-6, IL-8, IL-10 and IL-17) Happening. Results Compared with the blank group, the Th17 cell ratio and related cytokines (IL-17, IL-6, IL-8) in peripheral blood of NAFLD model group were significantly increased Group than the model group was significantly lower (P <0.05). The proportion of Treg cells in intestinal lamina propria of NAFLD model group was significantly lower than that in blank group (P <0.05). The proportion of Th17 / Treg cells in intestinal lamina propria of model group was higher than that of blank group (P <0.05). Conclusions The proportion of Th17 / Treg and related cytokines in peripheral blood of NAFLD rats is higher than that of blank control group. Qinghua Prescription can reduce the inflammation of hepatic cells and the fat deposition of liver in NAFLD rats by regulating the imbalance of Th17 / Treg.