摘要
目的研究组蛋白去乙酰化酶2(HDAC2)在良性气管狭窄动物模型中的表达,并探讨HDAC2在气管狭窄发生中的作用机制,为治疗良性气管狭窄寻求新靶标。方法将18只家兔完全随机分成空白对照组、模型组和红霉素组,每组各6只。模型组和红霉素组家兔行气管切开并用尼龙刷来回刷20余次损伤气管内壁制作良性气管狭窄的动物模型。术前7 d至术后9 d模型组给予生理盐水灌胃,红霉素组给予小剂量红霉素15 mg·kg^(–1)·d^(–1)灌胃,空白对照组不做任何处理。术后第10 d处死各组家兔,切取狭窄段气管,测量气管狭窄度,并取狭窄段肉芽组织提取RNA和蛋白质。用实时荧光定量PCR检测肉芽组织中HDAC2、白细胞介素-6(IL-6)、IL-8的mRNA相对表达量,用蛋白免疫印迹检测肉芽中HDAC2蛋白的相对表达量。结果模型组与空白对照组比较,气管狭窄明显[(84.60±1.14)%比(27.00±6.44)%],HDAC2 mRNA和蛋白表达下调(0.29±0.07比1.00±0.00,0.20±0.02比0.49±0.04),IL-6、IL-8 mRNA表达上调(4.22±0.67比1.00±0.00,162.72±23.23比1.00±0.00)。红霉素组与模型组比较,气管狭窄度减轻[(64.00±12.25)%比(84.60±1.14)%],HDAC2 mRNA和蛋白表达上调(0.42±0.14比0.29±0.07,0.43±0.01比0.20±0.02),IL-6、IL-8 mRNA表达下调(0.72±0.24比4.22±0.67,130.22±7.93比162.72±23.23),差异均具有统计学意义(均P<0.05)。气管狭窄度与HDAC2 mRNA相对表达量呈负相关(Pearson相关系数r=–0.96,P<0.05)。结论良性气管狭窄动物模型中HDAC2表达下调与气管狭窄的发生发展有关。小剂量红霉素具有治疗良性气管狭窄的作用,可能是小剂量红霉素上调HDAC2的表达,进而抑制气管损伤修复时的炎症失调有关。
Objective To investigate the expression of histone deacetylase 2（HDAC2） in animal model of benign tracheal stenosis, and explore the mechanism of HDAC2 in development of tracheal stenosis. Methods Eighteen rabbits were randomly divided into a blank control group, a model group, and an erythromycin group, with 6 rats in each group.The model group and the erythromycin group underwent tracheostomy, the inner wall of trachea was brushed back and forth with a nylon brush for more than 20 times to induce benign tracheal stenosis. From 7 days before surgery to 9 days after surgery, the model group received gavage with saline, the erythromycin group received gavage with low-dose erythromycin in dose of 15 mg·kg–1·d–1, and the control group did not receive any treatment. On the 10 th day after operation, all the rabbits were sacrificed and the trachea was cut to measure the tracheal stenosis. RNA and protein were extracted from the granulation tissue in the stenosis and the relative m RNA expressions of HDAC2, interleukin（IL）-6 and IL-8 in the granulation tissue were detected by real-time fluorescence quantitative PCR. The relative expression of HDAC2 protein was detected by Western blot. Results Compared with the blank control group, the tracheal stenosis in the model group was more obvious [（84.60±1.14）% vs.（27.00±6.44）%], the mRNA and protein expressions of HDAC2 were decreased（0.29±0.07 vs. 1.00±0.00, 0.20±0.02 vs. 0.49±0.04）, the m RNA expressions of IL-6 and IL-8 were up-regulated（4.22±0.67 vs. 1.00±0.00, 162.72±23.23 vs.1.00±0.00）. Compared with the model group, tracheal stenosis in the erythromycin group was relieved [（64.00±12.25）% vs.（84.60±1.14）%], the mRNA and protein expressions of HDAC2 were increased（0.42±0.14 vs. 0.29±0.07, 0.43±0.01 vs. 0.20±0.02）, the mRNA expressions of IL-6 and IL-8 were decreased（0.72±0.24 vs. 4.22±0.67, 130.22±7.93 vs. 162.72±23.23）. All the differences were statistically significant（all P〈0.05）. The Pearson correlation coefficient between tracheal stenosis and HDAC2 mRNA relative expression was –0.96（P〈0.05）.Conclusions The down-regulation of HDAC2 expression in model of benign tracheal stenosis is related to the occurrence and development of tracheal stenosis. The low dose of erythromycin may be used to treat benign tracheal stenosis by up-regulating expression of HDAC2 and thus inhibiting the inflammatory disorder during tracheal injury repair.
作者
甘敬华
甘罗曼
覃恩愿
孟晓燕
黎雨
李文涛
侯长春
周磊
柳广南
GAN Jinghua;GAN Luoman;QIN Enyuan;MENG Xiaoyan;LI Yu;LI Wentao;HOU Changchun;ZHOU Lei;LIU Guangnan(Department of Respiratory Medicine, Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530007, P. R. China;Qinghai University School of Medicine, Xining, Qinghai 810016, P. R. China;Liuzhou Workers Hospital, Liuzhou, Guangxi 545005, P. R. China)
出处
《中国呼吸与危重监护杂志》
CAS
CSCD
北大核心
2018年第3期285-289,共5页
Chinese Journal of Respiratory and Critical Care Medicine
基金
国家自然科学基金(81760001)
关键词
良性气管狭窄
组蛋白去乙酰化酶2
红霉素
感染
炎症
Benign tracheal stenosis
Histone deacetylase 2
Erythromycin
Infection
Inflammation
