茱萸丸及其拆方对动脉粥样硬化模型小鼠主动脉斑块及主动脉组织NLRP3/ASC/Caspase-1信号通路的影响  

Effects of Zhuyu Pill(茱萸丸)and its Ingredient on Aortic Plaque and the NLRP3/Asc/Caspase-1 Signaling Pathway in Aortic Tissue of Mice in Atherosclerosis Model

作　　者：宋玮 张钟艺 王楷 沈涛[1] SONG Wei;ZHANG Zhongyi;WANG Kai;SHEN Tao(Chengdu University of Traditional Chinese Medicine,Chengdu,610075)

机构地区：[1]成都中医药大学,四川省成都市610075

出　　处：《中医杂志》2024年第6期618-626,共9页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81973743);四川省科技计划项目(2022YFS0381,2020JDRC0103)。

摘　　要：目的探讨茱萸丸治疗动脉粥样硬化的可能作用机制及配伍意义。方法13只正常C57BL/6J小鼠作为空白组,40只同品系的ApoE-/-小鼠随机分为模型组、黄连组、吴茱萸组、茱萸丸组各10只。除空白组外,其余各组均采用高脂饲料连续12周喂养制备动脉粥样硬化模型。造模后黄连组给予黄连药液143.34 mg/(kg·d)灌胃,吴茱萸组给予吴茱萸药液301.78 mg/(kg·d)灌胃,茱萸丸组给予茱萸丸药液261.07 mg/(kg·d)灌胃,空白组及模型组每日给予0.3 ml生理盐水灌胃,各组小鼠均灌胃12周。各组小鼠采用生化法检测总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、空腹血糖水平;采用ELISA检测血清中空腹胰岛素,并计算胰岛素抵抗指数;采用HE染色观察小鼠主动脉组织、肝脏、附睾脂肪的病理学形态变化,油红O染色观察小鼠主动脉组织脂质沉积情况,并计算主动脉斑块面积占比、主动脉红染脂质面积占比、非酒精性脂肪肝活动性积分(NAS)评分、附睾脂肪横截面面积;采用ELISA检测血清中白细胞介素1β(IL-1β)、白细胞介素18(IL-18)水平;采用Western blot检测主动脉组织核苷酸结合寡聚化结构域样受体家族吡啉结构域蛋白3(NLRP3)蛋白表达;采用免疫荧光法检测含CARD结构域的凋亡相关颗粒样蛋白(ASC)、含半胱氨酸的天冬氨酸蛋白酶1(Caspase-1)蛋白表达;采用实时荧光PCR法检测小鼠主动脉组织NLRP3、ASC、Caspase-1、IL-1β、IL-18 mRNA表达量。结果与空白组比较,模型组血清TC、TG、LDL-C升高,空腹胰岛素降低,空腹血糖、胰岛素抵抗指数升高(P<0.01);主动脉斑块增大,肝脏脂质沉积增多,附睾脂肪细胞体积变大,主动脉斑块面积占比及红染脂质面积占比增加,肝脏NAS评分升高,附睾脂肪横截面面积变大,血清IL-1β、IL-18升高,主动脉组织NLRP3、ASC、Caspase-1蛋白及mRNA表达水平升高(P<0.01)。与模型组比较,黄连组、吴茱萸组、Objective To investigate the possible mechanism of action and combination of medicinals of Zhuyu Pill(茱萸丸)in the treatment of atherosclerosis.Methods Thirteen normal C57BL/6J mice were used as blank group,and 40 ApoE-/-mice of the same strain were randomly divided into model group,Huanglian(Coptis chinensis Franch.)group,Wuzhuyu(Tetradium ruticarpum)group,and Zhuyu Pill group,with 10 mice in each group.Except the blank group,the other groups were fed with high-fat diet for modeling,and the total modeling cycle was 12 weeks.After modelling,Huanglian group was given Huanglian solution 143.34 mg(/kg·d),Wuzhuyu group with Wuzhuyu solution 301.78 mg(/kg·d),Zhuyu Pill group with 261.07 mg(/kg·d)of Zhuyu Pill solution,blank group and model group were given 0.3ml of normal saline.Mice in all groups were gavaged for 12 weeks.The levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C)and fasting blood glucose were measured by biochemical method;fasting insulin in serum was detected by enzyme-linked immunosorbent assay(ELISA)and insu⁃lin resistance index was calculated;HE staining was used to observe the pathological and morphological changes of aortic tissue,liver and epididymal fat in mice,and oil red O staining was used to observe the lipid deposition of aortic tissue in mice,and the percentage of positive area of aortic HE staining,the percentage of positive area of aortic oil red O staining,the non-alcoholic fatty liver activity score(NAS)score,and the cross-sectional area of epididymal fat were calculated;serum levels of interleukin 1β(IL-1β)and interleukin 18(IL-18)were detected by ELISA;pro⁃tein expression of nucleotide-binding oligomerisation structural domain-like receptor family pyridoxal structural domain protein 3(NLRP3)in aortic tissues was detected by Western blot;and immunofluorescence was used to detect the levels of apoptosis-associated granulocyte-like protein(ASC),which contains the CARD structural domain(ASC)and cysteine-containing aspartic protease 1(C

关 键 词：动脉粥样硬化 主动脉斑块 茱萸丸 核苷酸结合寡聚化结构域样受体家族吡啉结构域蛋白3 含CARD结构域的凋亡相关颗粒样蛋白 含半胱氨酸的天冬氨酸蛋白酶1 

分 类 号：R285.5[医药卫生—中药学]