胆瘀方药液对大鼠主动脉内皮细胞脂质泡沫化的抑制作用及其机制  

Inhibitory effect and mechanism of Danyu formula on oxidized low-density lipoprotein-induced foam cell formation in rat aortic endothelial cells

作　　者：宋晓龙[1] 潘仁友 涂冬云 顾月星[1] 宋峻 伍德明[1] 倪其猛[1] SONG Xiaolong;PAN Renyou;TU Dongyun;GU Yuexing;SONG Jun;WU Deming;NI Qimeng(Department of Cardiology,Yancheng Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine,Yancheng 224001,China)

机构地区：[1]南京中医药大学附属盐城中医院盐城市中医院心内科,江苏盐城224001

出　　处：《山东医药》2023年第35期39-44,共6页Shandong Medical Journal

基　　金：江苏省老年健康科研项目(LKM2022076)。

摘　　要：目的 探讨胆瘀方药液对氧化低密度脂蛋白(Ox-LDL)诱导大鼠主动脉内皮细胞(RAECs)脂质泡沫化的抑制作用及其可能的机制。方法 将RAECs随机分为对照组、模型组、胆瘀方低剂量组、胆瘀方中剂量组、胆瘀方高剂量组及阿托伐他汀组,除对照组外,其余五组均灌胃给予200μg/mL的Ox-LDL作用72 h,胆瘀方低、中、高剂量组及阿托伐他汀组分别灌胃给予50、100、200μg/L胆瘀方药液及阿托伐他汀300μg/L。各组分组处理后24、48、72 h,采用CCK-8法检测细胞增殖能力;取分组处理72 h的各组细胞,油红O染色观察细胞脂质泡沫化情况,ELISA法检测细胞肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)蛋白及细胞上清液丙二醛(MDA)、超氧化物歧化酶(SOD)表达,二羟甲基荧光素氧化荧光反应检测细胞活性氧(ROS)含量,实时荧光定量PCR法检测细胞蛋白质酪氨酸激酶2(JAK2)、信号转导与转录激活因子3(STAT3)mRNA相对表达量,Westen blotting法检测细胞JAK2、p-JAK2、STAT3、p-STAT3蛋白相对表达量,并计算p-JAK2/JAK2、p-STAT3/STAT3。结果 与对照组作用相同时间点比较,模型组OD值均降低(P均<0.05);与模型组作用相同时间点比较,胆瘀方高剂量组及阿托伐他汀组OD值均升高,胆瘀方中剂量组作用48、72 h的OD值均升高(P均<0.05)。对照组细胞无脂质泡沫化情况,模型组细胞脂质泡沫化明显;与模型组比较,阿托伐他汀组及胆瘀方高、中、低剂量组细胞脂质泡沫化均得到改善,胆瘀方高剂量组和阿托伐他汀组细胞脂质泡沫化缓解最明显,几乎趋近于对照组。与对照组比较,模型组细胞TNF-α、IL-6、ROS、JAK2 mRNA相对表达量、STAT3 mRNA相对表达量、p-JAK2/JAK2、p-STAT3/STAT3及上清液MDA表达均升高,上清液SOD表达降低(P均<0.05)。与模型组比较,阿托伐他汀组及胆瘀方高、中、低剂量组细胞TNF-α、IL-6、ROS、JAK2 mRNA相对表达量、STAT3 mRNA相对表达量、p-JAK2/JAK2Objective To explore the inhibitory effect of Danyu formula on foam cell formation in rat aortic endothelial cells(RAECs)induced by oxidized low-density lipoprotein(Ox-LDL)and its possible mechanism.Methods RAECs were randomly divided into the control group,model group,low-dose Danyu formula group,medium-dose Danyu formula group,and high-dose Danyu formula group,and atorvastatin group.Except for the control group,RAECs in the rest of the five groups were treated with 200μg/mL Ox-LDL for 72 h,and the low-,medium-,and high-dose Danyu formula groups and the atorvastatin group were treated with 50,100,and 200μg/L Danyu formula solution and 300μg/L atorvastatin,respectively.After 24,48,and 72 h of treatment,the cell proliferation ability was detected by CCK-8.The foam formation was observed by oil red O staining in the cells treated for 72 h in each group.The protein levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),malondialdehyde(MDA),and superoxide dismutase(SOD)in the cell superna‐tant were measured by ELISA.The reactive oxygen species(ROS)content was determined by the 2-hydroxyethylfluoresce‐in oxidation fluorescence reaction.The relative mRNA expression levels of protein tyrosine kinase 2(JAK2)and signal transducer and activator of transcription 3(STAT3)were detected by real-time fluorescence quantitative PCR.The relative protein expression levels of JAK2,p-JAK2,STAT3,and p-STAT3 were determined by Western blotting,and the ratios of p-JAK2/JAK2 and p-STAT3/STAT3 were calculated.Results Compared with the control group,the OD values of the model group decreased at the same time points(all P<0.05).Compared with the model group at the same time points,the OD values of the high-dose Danyu formula group and the atorvastatin group increased,and the OD values of the medium-dose Danyu formula group increased at 48 and 72h(all P<0.05).The control group cells did not exhibit foam formation,while the cells in the model group showed obvious foam formation.Compared with the model group,the foam formation i

关 键 词：胆瘀方 内皮细胞损伤 脂质泡沫化 炎症反应 JAK2/STAT3通路 动脉粥样硬化 

分 类 号：R543.5[医药卫生—心血管疾病]