基于NF-κB/NLRP3/IL-1β信号通路探讨石斛合剂对糖尿病伴非酒精性脂肪性肝病大鼠细胞炎症反应及凋亡的作用  

Effect of Dendrobii Caulis Mixture on Cell Inflammatory Response and Apoptosis in Diabetic Rats with Nonalcoholic Fatty Liver Disease Based on NF-κB/NLRP3/IL-1βSignaling Pathway

作　　者：邹玉卿 施红[1,2] 刘欣 庄舒婷[1] 张家林 陈佳敏 张捷平 ZOU Yuqing;SHI Hong;LIU Xin;ZHUANG Shuting;ZHANG Jialin;CHEN Jiamin;ZHANG Jieping(Fujian University of Traditional Chinese Medicine(TCM),Fuzhou 350000,China;The Third People's Hospital Affiliated to Fujian University of TCM,Fuzhou 350000,China)

机构地区：[1]福建中医药大学,福州350000 [2]福建中医药大学附属第三人民医院,福州350000

出　　处：《中国实验方剂学杂志》2023年第18期78-86,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81973827)。

摘　　要：目的:探讨石斛合剂对糖尿病伴非酒精性脂肪肝模型大鼠细胞炎症性反应及凋亡的影响及可能作用机制。方法:雄性SD大鼠40只,按体质量随机分为空白组10只和2型糖尿病(T2DM)伴非酒精性脂肪性肝病(NAFLD)模型组30只。采用高糖高脂饮食联合腹腔注射链脲佐菌素(STZ)建立模型,按血糖、体质量随机分为模型组,石斛合剂组(16.67 g·kg^(-1)·d^(-1)),二甲双胍组(100 mg·kg^(-1)·d^(-1)),每组10只,按剂量连续灌胃给药4周,空白组与模型组灌胃按10 mL·kg^(-1)·d^(-1)给予生理盐水。检测各组大鼠空腹血糖(FBG)、血清胰岛素(INS)、糖化血清蛋白(GSP)、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)等指标。各组大鼠肝组织采用苏木素-伊红(HE)染色观察病理改变,免疫组织化学法观察核转录因子(NF)-κB、NOD样受体热蛋白结构域相关蛋白3(NLRP3)、白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α阳性表达。蛋白免疫印迹法(Western blot)、实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠肝组织B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶-3(Caspase-3)、NF-κB p65(NF-κB p65)、NLRP3、IL-1β、TNF-α的蛋白及mRNA表达情况。结果:与空白组比较,模型组FBG、GSP、TC、TG、LDL-C、AST、ALT均上升,INS、HDL-C下降,Bax、Caspase-3、NLRP3、IL-1β、TNF-α蛋白及mRNA表达上升,p-NF-κB/NF-κB蛋白上升,Bcl-2蛋白及mRNA表达下降,NF-κB、NLRP3、IL-1β、TNF-α免疫组化阳性表达升高,差异有统计学意义(P<0.01),肝脏形态结构被破坏,可见明显脂肪空泡。与模型组比较,石斛合剂组与二甲双胍组FBG、GSP、TC、TG、LDL-C、AST、ALT均下降,INS、HDL-C上升,Bax、Caspase-3、NLRP3、IL-1β、TNF-α蛋白及mRNA表达下降,p-NF-κB/NF-κB蛋白下降,Bcl-2蛋白及mRNA表达升高,NF-κB、NLRP3、IL-1β、Objective:To investigate the effect of Dendrobii Caulis mixture on cell inflammatory response and apoptosis in diabetic rat with non-alcoholic fatty liver disease(NAFLD)and its possible mechanism.Method:Forty male SD rats were randomly divided into blank group and model group of type 2 diabetes mellitus(T2DM)with NAFLD according to body weight.The model was established by high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin(STZ),which was randomly divided into the model group,Dendrobii Caulis mixture group(16.67 g·kg^(-1)·d^(-1))and metformin group(100 mg·kg^(-1)·d^(-1))according to blood glucose and body weight,and 10 rats in each group.Rats in each group were administered by continuous gavage for 4 weeks,the blank and model groups were given saline by gavage at 10 mL·kg^(-1)·d^(-1).Fasting blood glucose(FBG),serum insulin(INS),glycosylated serum protein(GSP),triglyceride(TG),total cholestero(l TC),high density lipoprotein(HDL-C),low density lipoprotein(LDL-C),alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were detected in each group of rats.The liver tissues of rats in each group were stained with hematoxylin-eosin(HE)to observe the pathological changes,and the positive expressions of nuclear transcription factor(NF)-κB,NOD-like receptor heat protein structural domain-related protein 3(NLRP3),interleukin(IL)-1βand tumor necrosis factor(TNF)-αwere observed by immunohistochemistry.Western blot and fluorescence quantitative polymerase chain reaction(Real-time PCR)were used to detect the protein and mRNA expression of B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),Caspase-3 and NF-κB p65,NLRP3,IL-1β,TNF-αin liver tissue of rats in each group.Result:Compared with the blank group,FBG,GSP,TC,TG,LDL-C,AST and ALT levels were increased,INS and HDL-C levels were decreased,Bax,Caspase-3,NLRP3,IL-1β,TNF-αprotein and mRNA expression were increased in model group,the ratio of p-NF-κB/NF-κB protein increased,the expression of Bcl-2 protein and mRNA d

关 键 词：石斛合剂 2型糖尿病(T2DM) 非酒精性脂肪性肝病(NAFLD) 炎症反应 细胞凋亡 核转录因子-κB(NF-κB) 白细胞介素 

分 类 号：R22[医药卫生—中医基础理论] R587.1[医药卫生—中医学] R28R96