摘要
目的 观察不同强度、时间电针对非酒精性脂肪肝病(non-alcoholic fatty liver disease,NAFLD)大鼠肝脏核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)和炎性细胞因子[白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-18(interleukin-18,IL-18)]的影响,探讨其可能的作用机制.方法 将SPF级雄性SD大鼠随机分为对照组和造模组,造模组予高脂饲料喂养5 周后随机分为模型组、假电针组、电针1 组、电针2组,每组15 只,各组再按干预时间的不同(2、3、4 周)分为3个亚组(每组5 只).电针1 组和电针2组均取丰隆和足三里穴进行电针治疗,电流强度分别为 4 mA、2 mA.采用实时荧光定量PCR法检测各组大鼠不同时间点肝脏NLRP3、IL-1β和IL-18 mRNA表达,蛋白质印迹法(Western blot,WB)检测肝脏IL-1β和IL-18蛋白表达,免疫组化法检测肝脏NLRP3 阳性细胞表达.结果 与对照组比较,模型组大鼠肝脏 NLRP3、IL-1β和 IL-18 mRNA 与蛋白表达均显著升高(P＜0.01).与模型组、假电针组比较,电针1 组和电针2组干预2、3、4周后肝脏NLRP3、IL-1β和IL-18 mRNA与蛋白表达均降低(P＜0.05).与电针 1 组比较,电针 2 组 2、3、4 周后大鼠肝脏NLRP3、IL-1β和IL-18 mRNA与蛋白表达均升高(P＜0.05).同组内各时间点比较,电针 1 组干预 4 周后肝脏NLRP3、IL-1β和IL-18 mRNA与蛋白表达均最低(P＜0.05).结论 不同强度、时间电针可影响NAFLD大鼠治疗效果,其中电流强度 4 mA且干预4 周的作用更佳,其机制可能与抑制NLRP3 炎性小体激活、减少炎性细胞因子、抑制炎症反应有关.
Abstract
Objective To observe the effects of electroacupuncture(EA)of different intensities and durations on nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)and inflammatory cytokines interleukin-1β(IL-1β)and interleukin-18(IL-18)in the liver of rats with non-alcoholic fatty liver disease(NAFLD)and discuss the potential mechanism.Method Male Sprague-Dawley(SD)rats of SPF degree were randomized into a control group and a modeling group.After 5-week high-fat feeding,rats in the modeling group were randomly allocated into a model group,a sham EA group,EA group 1,and EA group 2,with 15 rats in each group;each group was further divided into 3 sub-groups(5 rats in each group)based on the intervention time(2,3,and 4 weeks).Bilateral Fenglong(ST40)and Zusanli(ST36)were selected for EA intervention in EA group 1 and EA group 2,and the current intensity was 4 mA and 2 mA,respectively.At each time point,quantitative real-time polymerase chain reaction(PCR)was used to detect the mRNA expression of NLRP3,IL-1β,and IL-18 in the liver of each group of rats,Western blot(WB)was adopted to determine the protein expression of IL-1β and IL-18 in the liver,and immunohistochemistry was used to detect the expression of NLRP3 positive cells in the liver.Result Compared to the control group,the mRNA and protein expression levels of NLRP3,IL-1β,and IL-18 increased significantly in the liver of rats in the model group(P＜0.01).Compared to the model and sham EA groups,the mRNA and protein expression levels of liver NLRP3,IL-1β,and IL-18 dropped in EA group 1 and EA group 2 after intervention weeks 2,3,and 4,respectively(P＜0.05).Compared to EA group 1,the mRNA and protein expression levels of liver NLRP3,IL-1β,and IL-18 increased in EA group 2 after intervention weeks 2,3,and 4,respectively(P＜0.05).Intra-group comparisons among different time points showed that the lowest mRNA and protein expression levels of liver NLRP3,IL-1β,and IL-18 were found after intervention week 4 in EA group 1(P＜0.05).Conclusion Different intensities and durations can affect the efficacy of EA in intervening NAFLD rats,and EA with a current intensity of 4 mA for 4 weeks should be optimal;the mechanism may be associated with inhibiting the activation of NLRP3 inflammasome,reducing inflammatory cytokines,and suppressing inflammatory responses.
维普
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