摘要
目的观察天降血栓通丸减少动脉粥样硬化(AS)小鼠主动脉斑块形成的作用,并探讨其作用机制。方法取ApoE-/-雄性小鼠62只,通过饲喂高脂饲料建立ApoE-/-小鼠AS模型。造模成功后,采用随机数字表法将小鼠分为模型组、阿托伐他汀组、天降血栓通丸低剂量组、天降血栓通丸高剂量组、天降血栓通丸+脂多糖(LPS)组,每组12只。另取12只C57BL/6小鼠,饲喂普通饲料,作为正常对照组。阿托伐他汀组灌胃400μL阿托伐他汀(1.5 mg/mL)灌胃,天降血栓通丸低剂量组和高剂量组分别灌胃4.5、9 g/mL的天降血栓通丸药液400μL,天降血栓通丸+LPS组给予18 g/mL的天降血栓通丸药液200μL和10 g/mL的LPS 200μL,正常对照组与模型组灌胃200μL生理盐水。连续灌胃8周后处死小鼠取主动脉,采用油红O染色法观察主动脉斑块的形成情况,计算斑块面积占总血管面积的比值。取各组腹主动脉血,使用全自动生化分析仪检测血脂指标总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C),采用ELISA法检测血清炎症因子白细胞介素6(IL-6)、白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)和血清细胞黏附分子单核细胞趋化因子1(MCP-1)、细胞间黏附分子1(ICAM-1)、血管细胞黏附分子1(VCAM-1)水平。采用RT-PCR法检测主动脉组织TLR4/MyD88/NF-κB信号通路相关基因TLR4、MyD88、NF-κB mRNA的相对表达量,Western blotting法检测TLR4、MyD88、NF-κB蛋白表达量。结果与正常对照组比较,模型组主动脉AS斑块面积增加(P<0.05);与模型组比较,阿托伐他汀组和天降血栓通丸低、高剂量组主动脉AS斑块面积减少(P均<0.05);与天降血栓通丸高剂量组比较,天降血栓通丸+LPS组主动脉AS斑块面积增加(P均<0.05)。与正常对照组比较,模型组血清TC、TG、LDL-C、IL-6、IL-1β、TNF-α、MCP-1、ICAM-1、VCAM-1水平均升高(P均<0.05);与模型组比较,阿托伐他汀组和天降血栓通丸低、高剂量组血清上述指标均降低(P均<0.05);与天降血栓通丸高剂量组比较,天降血栓通丸+LPS组上述指标均升高(P均<0.05)。与正常对照组比较,模型组主动脉组织TLR4、MyD88、NF-κB mRNA及蛋白表达量升高(P均<0.05);与模型组比较,阿托伐他汀组和天降血栓通丸低、高剂量组主动脉组织TLR4、MyD88、NF-κB mRNA及蛋白表达量降低(P均<0.05);与天降血栓通丸高剂量组比较,天降血栓通丸+LPS组上述指标均升高(P均<0.05)。结论天降血栓通丸能够抑制AS小鼠主动脉斑块形成,并发挥调节血脂、抑制炎症反应、保护血管内皮功能的作用,高剂量天降血栓通丸具有更显著的效果;其机制可能与抑制TLR4/MyD88/NF-κB信号通路激活有关。
Objective To observe the effect of Tianjiang Xueshuantong pills in reducing aortic plaque formation in mice with atherosclerosis(AS),and to explore its mechanism.Methods Sixty-two ApoE-/-male mice were selected and fed with high-fat diet to establish the ApoE-/-mouse AS models.After the ApoE-/-mouse AS models were successfully established,they were randomly divided into the model group,atorvastatin group,low-dose Tianjiang Xueshuantong pills group,high-dose Tianjiang Xueshuantong pills group,and Tianjiang Xueshuantong pills+lipopolysaccharide(LPS)group,with 12 in each group;another 12 C57BL/6 mice were taken as the normal group.Mice in the atorvastatin group were given intragastric administration of 400μL atorvastatin(1.5 mg/mL);mice in the low-dose and high-dose Tianjiang Xueshuantong pills groups were given 4.5 and 9 g/mL Tianjiang Xueshuantong pills 400μL,respectively;mice in the Tianjiang Xueshuantong pills+LPS group were given 18 g/mL Tianjiang Xueshuantong pills solution 200μL and 10 g/mL LPS 200μL,and mice in the normal control group and model group were given 200μL normal saline intragastrically.After 8 weeks of continuous gavage,the aorta was taken out and was used for oil red O staining to observe the formation of aortic plaques,and then we calculated the ratio of plaque area to the total area of aortic vessels.Blood was taken from the abdominal aorta,and the levels of total cholesterol(TC),triglyceride(TG),and low-density lipoprotein cholesterol(LDL-C)were detected by automatic biochemical analyzer,and Enzyme-linked immunosorbent assay(ELISA)was used to detect serum inflammatory factors interleukin-6(IL-6),interleukin-1β(IL-1β),tumor necrosis factor alpha(TNF-ɑ)and serum cell adhesion molecule monocytes Chemokine 1(MCP-1),intercellular adhesion molecule 1(ICAM-1),and vascular cell adhesion molecule 1(VCAM-1)levels.RT-PCR was used to detect the relative expression levels of TLR4/MyD88/NF-κB signaling pathway-related genes TLR4,MyD88,and NF-κB mRNA in the aortic tissues.Western blotting was used to detect the expression levels of TLR4,MyD88,and NF-κB proteins.Results Compared with the normal group,the aortic AS plaque area increased significantly in the model group(P<0.05).Compared with the model group,the aortic AS plaque area gradually decreased in the atorvastatin group and low-dose and high-dose Tianjiang Xueshuantong pills groups(all P<0.05).Compared with the high-dose Tianjiang Xueshuantong pills group,the aortic AS plaque area increased significantly in the Tianjiang Xueshuantong pills+LPS group(P<0.05).Compared with the normal group,the levels of TC,TG,LDL-C,IL-6,IL-1β,TNF-ɑ,MCP-1,ICAM-1,VCAM-1 significantly increased in the model group(all P<0.05).Compared with the model group,the above indicators decreased in the atorvastatin group and low-dose and high-dose Tianjiang Xueshuantong pill groups(all P<0.05).Compared with the high-dose Tianjiang Xueshuantong pills group,the above indicators increased in the Tianjiang Xueshuantong pills+LPS group(all P<0.05).Compared with the normal group,the expression levels of TLR4,MyD88,and NF-κB mRNA and protein significantly increased in the model group(all P<0.05).Compared with the model group,the expression levels of TLR4,MyD88,and NF-κB mRNA and protein decreased in the atorvastatin group and low-dose and high-dose Tianjiang Xueshuantong pills groups(all P<0.05).Compared with the high-dose Tianjiang Xueshuantong pills group,the above indicators in the Tianjiang Xueshuantong pills+LPS group significantly increased(all P<0.05).Conclusions Tianjiang Xueshuantong pills can inhibit the formation of aortic plaques in AS mice,and play a role in regulating blood lipids,inhibiting inflammatory responses,and protecting vascular endothelial function.High-dose Tianjiang Xueshuantong pills have more significant effects,and its mechanism may be related to inhibiting the activation of TLR4/MyD88/NF-κB signaling pathway.
作者
赵聪娜
韩鹏飞
张宝楠
王居平
ZHAO Congna;HAN Pengfei;ZHANG Baonan;WANG Juping(Department of Nephrology,Beichen District Hospital of Traditional Chinese Medicine,Tianjin 300400,China;不详)
出处
《山东医药》
CAS
2022年第14期25-30,共6页
Shandong Medical Journal
基金
天津市北辰区中医医院科研项目(SHGY-2020038)。
作者简介
第一作者:赵聪娜(1980-),女,硕士,主治医师,主要研究方向为中医治疗肾脏疾病。E-mail:zcna8856@163.com。
