5首活血化瘀方对氧化低密度脂蛋白诱导人脐静脉内皮细胞损伤的保护作用差异及机制  

Difference and mechanism in the protective effects of five promoting circulation and removing stasis formulas on HUVECs induced by ox-LDL

作　　者：郑彩杏 李金霞[1] 周小青[1] 刘旺华[1] ZHENG Caixing;LI Jinxia;ZHOU Xiaoqing;LIU Wanghua(Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学,长沙410208

出　　处：《中华中医药杂志》2025年第3期1086-1091,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82104775);湖南省教育厅科学研究项目(No.23B0357,No.23B0376);湖南中医药大学优秀青年项目(No.Z2023XJYQ04);湖南省中医药科研课题(No.B2024008);中国科协青年托举工程(No.YESS20220451)。

摘　　要：目的:观察5首活血化瘀方含药血清对氧化低密度脂蛋白(ox-LDL)诱导损伤人脐静脉内皮细胞(HUVECs)的保护作用及机制。方法:将72只大鼠随机分为6组:对照组、丹参饮组(18.9g·kg^(-1)·d^(-1))、桃红四物汤组(25.65g·kg^(-1)·d^(-1))、血府逐瘀汤组(34.2g·kg^(-1)·d^(-1))、活络效灵丹组(27g·kg^(-1)·d^(-1))、失笑散组(5.4g·kg^(-1)·d^(-1)),每组12只,雌雄各半,每组灌胃相应复方,浓度为成人等效剂量的5倍,对照组灌胃等量蒸馏水,每日1次,7d后采血以制备含药血清。以50μg/mLox-LDL诱导HUVECs损伤后,以5首方15%含药血清和5μmol/LSP600125处理24h后检测。以CCK-8法检测细胞增殖;流式细胞仪检测细胞凋亡;WesternBlot检测JNK、p-JNK、Bcl-2、Bax、Caspase9、Caspase3蛋白表达水平;Real-timePCR检测JNK、Bcl-2、Bax、Caspase9、Caspase3mRNA表达水平。根据以上指标对5首方的作用进行排序与赋值,以对比作用力差异。结果:与模型组比较,5首活血化瘀方均能显著增强HUVECs细胞活性(P<0.01),减少细胞凋亡(P<0.05,P<0.01),显著下调JNK、Bax、Caspase9、Caspase3蛋白和mRNA表达水平(P<0.01,P<0.05),显著下调p-JNK蛋白表达水平(P<0.01,P<0.05),显著上调Bcl-2蛋白和mRNA表达水平(P<0.01,P<0.05)。结论:5首活血化瘀方均可不同程度的保护受损的HUVECs,其机制可能与调控JNK信号通路相关。Objective:To observe the protective effect and mechanism of five promoting circulation and removing stasis formulas containing serum on oxidized low-density lipoprotein induced human umbilical vein endothelial cells(HUVECs)injury cells.Methods:Seventy-two rats were randomly divided into 6 groups:control group,Danshen Drink group(18.9g·kg^(-1)·d^(-1)).Taohong Siwu Decoction group(25.65g·kg^(-1)·d^(-1)),Xuefu Zhuyu Decoction group(34.2g·kg^(-1)·d^(-1)),Huoluo Xiaoling Pills group(27g·kg^(-1)·d^(-1)),Shixiao Powder group(5.4g·kg^(-1)·d^(-1)),each group has 12 animals,half male and half female,each group is given the corresponding compound by gavage,the concentration is 5 times the adult equivalent dose,the control group is gavage the same amount of distilled water,once a day.After 7 days,blood was collected to prepare medicated serum.Fiftyμg/mL ox-LDL was used to induce HUVECs injury,and treated with 15%medicated serum of five recipes and 5μmol/L SP600125 for 24 h.CCK-8 method was used to detect cell proliferation;flow cytometry was used to detect cell apoptosis.Western Blot method was used to detect the protein expression levels of JNK,p-JNK,Bcl-2,Bax,Caspase9,and Caspase3.Real-time PCR method was used to detect the gene expression levels of JNK,Bcl-2,Bax,Caspase9 and Caspase3.According to the above indexes,the effects of the five recipes are ranked and assigned to compare the differences in the forces.Results:Compared with the model group,all five recipes could enhance the cell activity of damaged HUVECs(P<0.01),inhibit cell apoptosis(P<0.05,P<0.01),and down-regulate the protein and mRNA expression levels of JNK,Bax,Caspase9 and Caspase3(P<0.01,P<0.05),down-regulate the protein expression level of p-JNK(P<0.01,P<0.05),and up-regulate the protein and mRNA expression levels of Bcl-2(P<0.01,P<0.05).Conclusion:Five recipes can protect damaged HUVECs to varying degrees,and its mechanism may be related to the regulation of JNK signaling pathway.

关 键 词：血瘀证 内皮细胞 JNK 丹参饮 血府逐瘀汤 失笑散 活络效灵丹 桃红四物汤 

分 类 号：R285.5[医药卫生—中药学]