Role of Cytoskeletal Proteins in Rat PASMCs Migration Induced by HPS Rat Serum

目的：本研究聚焦于探究骨架蛋白在肝肺综合征(HPS)大鼠血清诱导肺动脉平滑肌细胞(PASMCs)迁移进程中的作用。方法：实验选用20只雄性Sprague-Dawley大鼠，通过慢性胆总管结扎的方法构建大鼠HPS模型，随后采集腹主动脉血用于制备血清。将培养的大鼠PASMCs分别接种至6孔板与24孔板，借助随机数字表法分为两组，即对照组(C组)与HPS组。C组和HPS组分别采用正常大鼠和HPS大鼠血清进行孵育，并使两组血清终浓度均调控至5%。在细胞孵育的24小时(T1)、48小时(T2)以及72小时(T3)这三个时间节点，运用Western-blot技术对PASMCs内的骨架蛋白，包括F-actin、α-tubulin以及Destrin的蛋白表达量进行测定；同时，利用Transwell小室(于T1时间点)以及划痕实验(在T1至T3时间点)检测PASMCs的迁移水平。结果：相较于C组，HPS组的PASMCs迁移能力显著提升，而骨架蛋白(F-actin，α-tubulin及Destrin)的蛋白表达则呈现出明显的下调趋势(P Objective: This study focuses on exploring the role of cytoskeletal proteins in the migration process of pulmonary artery smooth muscle cells (PASMCs) induced by the serum of rats with hepatopulmonary syndrome (HPS). Methods: Twenty male Sprague-Dawley rats were selected for the experiment. A rat model of HPS was established by chronic common bile duct ligation, and then the blood from the abdominal aorta was collected for serum preparation. The cultured rat PASMCs were separately seeded into 6-well plates and 24-well plates, and divided into two groups by the random number table method, namely the control group (Group C) and the HPS group. Group C and the HPS group were incubated with the serum of normal rats and HPS rats respectively, and the final concentration of the serum in both groups was adjusted to 5%. At three time points of 24 hours (T1), 48 hours (T2), and 72 hours (T3) of cell incubation, Western-blot technique was used to measure the protein expression levels of cytoskeletal proteins in PASMCs, including F-actin, α-tubulin, and Destrin. Meanwhile, the Transwell chamber (at the T1 time point) and the scratch assay (at the time points from T1 to T3) were used to detect the migration level of PASMCs. Results: Compared with Group C, the migration ability of PASMCs in the HPS group was significantly enhanced, while the protein expression of cytoskeletal proteins (F-actin, α-tubulin, and Destrin) showed a significant downward trend (P < 0.05). Conclusion: Under the stimulation of the serum of HPS rats, the expression of cytoskeletal proteins in PASMCs gradually decreases, which may play an important regulatory role in the abnormal migration of PASMCs related to HPS.