摘要
目的:观察蜂毒素对博莱霉素诱导小鼠肺纤维化的干预作用。方法:70只SPF级C57BL/6小鼠被随机分为正常组10只和模型组、地塞米松组、蜂毒素低剂量组、蜂毒素中剂量组、蜂毒素高剂量组各12只。除正常组外,均采用气管穿刺注入博莱霉素(BLM)制备肺纤维化小鼠模型。从术后第1天开始,地塞米松组按3 mg/kg的剂量腹腔注射,蜂毒素低、中、高剂量组分别给予5μg/(kg·d)、10μg/(kg·d)、20μg/(kg·d),对照组和模型组给予等体积生理盐水灌胃,连续2周。分别于第7、第14天处死动物,收集小鼠外周血样本,通过ELISA方法检测血清转化生长因子(TGF-β1)、胶原蛋白I(Collagen I)、胶原蛋白III(Collagen III)、基质金属蛋白酶2(MMP2)和基质金属蛋白酶9(MMP9)的水平。取肺组织进行苏木精-伊红(HE)分析,Masson染色和羟脯氨酸(HYP)评估以观察组织病理学变化和胶原沉积。采用实时荧光定量(Realime PCR)法和蛋白兔疫印迹法(Western blot)观察各组大鼠肺组织TGF-β1、Smad2、Smad3等蛋白和基因的表达变化。结果:与对照组比较,模型组大鼠肺纤维化明显,HYP、TGF-β1、Collagen I、Collagen I的含量升高(P <0. 05),肺组织TGF-β1、Smad2、Smad3蛋白和基因表达升高(P <0. 05);与模型组比较,蜂毒素中高剂量组血清HYP、TGF-β1、Collagen I、Collagen I的含量下降(P <0. 05),肺组织TGF-β1、Smad2、Smad3蛋白和基因表达降低,差异有统计学意义(P <0. 05)。结论:蜂毒素可以减轻博来霉素诱导的小鼠肺纤维化的程度,其机制可能与抑制TGF-β1/Smads通路有关。
Objective: To observe the effects of melittin on inhibiting pulmonary fibrosis induced by bleomycin in rats. Methods:Seventy SPF grade C57 BL/6 rats were randomly divided into the normal group( n = 10) and the model group,dexamethasone group and melittin low dose group,melittin middle dose group and melittin high dose group( n = 12). In addition to the normal group,a rat model of pulmonary fibrosis was prepared by injecting bleomycin( BLM) into the trachea. From the first day after surgery,the dexamethasone group was intraperitoneally injected at a dose of 3 mg/kg,while the low,medium and high doses of melittin were administered to 5,10,and 20 μg/( kg·d) respectively. The control group and the model group were given an equal volume of saline for 2 weeks. The animals were sacrificed on the 7 th and 14 th day,respectively,peripheral blood samples of which were collected.ELISA was used to detect serum transforming growth factor( TGF-β1),collagen I( Collagen I),collagen III( Collagen III),and matrix metalloproteinase 2( MMP2) and the level of matrix metalloproteinase 9( MMP9). Lung tissue was taken for hematoxylin-eosin( HE) analysis,Masson staining and hydroxyproline( HYP) evaluation to observe histopathological changes and collagen deposition.The expressions of TGF-β1,Smad2,Smad3 and other proteins and genes in lung tissue of each group were observed by Real-time PCR and Western blot. Results: Compared with the control group,the pulmonary fibrosis was significantly increased in the model group. The contents of HYP,TGF-β1,Collagen I and Collagen I were increased( P < 0. 05),and the expression of TGF-β1,Smad2,Smad3 protein and gene in lung tissue was increased( P < 0. 05);Compared with the model group,the levels of serum HYP,TGF-β1,Collagen I,Collagen I in the high dose group of melittin decreased( P < 0. 05),and TGF-β1,Smad2,Smad3 protein in lung tissue and gene expression was decreased( P < 0. 05). There was no statistically significant difference in the low-dose group. Conclusion: Melittin can effectively
作者
李莉
危蕾
王众福
张秀莲
钱叶长
Li Li;Wei Lei;Wang Zhongfu;Zhang Xiulian;Qian Yechang(Baoshan Branch,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201900,China)
出处
《世界中医药》
CAS
2019年第3期608-614,共7页
World Chinese Medicine
基金
2016年上海市卫生和计划生育委员会基金项目(20164Y0241)--CXCL14通过TGF-β/Smads通路影响肺纤维化的机制研究
