定心方Ⅲ号方调控JAK2/STAT3通路对肥胖小鼠脂肪炎症的影响  

Dingxin RecipeⅢAttenuates Inflammation of Adipose Tissue in Obese Mice by Regulating JAK2/STAT3 Pathway

作　　者：张丽芳 胡芳 程赛博[1] 刘晓瑜 何培坤 朱严欣 李钊泳 贾钰华[1] ZHANG Li-fang;HU Fang;CHENG Sai-bo;LIU Xiao-yu;HE Pei-kun;ZHU Yan-xin;LI Zhao-yong;JIA Yu-hua(School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515;Department of Traditional Chinese Medicine,Pingshan General Hospital of Southern Medical University,Guangdong 518118)

机构地区：[1]南方医科大学中医药学院,广州510515 [2]南方医科大学坪山总医院中医科,广东518118

出　　处：《中国中西医结合杂志》2022年第8期979-986,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81973802,No.82174299);广东省自然科学基金资助项目(No.2021A1515011603)。

摘　　要：目的探讨定心方Ⅲ号方(DXRⅢ)调控蛋白质酪氨酸激酶2(JAK2)/信号转导和转录激活因子3(STAT3)信号通路对肥胖小鼠脂肪组织慢性炎症的影响。方法32只8周龄雄性ApoE-/-小鼠按随机数字表法分为模型组,DXRⅢ低、中、高剂量组,每组8只,另取8只同龄C57BL/6J雄性小鼠作为正常组。ApoE-/-小鼠连续喂食西方饮食饲料12周建立肥胖模型后,DXRⅢ低、中、高剂量组分别予6.5、13、26 g/kg DXRⅢ中药煎剂灌胃,正常组和模型组以等量生理盐水灌胃。各组均每天给药1次,连续给药12周。每周测量小鼠体重。干预结束后,取小鼠内脏脂肪组织称重并计算内脏脂肪指数;采用生化法检测小鼠血清TC、TG、LDL-C、HDL-C水平;HE染色观察小鼠内脏脂肪组织病理学变化;IL-1β免疫组化染色分析内脏脂肪组织炎症浸润程度;RT-PCR检测脂肪组织炎症因子IL-1β、IL-6、肿瘤坏死因子-α(TNF-α)、单核细胞趋化蛋白-1(MCP-1)mRNA表达,巨噬细胞极化标志物诱导型一氧化氮合酶(iNOS)和精氨酸酶-1(Arg-1)mRNA表达;Western Blot检测小鼠脂肪组织JAK2、磷酸化JAK2(p-JAK2)、STAT3、磷酸化STAT3(p-STAT3)、iNOS、Arg-1蛋白表达。结果与正常组比较,模型组小鼠体重、体重增加值、内脏脂肪重量、内脏脂肪指数与血清中TC、TG、LDL-C、HDL-C水平均升高(P<0.05,P<0.01),HE染色显示脂肪细胞肥大,免疫组化染色显示IL-1β水平上调,脂肪组织中IL-1β、IL-6、TNF-α、MCP-1、iNOS mRNA表达增加,iNOS蛋白、JAK2、STAT3蛋白磷酸化水平亦增加(P<0.05,P<0.01),Arg-1 mRNA及蛋白表达降低(P<0.05);与模型组比较,DXRⅢ各剂量组小鼠体重增加值、内脏脂肪重量、内脏脂肪指数与血清中TC、LDL-C水平均下降(P<0.05,P<0.01),HE染色显示脂肪细胞变小,免疫组化染色显示IL-1β水平下调;DXRⅢ低、中剂量组小鼠血清TG水平下降(P<0.05),脂肪组织IL-1β、IL-6、iNOS mRNA表达下降,iNOS蛋白、JAK2、STAT3蛋白磷酸化�Objective To explore the effect of Dingxin RecipeⅢ(DXRⅢ)on chronic inflammation of adipose tissue in obese mice by regulating the Janus protein tyrosine protein kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway.Methods Thirty-two 8-week-old male ApoE-/-mice were divided into the model group,low,medium,high dose DXRⅢgroups by random digit table,with 8 in each group,and another 8 C57BL/6J male mice of the same age were recruited as normal group.ApoE-/-mice were fed with Western diet for 12 weeks to establish the obesity model.Mice in the low,medium,high dose DXRⅢgroups were administered with 6.5,13 and 26 g·kg-1 DXRⅢrespectively and equal volume of distilled water were administered to mice in the model and normal group.All the above groups were administered once a day for consecutive 12 weeks.Monitor the body weight of the mice weekly.After the intervention,all visceral adipose tissue were carefully collected and calculated the visceral adipose tissue index.Serum TC,TG,LDL-C and HDL-C levels were measured with the method of biochemical kits.HE staining was used to observe the histopathology of the visceral adipose tissue.IL-1βimmunohistochemistry was performed to evaluate the degree of inflammatory infiltration.The IL-1β,IL-6,TNF-α,monocyte chemoattractant protein-1(MCP-1),inducible nitric oxide synthase(iNOS)and arginase-1(Arg-1)mRNA expression were detected by RT-PCR.Protein levels of iNOS,Arg-1,JAK2,p-JAK2,STAT3 and p-STAT3 were assessed by Western Blot.Results Compared with the normal group,the body weight,weight gain,the weight of viceral adipose tissue,visceral adipose tissue index,and the serum levels of TC,TG,LDL-C and HDL-C increased(P<0.05,P<0.01),HE staining showed adipocyte hypertrophy,immunohistochemical staining showed the level of IL-1βincreased,the mRNA expressions of IL-1β,IL-6,TNF-α,MCP-1,iNOS increased,protein level of iNOS and the phosphorylation of JAK2 and STAT3 also increased(P<0.05,P<0.01),while Arg-1 mRNA and protein decreased(P<0.05)

关 键 词：定心方Ⅲ号方 脂肪组织炎症 巨噬细胞极化 蛋白质酪氨酸激酶2 信号转导和转录激活因子3 

分 类 号：R285.5[医药卫生—中药学]