IL-22在蓝莓益生菌血清干预肝细胞脂肪变性实验中的表达

摘要/Abstract

摘要： 目的探讨蓝莓益生菌血清通过对IL-22表达的影响改善非酒精性脂肪肝的作用机制。方法血清药理学方法制备大鼠10%蓝莓益生菌低、中、高剂量血清。建立游离脂肪酸(FFA)诱导的正常肝细胞株L-02细胞脂肪变性模型,设立正常组、模型组、蓝莓益生菌低、中、高剂量血清组。油红O染色检测细胞内的脂质沉积;定量检测细胞内三酰甘油(TG)含量;RT-PCR检测各组IL-22 基因表达;Western blot及免疫荧光法检测各组IL-22蛋白表达。结果 FFA诱导刺激24 h后,模型组细胞内可见大量脂质沉积,且TG含量明显高于正常组(P=0.000),IL-22的基因及蛋白表达均较正常组明显减弱(P=0.000);与模型组相比,蓝莓益生菌低、中、高剂量血清组的TG逐渐降低(P=0.000),细胞内的脂肪沉积逐渐减轻;高剂量血清组IL-22的基因和蛋白表达较模型组、低剂量血清组、中剂量血清组显著增强(P=0.000),但蓝莓益生菌低、中剂量血清组之间IL-22的基因表达和蛋白表达差异无统计学意义(P>0.05)。结论蓝莓益生菌血清能上调IL-22的表达,参与改善非酒精性脂肪肝的作用机制。

关键词: 蓝莓益生菌血清, 肝细胞, 脂肪变性, 白介素-22

Abstract: Objective To investigate mechanism of blueberry probiotic in treating nonalcoholic fatty liver disease (NAFLD) via affecting expression of interlukin-22 (IL-22).Methods Serum from rats treated with saline, and low, medium and high dose of blueberry probiotic by gavage were collected, respectively. Cell steatosis model was constructed by stimulating normal liver cell line L-02 with free fatty acid (FFA). Serum from different dose blueberry probiotic treated rats were added into cell culture medium as 10% concentration for the L-02 steatosis model, respectively. Intracellular lipid deposition was observed by oil red O staining and level of triglyceride (TG) was quantitatively determined. Gene expression of IL-22 was detected by reverse transcription polymerase chain reaction (RT-PCR) and protein expression of IL-22 was detected by western blot and immunofluorescence.Results After 24-hour FFA stimulation in L-02 cell with serum, a pile of lipid deposition could be observed and TG level was significantly increased (P=0.000), while mRNA and protein levels of IL-22 were decreased (P=0.000). In blueberry probiotic serum groups, TG levels and lipid depositions were negatively correlated with the dose of blueberry probiotic. In high-dose blueberry probiotic serum group, expression of IL-22 was significantly enhanced comparing with the other groups (P=0.000). However, there was no significant difference in expression of IL-22 between the low- and medium-dose serum groups (P>0.05).Conclusion Blueberry probiotic might play a role in treatment of NAFLD by enhancing the expression of IL-22.

Key words: Blueberry probiotic serum, Hepatocyte, Steatosis, Interleukin-22

祝娟娟, 程明亮. IL-22在蓝莓益生菌血清干预肝细胞脂肪变性实验中的表达[J]. 肝脏, 2016, 21(8): 630-635.

ZHU Juan-juan, CHENG Ming-liang. Expression of IL-22 in hepatic steatosis intervened by blueberry probiotic serum[J]. Chinese Hepatolgy, 2016, 21(8): 630-635.

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