基于NF-κB通路探讨菲牛蛭对颈动脉损伤兔血管内皮的保护作用  

Hirudinaria manillensis protects vascular endothelium of rabbits with carotid artery injury via NF-κB pathway

作　　者：周豪 张颖[1] 邓淦予 俸一然 郭春霞 杜义斌 张学娅[1] ZHOU Hao;ZHANG Ying;DENG Ganyu;FENG Yiran;GUO Chunxia;DU Yibin;ZHANG Xueya(First Affililated Hospital of Kunming Medical University,Kunming 650000,Yunnan,China;Yunnan University of Chinese Medicine,Kunming 650000,Yunnan,China)

机构地区：[1]昆明医科大学第一附属医院,云南昆明650000 [2]云南中医药大学,云南昆明650000

出　　处：《现代中西医结合杂志》2024年第21期2937-2945,共9页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：2020年度云南省兴滇人才“名医”专项(RLMY20200003);云南省科学技术厅科技计划项目(202101AY070001-097)。

摘　　要：目的基于核因子κB(NF-κB)信号通路,从抗炎角度探讨菲牛蛭对颈动脉球囊损伤兔损伤段血管内皮的保护作用。方法将36只兔随机分为假手术组、模型组、西药组及菲牛蛭低、中、高剂量组,每组6只。假手术组不插入球囊导管,术后生理盐水灌胃和普通饲料喂养;其余组兔建立颈动脉球囊损伤模型,术后给予高脂饲料喂养,其中模型组同时给予生理盐水灌胃,菲牛蛭低、中、高剂量组分别给予0.1 g/(kg·d)、0.4 g/(kg·d)、0.8 g/(kg·d)菲牛蛭灌胃,西药组给予阿司匹林肠溶片125 mg/d、氯吡格雷片62.5 mg/d、阿托伐他汀钙片5 mg/d灌胃,均1次/d,连续灌胃10 d。用空气栓塞法处死兔后,采集血清及颈动脉损伤段血管。采用苏木精-伊红(HE)染色及油红O(ORO)染色观察损伤血管组织病理形态,免疫组化法检测血管组织中NF-κB p65、NOD样受体蛋白3(NLRP3)阳性表达情况,免疫荧光单标法检测血管组织中半胱氨酸天冬氨酸蛋白酶1(Caspase-1)表达情况,Western blot法检测血管组织中NF-κB p65、NLRP3、Caspase-1蛋白表达情况,酶联免疫吸附法检测血清C反应蛋白(CRP)、白细胞介素-1β(IL-1β)和白细胞介素-18(IL-18)水平。结果假手术组兔血管壁光滑,未见增生狭窄和脂质沉积;模型组兔血管内膜明显增生,管腔不同程度狭窄,见大量脂质沉积;与模型组相比,各药物组血管内膜增生和管腔狭窄减轻,脂质沉积减少。模型组兔血管组织中NF-κB p65、NLRP3、Caspase-1蛋白阳性表达面积和蛋白相对表达量及血清CRP、IL-1β、IL-18水平均明显高于假手术组(P均<0.05);各药物组兔血管组织中NF-κB p65、NLRP3、Caspase-1蛋白阳性表达面积和蛋白相对表达量及血清CRP、IL-1β、IL-18水平均明显低于模型组(P均<0.05),且菲牛蛭高剂量组上述各蛋白阳性表达面积、蛋白相对表达量及血清各炎症指标水平均明显低于菲牛蛭低剂量组(P均<0.05)。结论菲牛蛭可�Objective It is to investigate the protective effect of Hirudinaria manillensis on the vascular endothelium of the injured segment of carotid artery balloon injured rabbits from an anti-inflammatory perspective based on the nuclear factorκB(NF-κB)signaling pathway.Methods Thirty-six rabbits were randomly divided into sham operation group,model group,western medicine group,and low,medium,and high dose group of Hirudinaria manillensis,with 6 rabbits in each group.The rabbits in the sham operation group was not inserted with balloon catheter,and were given normal saline by gavage and fed with normal feed after operation;the rabbits in the rest groups were modeled to establish models of carotid artery balloon injury and fed with high-fat feed after operation,at the same time,the model group was given normal saline by gavage,the low,medium,and high dose group of Hirudinaria manillensis were given Hirudinaria manillensis 0.1 g/(kg·d),0.4 g/(kg·d),and 0.8 g/(kg·d)by gavage,respectively,and the western medicine group was given aspirin enteric-coated tablets 125 mg/d,clopidogrel tablets 62.5 mg/d,and atorvastatin calcium tablets 5 mg/d by gavage,all once daily,continuously treated for 10 days.After the rabbits were killed by air embolization,their blood serum and blood vessels from the injured segment of the carotid artery were collected.The histopathological morphology of injured blood vessels were observed by HE staining and ORO staining,the positive expressions of NF-κB p65 and NOD-like receptor protein 3(NLRP3)were detected by immunohistochemistry,the expression of cysteine aspartate protease 1(Caspase-1)in vascular tissues was detected by immunofluorescence single-labeling,the protein expressions of NF-κB p65,NLRP3,and Caspase-1 in vascular tissues were detected by Western blot method,and the serum levels of C-reactive protein(CRP),interleukin-1β(IL-1β),and interleukin-18(IL-18)were detected by enzyme-linked immunosorbent assay.Results The blood vessel wall was smooth with no hyperplasia,stenosis or lipid de

关 键 词：动脉粥样硬化 菲牛蛭 核因子κB通路 炎症反应 NOD样受体蛋白3 

分 类 号：R-332[医药卫生]