中国中药杂志

2019, v.44(24) 5473-5478

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地龙对肺纤维化小鼠肺组织中纤维化因子TGF-β1及α-SMA表达的影响
Effect of Dilong on expression of fibrogenic factors TGF-β1 and α-SMA in lung tissue of mice with pulmonary fibrosis

王慧慧;蒙艳丽;杨志敏;王晓溪;徐慧星;王伟明;
WANG Hui-hui;MENG Yan-li;YANG Zhi-min;WANG Xiao-xi;XU Hui-xing;WANG Wei-ming;Heilongjiang Academy of Chinese Medicine Sciences;

摘要(Abstract):

探究地龙对博来霉素诱导的肺纤维化小鼠纤维化因子TGF-β1及α-SMA表达的影响。通过Biacore技术初步检测地龙与纤维化因子TGF-β1的结合力,并采用体内药效学加以验证。将60只雌雄各半的C57小鼠随机分为空白组,模型组,地龙高、中、低剂量组,醋酸泼尼松组;除空白组外(予以气管内注射0. 08 m L·kg-1氯化钠注射液),其余5组予以气管内注射博来霉素4 mg·kg-1来复制肺纤维化模型。药物治疗14 d后,采用Masson染色、免疫组化、RT-PCR技术检测TGF-β1及α-SMA的表达。Biacore实验结果显示,地龙提取液在体外与TGF-β1蛋白有较好的结合,结合值达到619. 3; Masson结果显示,与模型组相比,地龙高、中、低剂量组纤维素沉积均有不同程度减轻; RT-PCR结果显示,不同剂量的地龙提取液在一定程度上均能降低TGF-β1和α-SMA蛋白及mRNA的表达,且呈现一定的剂量依赖性。综上所述,地龙可通过与靶蛋白TGF-β1结合从而抑制α-SMA的表达来延缓肺纤维化进程。
The aim of this paper was to investigate the effect of Dilong( geosaurus) on the expressions of fibrotic factors TGF-β1 and α-SMA in bleomycin-induced pulmonary fibrosis mice. The binding ability of Dilong to fibrotic factor TGF-β1 was initially detected by Biacore technology and verified by in vivo pharmacodynamics. A total of 60 SPF C57 mice were randomly divided into 6 groups. Except the blank group( injecting 0. 08 m L·kg-1 sodium chloride in the trachea),the other five groups were given bleomycin( 4 mg·kg-1) to replicate the pulmonary fibrosis model. After 14 days of drug treatment,the expressions of TGF-β1 and α-SMA were detected by Masson staining,immunohistochemistry and RT-PCR. The results of Biacore experiment showed that the extract of Dilong was well bound to TGF-β1 protein in vitro,and the binding value reached 619. 3. Compared with the model group,Masson's results showed that cellulose deposition in high-dose,medium-dose and low-dose Dilong groups decreased to varying degrees. RT-PCR results showed that different doses of Dilong could reduce protein and mRNA expressions of TGF-β1 and α-SMA to a certain extent in a dose-dependent manner. In conclusion,Dilong could delay the process of pulmonary fibrosis by binding to target protein TGF-β1 and inhibiting the expression of α-SMA.

关键词(KeyWords): 地龙;肺纤维化;Biacore技术;TGF-β1;α-SMA
Dilong(geosaurus);pulmonary fibrosis;Biacore technology;TGF-β1;α-SMA

Abstract:

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基金项目(Foundation): 国家自然科学基金青年基金项目(81603367);国家自然科学基金面上项目(81374045);; 黑龙江省自然科学基金联合引导项目(LH2019H089)

作者(Author): 王慧慧;蒙艳丽;杨志敏;王晓溪;徐慧星;王伟明;
WANG Hui-hui;MENG Yan-li;YANG Zhi-min;WANG Xiao-xi;XU Hui-xing;WANG Wei-ming;Heilongjiang Academy of Chinese Medicine Sciences;

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