摘要
目的基于Toll样受体4(Toll-like receptor 4,TLR4)、核因子-κB(nuclear factor-kappa B,NF-κB)蛋白表达探究血府逐瘀合剂对内皮损伤大鼠血管重构及血浆代谢的作用机制。方法选取60只健康清洁级大鼠,将其分为正常组、模型组、血府逐瘀合剂组、辛伐他丁组、TLR4抑制剂组和NF-κB抑制剂组,10只/组,除正常组外均建立血管内皮损伤大鼠模型,血府逐瘀合剂组采用血府逐瘀合剂水煎液进行3 mL/d灌胃,辛伐他丁组采用3 mL/d辛伐他丁进行腹腔注射,TLR4抑制剂组采用3 mg/kg腹腔注射TLR4抑制剂(TAK-242)溶液,NF-κB抑制剂组采用腹腔注射NF-κB抑制剂(PDTF)溶液3 mg/d,模型组与正常组每日采用蒸馏水3 mL灌胃处理,采用ELISA检测血清中白介素1(interleukin-1,IL-1β)、白介素6(interleukin-6,IL-6)、内皮生长因子(vascular endothelial gowth factor,VEGF)水平,采用Image pro-Plus 8.0图像分析软件进行各组血管壁面积、管腔面积、血管总面积计算,采用Western blot检测TLR4、NF-κB蛋白,免疫组化检测单核细胞趋化因子-1(monocyte chemoattractant protein-1,MCP-1)蛋白表达,免疫组化检测单核细胞趋化因子-1(monocyte chemotactic protein-1,MCP-1)水平。结果与正常组比较,模型组IL-1β、IL-6表达升高,VEGF表达降低(P<0.05);与模型组比较,血府逐瘀合剂组IL-1β、IL-6表达降低,VEGF表达升高(P<0.05);与血府逐瘀合剂组比较,辛伐他丁组IL-1β、IL-6表达升高,VEGF表达降低(P<0.05);血府逐瘀合剂组、TLR4抑制剂组、NF-κB抑制剂组组间比较差异无统计学意义(P>0.05)。正常组血管壁内膜完整,无脱落现象,中膜、外膜及血管周围组织均无明显变化,模型组血管外膜厚度明显增加,血管内膜增生严重且面积较大,血府逐瘀合剂组、TLR4抑制剂组、NF-κB抑制剂组内膜面积与外膜厚度明显减小,外膜增生程度降低,辛伐他丁组血管中膜厚度、面积均有轻度减小。与正常组比较,模型组大鼠血管壁面积、管腔面积、血管总面积升高(P<0.05);与模型组比较,血府逐瘀合剂组血管壁面积、管腔面积、血管总面积降低(P<0.05);与血府逐瘀合剂组比较,辛伐他丁组血管壁面积、管腔面积、血管总面积升高(P<0.05);血府逐瘀合剂组、TLR4抑制剂组、NF-κB抑制剂组组间比较差异无统计学意义(P>0.05)。与正常组比较,模型组大鼠血管内TLR4、NF-κB蛋白表达升高(P<0.05);与模型组比较,血府逐瘀合剂组血管内TLR4、NF-κB蛋白表达降低(P<0.05);与血府逐瘀合剂组比较,辛伐他丁组血管内TLR4、NF-κB蛋白表达升高(P<0.05);血府逐瘀合剂组、TLR4抑制剂组、NF-κB抑制剂组组间比较差异无统计学意义(P>0.05)。与正常组比较,模型组大鼠血管内MCP-1吸光度表达升高(P<0.05);与模型组比较,血府逐瘀合剂组血管内MCP-1吸光度表达降低(P<0.05);与血府逐瘀合剂组比较,辛伐他丁组血管内MCP-1吸光度表达升高(P<0.05);血府逐瘀合剂组、TLR4抑制剂组、NF-κB抑制剂组组间比较差异无统计学意义(P>0.05)。结论血府逐瘀合剂对于降低大鼠血清中IL-1β、IL-6水平,提高VEGF因子水平,减轻内皮损伤大鼠的血管壁面积、管腔面积、血管总面积,降低血管内TLR4、NF-κB蛋白表达,降低大鼠血管内MCP-1吸光度等均具有一定积极影响。
Objective To explore the expression of TLR4 and NF-κB protein and the mechanism of Xuefu Zhuyu Mixture(血府逐瘀合剂)on vascular remodeling and plasma metabolism in endothelial injury rats.Methods Selecting 60 rats health clean level only,it can be divided into normal group,model group,Xuefu Zhuyu Mixture group,simvastatin butyl group,TLR4 inhibitor group and NF-κB inhibitor group,each group 10 rats,in addition to the normal group were established vascular endothelial damage in rats model,Xuefu Zhuyu Mixture group uses Xuefu Zhuyu Mixture water decoction for 3 mL/d lavage,simvastatin butyl group with simvastatin butyl 3 mL/d intraperitoneal injection,TLR4 inhibitor group was intraperitoneally injected TAK-242 solution 3 mg/kg,NF-κB inhibitor group was intraperitoneally injected PDTF 3 mg/d,model group and normal group were intragastric treated with distilled water 3 mL daily.Interleukin-1(IL-1β),interleukin-6(IL-6)and endothelial growth factor(VEGF)in serum were detected by ELISA,and the vascular wall area,lumen area and total vascular area were calculated by Image Pro-Plus 8.0 Image analysis software.TLR4 and NF-κB protein were detected by Western blot,and monocyte chemokine-1(MCP-1)was detected by immunohistochemistry.Results Compared with normal group,the expressions of IL-1 and IL-6 in model group were higher,and the expression of VEGF was lower(P<0.05).Compared with model group,the expressions of IL-1 and IL-6 were decreased and the expression of VEGF was increased in Xuefu Zhuyu Mixture group(P<0.05).Compared with Xuefu Zhuyu Mixture group,the expressions of IL-1 and IL-6 in simvastatin group were increased,and the expression of VEGF was decreased(P<0.05).There was no difference between Xuefu Zhuyu Mixture group,TLR4 inhibitor group and NF-κB inhibitor group(P>0.05).There was no abnormal change in the vascular wall tissue in the normal group,the vascular outer membrane thickness increased significantly in the model group,and the intima hyperplasia was serious.The intima area and the intima thickness decreased significantly in the Xuefu Zhuyu Mixture group,TLR4 inhibitor group and NF-κB inhibitor group,and the thickness and the area of the vascular middle membrane decreased slightly in the simvastatin group.Compared with normal group,the expressions of vascular wall area,lumen area and total vascular area in model group were higher(P<0.05).Compared with model group,the above indexes in Xuefu Zhuyu Mixture group were decreased(P<0.05).Compared with Xuefu Zhuyu Mixture group,the above indexes in simvastatin group were increased(P<0.05).There was no difference between Xuefu Zhuyu Mixture group,TLR4 inhibitor group and NF-κB inhibitor group(P>0.05).Compared with normal group,the expressions of TLR4 and NF-κB protein in blood vessels of rats in model group were higher(P<0.05).Compared with model group,the above indexes in Xuefu Zhuyu Mixture group were decreased(P<0.05).Compared with Xuefu Zhuyu Mixture group,the above indexes in simvastatin group were increased(P<0.05).There was no difference between Xuefu Zhuyu Mixture group,TLR4 inhibitor group and NF-κB inhibitor group(P>0.05).Compared with normal group,MCP-1 optical density expression in blood vessels of rats in model group was higher(P<0.05).Compared with model group,the above index in Xuefu Zhuyu Mixture group was decreased(P<0.05).Compared with Xuefu Zhuyu Mixture group,the above index in simvastatin group was increased(P<0.05).There was no difference between Xuefu Zhuyu Mixture group,TLR4 inhibitor group and NF-κB inhibitor group(P>0.05).Conclusion Xuefu Zhuyu Mixture has positive effects on reducing serum IL-1βand IL-6,increasing VEGF factor,reducing vascular wall area,lumen area and total vascular area,reducing the expression of TLR4 and NF-κB protein,and reducing the optical density of MCP-1 in rats with endothelial injury.
作者
肖遥
周健华
胡有志
XIAO Yao;ZHOU Jianhua;HU Youzhi(Hubei Hospital of Traditional Chinese Medicine,Wuhan 430061,Hubei,China)
出处
《辽宁中医药大学学报》
CAS
2023年第4期25-29,共5页
Journal of Liaoning University of Traditional Chinese Medicine
基金
湖北省卫生健康委员会中医药科研项目(ZY2021M047)。
