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目的:探讨瘦素对大鼠气道平滑肌细胞(ASMCs)增殖的影响及其可能的作用机制。方法:体外培养大鼠的ASMCs,分别用RT-PCR和Western blot测定ASMCs中瘦素受体mRNA和该细胞上瘦素受体蛋白的表达。不同浓度的瘦素(0～100μg/L)干预培养的ASMCs不同时间(1～72 h)后,以CCK-8法测定ASMCs的增殖情况。不同浓度的瘦素作用于ASMCs 48 h后,Western blot测定磷酸化细胞外调节蛋白激酶(p-ERK)和磷脂酰肌醇3激酶(PI-3K)的表达。结果:不同浓度的瘦素作用不同时间后,均可促进大鼠ASMCs增殖,并呈浓度依赖性(r=0.837,P<0.01)和时间依赖性(r=0.874,P<0.01)。Western blot的结果显示,不同浓度的瘦素干预后,大鼠ASMCs中p-ERK、PI-3K蛋白的表达较对照组显著增加(P<0.05),并与瘦素的浓度呈正相关(前者r=0.793,P<0.01,后者r=0.746,P<0.01)。结论:大鼠ASMCs表面有瘦素受体表达。瘦素可促进体外培养的大鼠ASMCs增殖,其机制可能与激活p-ERK和PI-3K有关。
Objective: To investigate the effect of leptin on the proliferation of rat airway smooth muscle cells (ASMCs) and its possible mechanism. Methods: Rat ASMCs were cultured in vitro. The expression of leptin receptor mRNA and leptin receptor protein in ASMCs were detected by RT-PCR and Western blot respectively. After different concentrations of leptin (0 ~ 100μg / L) interfered with cultured ASMCs for different time (1 ~ 72 h), the proliferation of ASMCs was determined by CCK-8 assay. After different concentrations of leptin were treated with ASMCs for 48 h, the expressions of phosphorylated extracellular regulated protein kinase (p-ERK) and phosphatidylinositol 3-kinase (PI-3K) were determined by Western blot. Results: Different concentrations of leptin promoted proliferation of rat ASMCs in a concentration-dependent manner (r = 0.837, P <0.01) and time-dependent manner (r = 0.874, P <0.01) at different time points. The results of Western blot showed that the expressions of p-ERK and PI-3K in ASMCs were significantly increased (P <0.05), and positively correlated with the concentrations of leptin after treated with different concentrations of leptin (the former r = 0.793, P <0.01, the latter r = 0.746, P <0.01). Conclusion: The expression of leptin receptor on the surface of rat ASMCs. Leptin can promote the proliferation of rat ASMCs cultured in vitro, the mechanism may be related to the activation of p-ERK and PI-3K.