摘要
目的探讨紫薯花青素通过核因子κB(NF-κB)通路减轻高脂饲料联合四氯化碳诱导大鼠脂肪性肝炎机制。方法将70只雄性大鼠随机分为对照组(10只)和高脂饲料组(60只),对照组喂饲普通饲料,高脂组喂饲高脂饲料同时每周2次腹腔注射四氯化碳-橄榄油制剂(50:50)2 mL/kg,对照组注射等量橄榄油,共10周,每周称体重并检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、甘油三酯(TG)和总胆固醇(TC)浓度。从造模成功的58只大鼠中随机选取50只再分为模型组,紫薯花青素低、中、高剂量组(60、120、240 mg/kg)和阳性药物组(水飞蓟素150 mg/kg),每组10只,每天1次等体积(20 mL)灌胃给药,对照组和模型组灌胃等量生理盐水,8周后,检测大鼠血清ALT、AST、TC、TG、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)浓度和血清促炎症因子肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和抗炎症因子白细胞介素4(IL-4)、白细胞介素13(IL-13)浓度;实时定量荧光聚合酶链式反应(PCR)检测大鼠肝脏内TNF-α、IL-1β、IL-4、IL-13、过氧化物酶体增殖物激活受体(PPAR-γ)和高迁移率族蛋白B1(HMGB-1)的mRNA表达;Western blotting检测肝组织核因子κB抑制蛋白(IκB)磷酸化程度及PPAR-γ和HMGB-1蛋白表达。结果与对照组比较,造模后大鼠体重增加(P<0.05),血清AST、ALT、TG、TC和LDL浓度均显著升高(P<0.05),HDL浓度明显下降(P<0.05);与对照组相比,模型组大鼠TNF-α、IL-1β和HMGB-1的表达显著升高(P<0.05),IL-4、IL-13和PPAR-γ的表达显著降低(P<0.05),磷酸化IκB(p-IκB)和NF-κB表达量明显增高(P<0.05);与模型组比较,高剂量紫薯花青素组NF-κB的表达水平明显降低,IκB磷酸化程度下降(P<0.05),肝组织内TNF-α、IL-1β和HMGB-1 mRNA表达量显著降低(P<0.05),IL-4、IL-13和PPAR-γ的mRNA表达显著升高(P<0.05)。紫薯花青素低、中、高剂量组及阳性药物组上述指标均有所改善,高剂量组作用优于低、中剂量组,与阳性药物组效果接近。结论紫薯花青素可通过NF-κB通路起到减轻高脂饲料联合四氯化碳诱导大鼠脂肪性肝炎的作用。
Objective To explore the mechanism of purple sweet potato anthocyanins through NF-κB pathway in attenuating steatohepatitis induced by high fat diet combined with carbon tetrachloride in rats.Methods Seventy male rats were randomly divided into control group（ n = 10） and high-fat diet group（ n = 60）,models were prepared by highfat diet and intraperitoneal injection of carbon tetrachloride and olive oil（ 50 ： 50） 2 mL/kg,two times a week.After 10 weeks of feeding,the weight variations of all rats were tested before and after modeling.The colorimetric technique was used to test the concentration of serum ALT,AST,TG,and TC.A total of 58 rats were succeeded in modeling,the random choice of 50 rats were divided into model group,purple sweet potato anthocyanin low dose group（ 60 mg/kg）,middle dose group（ 120 mg/kg）,high dose group（ 240 mg/kg）,positive drug group（ 150 mg/kg）,10 rats in each group.After 8 weeks of continuous administration,the method of colorimetric technique was used to test the concentration of ALT,AST,TG,TC,HDL and LDL.The method of ELISA kit was used to test the levels of the pro-inflammatory factors TNF-α,IL-1β and the levels of the anti-inflammatory IL-4,IL-13.The Real-time PCR was used to test the expression of TNF-α,IL-1β,IL-4,IL-13,PPAR-γ and HMGB-1 mRNA of rats liver.The western blotting method was adopted to test the level of IκB phosphorylation in liver tissues as well as the PPAR-γ and HMGB-1 protein expression.Results After modeling,the body weight of rats increased（ P 〈 0.05）,the concentration of serum insulin AST,ALT,TG,TC,and LDL all increased significantly（ P 〈 0.05）,the concentration of HDL decreased obviously（ P 〈 0.05）.Compared with control group,the expression of TNF-α,IL-1β and HMGB-1 in model group were increased significantly（ P 〈 0.05）,the expression of IL-4,IL-13 and PPAR-γ were decreased significantly（ P 〈 0.05）,the expression of pIκB and NF-κB in the model group increased significantly（ P 〈 0.05）.Compared with the model group,the expression level of NF-κB in the high dose purple sweet potato anthocyanin group decreased obviously,and the phosphorylation degree of IκB decreased（ P 〈 0.05）,the mRNA expression of TNF-α,IL-1β and HMGB-1 in liver tissue was significantly decreased（ P 〈 0.05）,and the mRNA expression of IL-4,IL-13 and PPAR-γ were significantly increased（ P 〈 0.05）.The purple sweet potato anthocyanin low dose group,middle dose group,high dose group and positive drugs group all improved the above indexes in different degrees.The effect of the high dose group was significantly higher than that of the low dose and medium dose group,but equivalent to that of the positive drug group.Conclusion Purple sweet potato anthocyanins through NF-κB pathway have a role in attenuating steatohepatitis induced by high fat diet combined with carbon tetrachloride in rats.
作者
宓伟
韩富磊
梁洁
梁亚楠
关百初
徐昊
Mi Wei;Han Fulei;Liang Jie;Liang Yanan;Guan Baichu;Xu Hao(Nutrition and Food Hygiene Section,Public Health and Management College,Binzhou Medical University,Yantai 264003,Chin)
出处
《卫生研究》
CAS
CSCD
北大核心
2018年第4期517-524,共8页
Journal of Hygiene Research
基金
山东省高校科技计划项目(No.J17KA224)
