摘要
目的基于Keap1/Nrf2/ARE信号通路,研究理中汤对非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)大鼠的保护作用及其机制。方法制备40只NASH大鼠模型,随机分为模型组、理中汤高剂量组、理中汤中剂量组、理中汤低剂量组,每组10只,另设正常组10只。理中汤中剂量组给予理中汤液8.75 g/(kg·d)灌胃,高剂量组予1.5×8.75/(kg·d)灌胃,低剂量予0.5×8.75 g/(kg·d)灌胃;治疗4周。治疗结束后,将各组大鼠处死并观察药物对大鼠血脂、肝功能的影响;肝脏病理学改变情况;ELISA法检测血清炎症细胞因子的表达;Western bolt检测各组大鼠Keap1/Nrf2/ARE信号通路相关蛋白表达;RT-PCR检测相关基因表达。结果模型组大鼠肝脏脂肪变性程度、炎细胞浸润程度最显著,各药物干预组均有不同程度改善,其中以理中汤高剂量组改善最为明显。与正常组比,模型组大鼠肝功能(ALT、AST)血脂(TG、TC)显著升高(P<0.01);各药物干预组均有不同程度改善(P<0.01);其中理中汤高剂量组效果最明显(P<0.01)。模型组大鼠血清IL-6、IL-1β、TNF-α表达较空白组表达显著升高(P<0.01);各药物干预组较模型组明显下降(P<0.01),理中汤高剂量组下降最明显(P<0.01)。模型组大鼠肝组织Keap1、Nrf2、HO-1蛋白表达及基因表达较空白组明显降低,各药物干预组较模型组均有不同程度的上调(P<0.05),其中以理中汤高剂量组蛋白表达及基因表达升高最明显(P<0.01)。结论理中汤能通过上调肝组织中Keap1、Nrf2、HO-1的基因表达,从而改善Keap1/Nrf2/ARE信号通路相关蛋白表达,抑制相关炎症因子的释放,有效改善NASH大鼠肝功能、血脂,达到抗氧化应激的疗效,阻断NASH进展,发挥防治NASH的作用。
Objective Based on the Keap1/Nrf2/ARE signaling pathway,the protective effect of Lizhong Decoction(理中汤)on non-alcoholic steatohepatitis(NASH)in rats and their mechanisms were studied.Methods Forty NASH rats were prepared and randomly divided into model group,Lizhong Decoction high dose group,Lizhong Decoction medium dose group,and Lizhong De⁃coction low dose group(10 rats in each group,and 10 rats in the normal group).Lizhong Decoction high,medium and low dose groups were given the decoction with the corresponding doses[1.5×8.75 g/(kg·d),8.75 g/(kg·d)and 0.5×8.75 g/(kg·d)respectively]by intragastric administration.The treatment lasted for 4 weeks.After the treatment,the rats in each group were kill⁃ed and the effects of drugs on blood lipid and liver function were observed.Pathological changes in the liver were observed.The expression of inflammatory cytokines in serum was detected by ELISA.Western blot method detected the expressions of Keap1/Nrf2/ARE signaling pathway related proteins in each group of rats.RT-PCR was used to detect the expressions of related genes.Results The liver steatosis degree and inflammatory cell infiltration degree of the model group were the most significant,and all the drug intervention groups showed different degrees of improvement,among which the Lizhong Decoction high dose group showed the most obvious improvement.Compared with those of the normal group,liver function(ALT,AST)and blood lipid(TG,TC)of the model group were significantly increased(P<0.01).All drug intervention groups showed different degrees of improvement(P<0.01).The effect was most obvious in Lizhong Decoction groups(P<0.01).The expressions of serum IL-6,IL-1βand TNF-αin the model group were significantly higher than those in the blank group(P<0.01).Compared with those of the model group,those of the drug intervention groups were significantly decreased(P<0.01),and those of the Lizhong Decoction high dose group were decreased most significantly(P<0.01).The protein expressions and gene expressions of Keap1,Nrf2 and HO-1 in rat liver tissues in the model group were significantly lower than those in the blank group,and the protein expressions and gene expressions in each drug intervention group were up-regulated to different levels than those in the model group(P<0.05),among which the protein expressions and gene expressions were increased most significantly in the Lizhong Decoction high dose group(P<0.01).Conclusion Lizhong Decoction can improve the expressions of Keap1,Nrf2 and HO-1 genes in liver tissues,thereby improving the expressions of Keap1/Nrf2/ARE signaling pathway related proteins,inhibiting the release of related inflammatory factors,and ef⁃fectively improving liver function and blood lipid in NASH rats to achieve the effect of anti-oxidative stress,block the progress of NASH,and play the role of prevention and treatment of NASH.
作者
张悦
周晓玲
李泽鹏
陈峭
李灿
韦宛华
刘莹
余静芳
宋征福
ZHANG Yue;ZHOU Xiaoling;LI Zepeng;CHEN Qiao;LI Can;WEI Wanhua;LIU Ying;YU Jingfang;SONG Zhengfu(Liuzhou Hospital of Traditional Chinese Medicine,Liuzhou 545000,Guangxi,China)
出处
《辽宁中医杂志》
CAS
2021年第12期10-14,223,共6页
Liaoning Journal of Traditional Chinese Medicine
基金
国家自然科学基金(81560756)
