摘要
目的探讨乳脂球表皮生长因子8(MFG-E8)抑制大鼠深静脉血栓形成(DVT)及对Toll样因子受体4(TLR4)/核因子-κB(NF-κB)信号通路的影响。方法健康雄性SD大鼠90只, 随机数字表法分为假手术组、模型组、MFG-E8组, 每组30只, Reyers法制备大鼠DVT模型, MFG-E8组大鼠每日给予rhMFG-E8 20 μg/kg尾静脉注射, 连续7 d, 假手术组和模型组大鼠采取尾静脉注射等量生理盐水。采用苏木精-伊红(HE)染色观察下腔静脉血栓形成及内容物组织病理学变化;蛋白质印迹法(Western blot)检测TLR4/NF-κB信号通路相关蛋白细胞TLR4、NB-κBp65(p65)表达, 酶联免疫吸附法(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-4、IL-10表达水平。组间比较采用t检验。结果模型组可见腔静脉内广泛血栓形成, 且随时间延长逐渐增高, 第3天达到高峰, 血栓湿重[(23.6±2.42) mg], 干重[(4.32±0.39) mg], 而MFG-E8组腔静脉内血栓明显减少, 第3天血栓湿重[(14.37±1.85) mg], 干重[(3.02±0.27) mg], 与模型组比较, 差异有统计学意义(t=9.582、8.667, P<0.05);模型组TLR4表达术后第3天达高峰(1.39±0.15), MFG-E8组TLR4表达显著低于模型组(0.51±0.03, t=18.192, P<0.05);模型组NF-κBp65表达术后第3天达高峰(1.25±0.13), MFG-E8组NF-κB p65表达显著低于模型组(0.57±0.04, t=15.810, P<0.05);模型组TNF-α表达术后第3天达高峰[(358.1±12.36) pg/ml], MFG-E8组TNF-α表达显著低于模型组[(205.12±7.33) pg/ml, t=33.665, P<0.05];模型组IL-1β表达术后第3天为(52.73±5.33) pg/ml, MFG-E8组IL-1β表达显著低于模型组[(30.14±2.97) pg/ml, t=11.708, P<0.05];模型组IL-4表达术后第3天达低峰[(28.11±4.34) pg/ml], MFG-E8组IL-4表达显著高于模型组[(48.23±4.90) pg/ml, t=9.720, P<0.05];模型组IL-10表达术后第3天达低峰[(13.82±2.53) pg/ml], MFG-E8组IL-10表达显著高于模型组[(30.87±3.42) pg/ml, t=12.674, P<0.05]。结论 MFG-E8可能通过下调TLR4/NF-κB信号通路, 降低�
Objective To observe the inhibitory effects of milk fat globule epidermal growth factor 8(MFG-E8)on deep venous thrombosis(DVT)in rats and effects on Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway.Methods Totally,90 male SD rats were randomly divided into sham operation group,DVT model group,and MFG-E8 group(n=30 in each group).The DVT model was established by Reyers methods in model group and MFG-E8 group.Rats in sham operation group were simply subjected to the open and suture of the abdominal cavity.Rats in MFG-E8 group were given MFG-E820μg/kg via the tail vein for 7 consecutive days,while the DVT model group and sham operation group were injected with the same amount of normal saline.The pathological changes of venous thrombosis were observed by hematoxylin-eosin(HE)staining.The expression levels of TLR4 and NF-κB p65 were detected by Western blotting,and those of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-4 and IL-10 were detected by enzyme linked immunosorbent assay(ELISA).T-test was used for comparison between groups.Results Extensive intraluminal thrombosis was observed in the model group,and gradually increased with time,reaching the peak on day 3,with wet weight of thrombosis[(23.6±2.42)mg]and dry weight of thrombosis[(4.32±0.39)mg],while the thrombus in the MFG-E8 group decreased significantly.On day 3,the wet weight and dry weight of thrombus were(14.37±1.85)mg and(3.02±0.27)mg,respectively,which were significantly lighter than those of model group(t=9.582,8.667,P<0.05).The TLR4 expression in the model group peaked on the third day after DVT procedure(1.39±0.15),and significantly decreased in the MFG-E8 group as compared with the model group(0.51±0.03,t=18.192,P<0.05).The expression of NF-κBp65 in the model group peaked on the 3rd day after DVT procedure(1.25±0.13),and significantly decreased in the MFG-E8 group as compared with the model group(0.57±0.04,t=15.810,P<0.05).The expression of TNF-αin the model group reached the peak on the third day after
作者
刘青云
黄振华
刘亚坤
唐飞飞
高思海
Liu Qingyun;Huang Zhenhua;Liu Yakun;Tang Feifei;Gao Sihai(Division of Cardiothoracic and Vascular Surgery,Heart-Lung Transplantation Center,Sino-Swiss Heart-Lung Transplantation Institute,Key Laboratory of Organ Transplantation,Ministry of Health,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第2期250-253,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金面上项目(81973992)。
