Changes in expression of Talin1 and F-actin during serum-induced proliferation of pulmonary artery smooth muscle cells of rats with hepatopulmonary syndrome

Abstract

Objective To evaluate the changes in the expression of Talin1 and F-actin during serum-induced proliferation of pulmonary artery smooth muscle cells (PASMCs) of rats with hepatopulmonary syndrome (HPS). Methods Twenty healthy male Sprague-Dawley rats, weighing 220-250 g, were used for producing HPS by chronic ligation of the common bile duct.Blood samples from the abdominal aorta were collected to prepare serum.Primarily cultured PASMCs obtained from rats were seed in 6- or 96-well plates and divided into 2 groups using a random number table method: control group (group C) and HPS group (group HPS), with 24 wells in each group (for 6-well plates) or with 30 wells in each group (for 96-well plates). In C and HPS groups, normal rat serum or HPS rat serum were added, respectively, with the final concentration of 5%.At 24, 48 and 72 h of incubation, the expression of Talin1, F-actin and G-actin was determined by Western blot, the F-actin/G-actin ratio was calculated, and the proliferation of PASMCs was measured by 3H-TdR incorporation and CCK-8 assays. Results Compared with group C, the proliferation of PASMCs was significantly enhanced, the expression of Talin1 was up-regulated, and the F-actin/G-actin ratio was increased in group HPS (P<0.05). The proliferation of PASMCs was gradually enhanced, the expression of Talin1 was gradually up-regulated, and the F-actin/G-actin ratio was gradually increased with the prolonged incubation time in group HPS (P<0.05). Conclusion The mechanism by which the HPS rat serum induces proliferation of PASMCs may be related to up-regulating the expression of Talin1 and F-actin. Key words: Hepatopulmonary syndrome; Myocytes, smooth muscle; Pulmonary artery; Cell proliferation; Talin; Actins