NLRP3炎性小体与非酒精性脂肪性肝病的相关性及利拉鲁肽对其的影响

Study of the effect of liraglutide on the correlation between NLRP3 inlammasome and non-alcoholic fatty liver disease|

目的观察核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎性小体与非酒精性脂肪性肝病(NAFLD)的相关性及利拉鲁肽对其的影响。方法选取NAFLD患者(N组)和体检中心健康体检者(C组)各39例,收集其一般资料,测定血清中NLRP3、白细胞介素(IL)-1β,IL-18的水平,分析2组指标差异及其相关性。将30只雄性SD大鼠随机分为正常饮食组(NC组,n=10)和高脂饮食组(HF组,n=20),分别给予普通和高脂饲料喂养:喂养12周后,将HF组随机分为HF组(n=10)和利拉鲁肽组(100L组,n=10),分别给予0.5ml/kg灭菌等渗盐水和100g/kg利拉鲁肽2次/d皮下注射。4周后检测各组血清生化指标和肝脏中NLRP3炎性小体及炎症因子的蛋白表达情况。采用t检验、单因素方差分析(ANOVA)或χ^(2)检验进行统计学分析。结果N组和C组年龄、性别、舒张压、糖化血红蛋白、平均血小板体积、红细胞分布宽度以及血清低密度脂蛋白胆固醇(LDL-Ch)、总胆固醇和总胆汁酸差异无统计学意义。与C组相比,N组的收缩压、体质量指数(BMI)空腹血糖,血肌酐、碱性磷酸酶(ALP).NLRP3、IL-1β、IL-18、甘油三酯、血尿酸、γ-谷氨酰转移酶(GGT),丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和白细胞计数均升高,而高密度脂蛋白胆固醇(HDL-Ch)和总胆红素则低于C组,且差异有统计学意义(P<0.05)。NLRP3与收缩压、BMI、空腹血糖、血肌酐、IL-1β、IL-18、甘油三酯、血尿酸、GGT、ALT、AST呈正相关,但与总胆红素和HDL-Ch呈负相关,差异有统计学意义:与NC组相比,HF组体质量、肝脏质量、血清生化指标(甘油三酯、AST、ALT)、肝脏中NLRP3炎性小体及炎症因子的蛋白表达量均显著升高。经利拉鲁肽治疗后,与HF组相比,100L组各指标均明显下降。结论与健康体检者相比,NAFLD患者炎症相关指标、体质量、血脂和肝功能相关指标均有明显改变,大鼠模型上的研究结果也与此一致;对NAFLD大鼠的利拉鲁肽治疗在一定程度上改善了NFALD,其机制可能是通过调控NLRP3来改善肝脏的炎症及脂肪变性。

Objective To observe the effect of liraglutide on the correlation between nucleotide-binding oligomerization domain-likle receptor family pyrin domain-containing protein 3(NLRP3)inflammasome and nonalcoholic fatty liver disease(NAFLD).Methods Thirty-nine NAFLD cases(group N)and thirty-nine healthy subjects(group C)were selected from the physical examination center,and their general data were colleted to determine the serum levels of NLRP3,IL-1β,and IL-18.The differences and correlations were analyzed between the two sets of indicators.Thirty male SD rats were randomly divided into normal(NC,n=10)and high fat diet group(HF,n=20).The normal group were fed with normal diet and high-fat diet group were fed with high-fat diet.Afier 12 weeks of feeding,HF group was randomly divided into HF group(r=10)and liraglutide group(100L,n=10),and were given 0.5 ml/kg sterile isotonic saline and 100 gkg liraglutide subcutancously twice a day,respectively.Four wecks later,serum biochemical indicators,liver NLRP3 inflammasome protein expression,and inflammatory cytokine conditions were detected in cach group.Statistical analysis was performed using 1 test,oneway analysis of variance(ANOVA)orχ^(2)test.Results There were no statistically significant differenccs between N and c group in tcrms of agc.gender,diastolic blood pressure,glycosylated hemoglobin,mean platelet volumc,crythrocyte distribution width,scrum low-density lipoprotein cholesterol(HDL-Ch).total cholesterol,and tolal bileacid.Compared with group C,group N had clevated systolic blood pressure,body mass index(BMI),fasting blood glucose,blood creatinine,alkaline phosphatase(ALP).NLRP3,interlcukin(IL)-1β,IL-18,TG,blood uric acid,γ-glutamyltransferase(GGT),alanine aminotransferase(ALT),aspartate aminotransferase(AST),and white blood cell counts,while HDL-Ch and total bilirubin were depleted than group C,and the difference was statistically significant(P<0.05).NL RP3 was positively correlated with systolic blood pressure,BMI,fasting blood glucose,serum creatinine,IL-1β,IL-18,triglycerides,serum uric acid,GGT,ALT,AST,but negatively correlated with total bilirubin and HDL-Ch,and the difference was statistically significant.Compared with NC group,HF group had significantly increased body mass,liver mass,serum biochemical indicators(riglycerides,AST,ALT),liver NLRP3 inflammasome protein expression,and inflammatory cytokines.After treatment with liraglutide,100L group indicators were significantly decreased when compared to HF group.Conclusion Compared with healthy subjects,the inflammation-related indicators,body mass,blood lipids and liver fiunction-related indicators are significantly changed in patients with NAFLD,which is also consistent with the results of rat model study.Liraglutide treatment had improved NAFLD to certain extent in NAFLD rats,so NLRP3 regulation may be one of the mechanisms to improve liver inflammation and steatosis.