骨架蛋白在肝肺综合征大鼠血清诱导PASMCs迁移中的作用
Role of Cytoskeletal Proteins in Rat PASMCs Migration Induced by HPS Rat Serum
DOI: 10.12677/acm.2025.1541041, PDF,    科研立项经费支持
作者: 文 静*, 鲜俊杰, 陈 杨#:重庆市第七人民医院(重庆理工大学附属中心医院)麻醉科,重庆;陈 林:陆军军医大学第一附属医院麻醉科,重庆
关键词: 肝肺综合征肺动脉平滑肌细胞骨架蛋白Hepatopulmonary Syndrome Pulmonary Arterial Smooth Muscle Cells Cytoskeletal Proteins
摘要: 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 < 0.05)。结论:在HPS大鼠血清的刺激下,PASMCs中骨架蛋白表达逐步降低,这可能在HPS病理进程的PASMCs异常迁移中发挥重要调节作用。
Abstract: 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.
文章引用:文静, 鲜俊杰, 陈林, 陈杨. 骨架蛋白在肝肺综合征大鼠血清诱导PASMCs迁移中的作用[J]. 临床医学进展, 2025, 15(4): 1154-1159. https://doi.org/10.12677/acm.2025.1541041

参考文献

[1] Liu, L., Liu, N., Zhao, Z., Liu, J., Feng, Y., Jiang, H., et al. (2012) TNF‐α Neutralization Improves Experimental Hepatopulmonary Syndrome in Rats. Liver International, 32, 1018-1026. [Google Scholar] [CrossRef] [PubMed]
[2] Morrell, N.W., Adnot, S., Archer, S.L., Dupuis, J., Lloyd Jones, P., MacLean, M.R., et al. (2009) Cellular and Molecular Basis of Pulmonary Arterial Hypertension. Journal of the American College of Cardiology, 54, S20-S31. [Google Scholar] [CrossRef] [PubMed]
[3] 陈杨, 陈林, 杨勇, 等. 肝肺综合征大鼠血清诱导肺动脉平滑肌细胞增殖时Talin1和F-actin表达的变化[J]. 中华麻醉学杂志, 2019, 39(4): 471-474.
[4] 国斌, 易斌, 徐顺贵, 等. 肝肺综合征大鼠血清对肺微血管内皮细胞Akt表达的影响[J]. 中华麻醉学杂志, 2010, 30(1): 75-78.
[5] 王丽平, 李季声, 邓黎, 等. 高迁移率族蛋白B1对体外氧糖剥夺/复氧复糖后大鼠脊髓反应性星形胶质细胞不同亚型的作用[J]. 中国组织工程研究, 2023, 27(24): 3831-3837.
[6] Alishahedani, M.E., Yadav, M., McCann, K.J., Gough, P., Castillo, C.R., Matriz, J., et al. (2021) Therapeutic Candidates for Keloid Scars Identified by Qualitative Review of Scratch Assay Research for Wound Healing. PLOS ONE, 16, e0253669. [Google Scholar] [CrossRef] [PubMed]
[7] Melo-Silva, C.A., Gaio, E., Trevizoli, J.E., Souza, C.S., Gonçalves, A.S., Sousa, G.C.C., et al. (2011) Respiratory Mechanics and Lung Tissue Remodeling in a Hepatopulmonary Syndrome Rat Model. Respiratory Physiology & Neurobiology, 179, 326-333. [Google Scholar] [CrossRef] [PubMed]
[8] Soto, X., Li, J., Lea, R., Dubaissi, E., Papalopulu, N. and Amaya, E. (2013) Inositol Kinase and Its Product Accelerate Wound Healing by Modulating Calcium Levels, Rho Gtpases, and F-Actin Assembly. Proceedings of the National Academy of Sciences, 110, 11029-11034. [Google Scholar] [CrossRef] [PubMed]
[9] 应荣彪, 冯俊, 李建军, 等. αβ-微管蛋白在乳腺癌变不同阶段的表达及意义[J]. 中国癌症杂志, 2011, 21(8): 595-598.
[10] Kuure, S., Cebrian, C., Machingo, Q., Lu, B.C., Chi, X., Hyink, D., et al. (2010) Actin Depolymerizing Factors Cofilin1 and Destrin Are Required for Ureteric Bud Branching Morphogenesis. PLOS Genetics, 6, e1001176. [Google Scholar] [CrossRef] [PubMed]
[11] Zeng, J., Yi, B., Wang, Z., Ning, J., Wang, X. and Lu, K. (2013) Effect of Annexin A2 on Hepatopulmonary Syndrome Rat Serum-Induced Proliferation of Pulmonary Arterial Smooth Muscle Cells. Respiratory Physiology & Neurobiology, 185, 332-338. [Google Scholar] [CrossRef] [PubMed]