摘要
目的 研究炎症促进淋巴管生长的机制。方法 对BALB/c小鼠行角膜缝合手术,建立模型,模拟炎症过程。取正常小鼠和炎症模型小鼠眼球,采用角膜冰冻切片和免疫荧光染色方法定位核因子κB (NF-κB)在生理和炎性淋巴管内皮上的表达。再用Western blot对角膜组织进行NF-κB表达的蛋白定量分析;对炎性模型小鼠术后皮下注射NF-κB的特异性抑制剂MG-132,根据注射与否将小鼠分为治疗组(n=10)和对照组(n=10),采用逆转录荧光定量PCR法检测两组小鼠角膜血管生成素(Ang)-2 mRNA的表达,同时进行HE染色和免疫组织化学染色对比两组小鼠淋巴管生长情况。结果 免疫荧光染色显示,与生理淋巴管相比,炎性淋巴管内皮细胞NF-κB表达较高;与正常小鼠角膜基质相比,炎性角膜NF-κB蛋白表达水平逐渐升高;MG-132治疗组Ang-2 mRNA的表达在注射后第1天达到高峰,随后逐渐下降,而对照组Ang-2 mRNA的表达水平逐渐升高(P<0.01);正常角膜仅在角膜缘有淋巴管生长,在MG-132注射后14 d,与对照组相比,治疗组的淋巴管数量明显减少(P<0.01),管径更细小。结论 炎性环境下,NF-κB是Ang-2的启动子,其表达上调可诱导Ang-2蛋白合成的增加,最终激发并促进炎性淋巴管的生长。
Objective To investigate the mechanism of inflammation promoting lymphatic growth. Methods BALB/c mice were sutured to simulate the inflammatory process. The expression of nuclear factor kappa-B(NF-κB) in physiological and inflammatory lymphatic endothelium was located by corneal frozen section and immunofluorescence staining. Then, the protein expression of NF-κB in corneal tissue was quantitatively analyzed by Western blot. The inflammatory model mice were subcutaneously injected with MG-132, a specific proteasome inhibitor of NF-κB, after operation, and the mice were divided into treatment group(n=10) and control group(n=10) according to whether they were injected or not. The expression of Ang-2 mRNA in cornea of the two groups was detected by reverse transcription fluorescence quantitative PCR, and the growth of lymphatic vessels of the two groups was compared by HE staining and immunohistochemical staining. Results Immunofluorescence staining showed that the expression of NF-κB in inflammatory lymphatic endothelial cells was higher than that in physiological lymphatic. Compared with normal mouse corneal stroma, the expression level of NF-κB protein in inflammatory cornea was higher. The expression of Ang-2 mRNA in MG-132 treatment group reached the peak on the first day after injection, and then decreased gradually, while the expression level of Ang-2 mRNA in control group increased gradually(P<0.01). In normal cornea, lymphatic vessels grew only in the limbal. Fourteen days after MG-132 injection, compared with the control group, the number of lymphatic vessels in the treatment group were significantly reduced(P<0.01) and the diameter of lymphatic ressels was smaller. Conclusion In inflammatory environment, NF-κB is the promoter of Ang-2. It’s up-regulated expression induces the increase of Ang-2 protein synthesis, and finally stimulates and promotes the growth of inflammatory lymphatic vessels.
作者
许晋升
王怡
严志新
XU Jin-sheng;WANG Yi;YAN Zhi-xin(Department of Burn and Plastic Surgery,Affiliated Hospital of Jiangsu University,Zhenjiang,Jiangsu 212000,China)
出处
《中国临床研究》
CAS
2022年第3期297-302,共6页
Chinese Journal of Clinical Research
基金
国家自然科学基金(81641174)。
