地龙提取物减弱矽尘诱导小鼠肺纤维化作用与a-SMA、TGF-β表达关系研究  

Research on Effcet of Dilong(Pheretima)Extract on Attenuating Silicious Dust Induced Pulmonary Fibrosis in Mice and Its Relationship with Expression ofα-SMA,TGF-β

作　　者：宋倩男 张明浩 李可心 邓琳千 李淼 冯大庆 李贵星 陈宏[1] SONG Qiannan;ZHANG Minghao;LI Kexin;DENG Linqian;LIMiao;FENG Daqing;LI Guixing;CHEN Hong(First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150040,Heilongjiang,China)

机构地区：[1]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040

出　　处：《辽宁中医药大学学报》2022年第8期24-27,共4页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：中国博士后科学基金(2020T130178);黑龙江省博士后科研启动金资助项目(LBH-Q20178);黑龙江省自然科学基金项目(H2018052);黑龙江省中医药科研项目(ZHY19-004)。

摘　　要：目的研究地龙提取物减弱矽尘诱导的小鼠肺纤维化作用及与a-SMA、TGF-β表达的关系。方法选取的SPF BALB/c小鼠进行常规饲养,3组小鼠每组各20只采用随机数字表法将患矽尘病小鼠随机分组,正常鼠注射生理盐水作为空白组,矽肺鼠气管注射等量生理盐水作为模型组,矽肺鼠腹腔注射地龙提取物作为地龙提取物组,测定并记录小鼠0~5周内体质量具体变化情况,采用染色试剂染色后显微镜下观察肺纤维细胞增殖速率状况。采用RT-PCR荧光标记法检测TGF-β1及α-SMA mRNA的表达采用免疫荧光染色检测TGF-β1及α-SMA蛋白表达结果,探讨地龙提取物减弱矽尘诱导的小鼠肺纤维化作用及与a-SMA、TGF-β表达的关系。结果空白组小鼠od值为(0.328±0.0045),其肺纤维细胞增殖水平最低,模型组小鼠od值为(0.411±0.0100),其肺纤维细胞增殖水平最高,地龙提取物组小鼠od值为(0.359±0.0050),水平介于空白组与模型组之间,差异有统计学意义(χ2=14.9887,P=0.0006)。0~5周小鼠体质量呈现出增长趋势,第1、3、5周模型组小鼠体质量与空白组、地龙提取物组小鼠体质量比较有明显差异,差异有统计学意义(P<0.05)。小鼠肺纤维α-SMA蛋白表达水平空白组(1.000±0.019)明显低于模型组(1.739±0.101),模型组(1.739±0.101)明显高于地龙提取物组(1.463±0.047),比较存在差异且具有统计学意义(P<0.05)。TGF-β1蛋白表达水平,地龙提取物组(1.295±0.042)较模型组(1.521±0.091)明显偏低,比较存在差异且具有统计学意义(P<0.05)。空白组α-SMA、TGF-β1 mRNA相对表达量较模型组低,差异有统计学意义(P<0.05);经地龙提取物干预后,地龙提取物α-SMA、TGF-β1 mRNA相对表达量较模型组α-SMA、TGF-β1 mRNA相对表达量显著降低,差异有统计学意义(P<0.05)。结论研究结果表明地龙提取物可明显降低肺纤维化组织细胞基质外聚集与沉积的细胞质基质,能调控TGF-β1蛋白与α-SMA蛋�Objective To investigate the effect of Dilong(Pheretima)extract on attenuating silica dustinduced lung fibrosis in mice and its relationship with the expression ofα-SMA and TGF-β.Methods SPF BALB/c mice were selected for routine feeding.Three groups of mice in each group of 20 random number table method of silicosis mice were randomly grouped.The normal mice were injected with normal saline as a blank group.The trachea of silicosis rats were injected the same amount of normal saline as the model group,and the silicosis rats were intraperitoneally injected with the Dilong(Pheretima)extract as the observation group.The specific changes of the mice’s body weight within 0-5 weeks were measured and recorded.The proliferation rate of the lung fiber cells was observed under the microscope after staining with staining reagent.The expression of TGF-β1 andα-SMA mRNA was detected by RT-PCR fluorescence labeling method.The expression of TGF-β1 andα-SMA protein was detected by immunofluorescence staining,and the effect of Dilong(Pheretima)extract on silicious dust induced pulmonary fibrosis in mice and the relationship withα-SMA,TGF-βexpression.Results In the blank group,the od value of the mice was(0.328±0.0045),and the lung fiber cell proliferation level was the lowest.In the model group,the od value was(0.411±0.0100),and the lung fiber cell proliferation level was the highest.In the Dilong(Pheretima)extract group,the od value was(0.359±0.0050),the level was between the blank group and the model group,the difference was statistically significant(χ~2=14.9887,P=0.0006).The weight of the mice showed an increasing trend from 0-5 weeks.The body weight of the model group at week 1,3,and 5 was significantly different from that of the blank group and the Dilong(Pheretima)extract group,and the difference was statistically significant(P<0.05).The expression level ofα-SMA protein in mouse pulmonary fibrosis in the blank group was(1.000±0.019)significantly lower than the model group(1.739±0.101),and the model group(1.7

关 键 词：地龙提取物 矽肺 肺纤维化 A-SMA TGF-Β 

分 类 号：R285.5[医药卫生—中药学]