摘要
目的:探讨褪黑素对博来霉素诱导的肺纤维化小鼠α-SMA和E-cadherin表达的影响及其机制。方法:用博来霉素(5.0mg/kg)气管内注射制作小鼠肺纤维化模型。随机分为模型组和褪黑素组。4周后处死小鼠提取肺组织。HE染色评价肺组织病理形态改变,氧化应激试剂盒检测超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量;RT-PCR检测肺组织中炎症因子IL-6、TNF-α水平;免疫组织化学检测肺组织中α-SMA和E-cadherin表达;RT-PCR检测肺组织中α-SMA和E-cadherinm RNA的表达水平;Westernblot检测肺组织中α-SMA和E-cadherin的蛋白表达水平。结果:HE结果显示,与模型组比,褪黑素组小鼠肺组织肺泡的纤维化性变较少;与模型组比,褪黑素组小鼠肺组织SOD活性显著升高(P<0.05),MDA表达显著降低(P<0.05);肺组织TNF-α、IL-6表达量显著降低(P<0.05);免疫组织化学结果显示与模型组比,褪黑素组小鼠肺组织α-SMA蛋白表达量显著降低,E-cadherin蛋白表达量显著升高(P<0.05);RT-PCR实验结果显示与模型组相比,褪黑素组小鼠肺组织α-SMAm RNA表达量显著降低(P<0.05),E-cadherinm RNA表达量显著升高(P<0.05);Westernblot结果发现,与模型组比,褪黑素组小鼠肺组织α-SMA蛋白表达量显著降低(P<0.05),E-cadherin蛋白表达量显著升高(P<0.05)。结论:褪黑素通过降低氧化应激和炎症因子水平,抑制α-SMA蛋白表达,促进E-cadherin蛋白表达,延缓肺纤维化进展。
Objective： To investigate the effect of melatonin on expression of α-SMA and E-cadherin in mice with bleomycin induced pulmonary fibrosis and its mechanism. Methods： Mice pulmonary fibrosis model was made by intratracheal injection of bleomycin（5.0 mg/kg） and randomly divided as model group and melatonin group. After 4 weeks, the mice were sacrificed and lung tissue extracted. HE staining was used to evaluate the morphological changes and the oxidative stress kit was used to detect the activity of superoxide dismutase（SOD） and the content of malondialdehyde（MDA）. The levels of inflammatory cytokines IL-6 and TNF-α in lung tissues were measured by RT-PCR. The expression of α-SMA and E-cadherin in lung tissue was detected by immunohistochemistry. The expression of α-SMA and E-cadherin mRNA in lung tissue was detected by RTPCR.Western blot was used to detect the expression of α-SMA and E-cadherin in lung tissue. Results： Compared with the model group, the SOD expression in the melatonin group was significantly increased（P〈0.05）, and the MDA expression was significantly decreased（P〈0.05）; the expression of TNF-α and IL-6 in the lung tissue was significantly decreased（P〈0.05）. Immunohistochemistry results showed that the expression of α-SMA protein in the lung tissue of melatonin group was significantly reduced（P〈0.05）, and E-cadherin protein expression was significantly increased（P〈0.05）; RT-PCR experiment results showed that the expression of α-SMA mRNA in the lungs of melatonin mice was significantly decreased（P〈0.05）, and E-cadherin mRNA expression was significantly increased（P〈0.05）. Western blot results showed that the expression of α-SMA protein in lung tissue of melatonin mice was significantly decreased（P〈0.05）, and E-cadherin protein expression was significantly increased（P〈0.05）. Conclusion： Melatonin can inhibit the expression of α-SMA protein, reduce the expression of E-cadherin, and delay the progression of pulmonary fibr
作者
胡青青
周恩敏
金克
杜勇
钱燕
翁华春
HU Qingqing, ZHOU Enmin, JIN Ke, DU Yong, QIAN Yan, WENG Huachun.(Department of Pediattics, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 32501)
出处
《温州医科大学学报》
CAS
2018年第8期577-581,共5页
Journal of Wenzhou Medical University
基金
温州市科技计划项目(Y20160225)
