Objective: To investigate whether glyburide prevents platelet-derived growth factor (PDGF) induced pulmonary artery smooth muscle cells(PASMCs) proliferation and migration via inhibiting nucleotide binding domain leucine-rich repeat-containing receptors protein 3(NLRP3) inflammasome activation. Methods: PASMCs were divided into 4 groups: control group, glyburide group, PDGF group and PDGF+ glyburide group. Cell proliferation and migration were assessed by MTT and Transwell respectively. The NLRP3 inflammasome activation was assessed by Western blot. Results: Compared with the control group, the protein expressions of NLRP3, caspase-1 and IL-1β in PASMCs were increased to (1.38±0.09, t=3.998, P<0.001), (1.32±0.1, t=3.268, P<0.01)and(1.43±0.19) (t=2.096, P<0.05) folds in the PDGF group. Glyburide had no effect on NLRP3, caspase-1 and IL-1β expression as compared with the control group, while the NLRP3, caspase-1 and IL-1β were decreased by(20.49±7.6)% (t=2.862, P<0.01), (32.94±3.44)% (t=4.154, P<0.001) and (24.67±5.29)% (t=2.335, P<0.05) in the PDGF+ glyburide group, respectively, as compared with the PDGF group. Compared with the control group, the PASMCs proliferation and migration in the PDGF group were significantly increased to (1.74±0.23, t=4.717, P<0.001) and (3.12±0.8, t=5.249, P<0.001) folds, respectively. Compared with the control group, glyburide had no effect on PASMCs proliferation and migration. In PDGF+ glyburide group, cell proliferation was reduced by (50.5±4.27)% (t=4.462, P<0.001) and cell migration count was lower than in the PDGF group (42.77±2.84)% (t=3.716, P<0.001). Conclusion: Glyburide could ameliorate PDGF-induced PASMCs proliferation and migration by inhibiting NLRP3 inflammasome activation.
目的： 探讨格列本脲是否通过抑制NLRP3炎性小体活化缓解肺动脉平滑肌细胞(PASMCs)增殖与迁移。 方法： 传代培养人PASMCs并分为4组：对照组、格列本脲组、血小板源性生长因子(PDGF)组和PDGF＋格列本脲组，实验组分别给予格列本脲干预和PDGF诱导PASMCs增殖和迁移。采用Western blot法检测各组细胞中NLRP3炎性小体的活化状况，噻唑蓝法检测细胞增殖率，Transwell法检测细胞迁移能力。 结果： PDGF组与对照组相比NLRP3表达升高至(1.38±0.09，t＝3.998, P<0.001)；半胱氨酸蛋白酶-1(caspase-1)表达增加到(1.32±0.10，t＝3.268, P<0.01)；IL-1β表达上调为(1.43±0.19，t＝2.096, P<0.05)。与对照组相比，格列本脲不影响NLRP3、caspase-1和IL-1β表达；PDGF＋格列本脲组与PDGF组相比NLRP3的表达降低(20.5±7.6)% (t＝2.862, P<0.01)，caspase-1表达减少(32.9±3.4)% (t＝4.154, P<0.001)，IL-1β表达下调(24.7±5.3)% (t＝2.335, P<0.05)。与对照组相比，PDGF组PASMCs增殖升高至(1.74±0.23，t＝4.717, P<0.001)，迁移增加到(3.12±0.8，t＝5.249, P<0.001)。与对照组相比，格列本脲不影响PASMCs的增殖与迁移；但可明显抑制PDGF诱导的PASMCs的增殖(50.5±4.3)% (t＝4.462, P<0.001)和迁移(42.77±2.84)% (t＝3.716, P<0.001)。 结论： 格列本脲可通过抑制NLRP3炎性小体的活化缓解PDGF诱导的PASMCs增殖与迁移。.
Keywords: Glyburide; Myocytes, smooth muscle; NLRP3; Platelet-derived growth factor.