Gao Yun-Hang, Song Ling, Chen Teng-Fei, Zheng Zhi-Yuan, Yang Yi-Fei, Yan Can-Mei, Zhang Guang-Ping, Li Han
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
Guangxi Wuzhou Pharmaceutical (Group) Co., Ltd. Wuzhou 543000, China Guangxi Key Laboratory of Notoginseng Comprehensive Utilization Technology Wuzhou 543000, China.
Zhongguo Zhong Yao Za Zhi. 2024 Aug;49(16):4313-4320. doi: 10.19540/j.cnki.cjcmm.20240318.703.
This study aims to investigate the mechanism of Xueshuantong Injection(XST) on pulmonary fibrosis induced by bleomycin(BLM) in rats based on the coagulation cascade pathway. Sixty SD rats were randomly divided into sham surgery group,model group, pirfenidone(PFD, 50 mg·kg(-1)) group, and 27, 54, and 81 mg·kg(-1) XST groups. The rat model of pulmonary fibrosis was established by intratracheal injection of BLM(5 mg·kg(-1)). After 24 hours, the administration groups were given corresponding drugs, while the sham surgery group and model group were given equal volumes of saline. On the 28th day, samples were collected,and the imaging and collagen fiber changes in the lungs of rats were observed. Immunofluorescence(IF) method was used to detect the expression level of alpha-smooth muscle actin(α-SMA), collagen Ⅰ(Col-Ⅰ), E-cadherin(E-cad), and vimentin(Vim). Western blot was used to determine the protein expression of α-SMA, Col-Ⅰ, Vim, and E-cad. Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of prothrombin fragment(F1 + 2), thrombin-antithrombin complex(TAT), soluble fibrin monomer complex(SFMC), and rat fibrinogen degradation products(FDP) in rat lung tissue. Finally, the mRNA and protein levels of protease activated receptor 1(PAR-1) were detected by RT-qPCR, western blot, and IF. Compared with the model group, the scanning of the lungs of rats receiving XST treatment also exhibited patchy and non-homogeneous shadows, but these shadows were less dense than those in the model group. At the same time, there was a significant decrease in Col-Ⅰ fibers in the lungs of rats, and XST could inhibit epithelial-mesenchymal transition(EMT) and downregulate α-SMA and Col-Ⅰ protein expression. In the aspect of the coagulation system, administration of 81 mg·kg(-1) XST significantly reduced the levels of SFMC and FDP. Meanwhile, 81 mg·kg~(-1) XST significantly downregulated the mRNA and protein levels of PAR-1. XST has an anti-pulmonary fibrosis effect in rats, and its mechanism may be related to the downregulation of PAR-1 to rebalance the coagulation cascade pathway.
本研究旨在基于凝血级联途径探讨血栓通注射液（XST）对博来霉素（BLM）诱导的大鼠肺纤维化的作用机制。将60只SD大鼠随机分为假手术组、模型组、吡非尼酮（PFD，50 mg·kg⁻¹）组以及27、54和81 mg·kg⁻¹ XST组。通过气管内注射BLM（5 mg·kg⁻¹）建立大鼠肺纤维化模型。24小时后，给药组给予相应药物，假手术组和模型组给予等体积生理盐水。在第28天采集样本，观察大鼠肺组织的影像学及胶原纤维变化。采用免疫荧光（IF）法检测α-平滑肌肌动蛋白（α-SMA）、Ⅰ型胶原（Col-Ⅰ）、E-钙黏蛋白（E-cad）和波形蛋白（Vim）的表达水平。采用蛋白质印迹法测定α-SMA、Col-Ⅰ、Vim和E-cad的蛋白表达。采用酶联免疫吸附测定（ELISA）法检测大鼠肺组织中凝血酶原片段（F1 + 2）、凝血酶-抗凝血酶复合物（TAT）、可溶性纤维蛋白单体复合物（SFMC）和大鼠纤维蛋白原降解产物（FDP）的水平。最后，通过实时定量聚合酶链反应（RT-qPCR）、蛋白质印迹法和IF检测蛋白酶激活受体1（PAR-1）的mRNA和蛋白水平。与模型组相比，接受XST治疗的大鼠肺部扫描也显示出斑片状和不均匀阴影，但这些阴影比模型组的密度小。同时，大鼠肺组织中Col-Ⅰ纤维显著减少，XST可抑制上皮-间质转化（EMT）并下调α-SMA和Col-Ⅰ蛋白表达。在凝血系统方面，给予81 mg·kg⁻¹ XST可显著降低SFMC和FDP水平。同时，81 mg·kg⁻¹ XST可显著下调PAR-1的mRNA和蛋白水平。XST对大鼠具有抗肺纤维化作用，其机制可能与下调PAR-1以重新平衡凝血级联途径有关。
