三磷酸肌醇受体各亚型在大鼠气道平滑肌细胞中的表达以及在慢性哮喘中的作用

目的检测正常大鼠气道平滑肌细胞中三磷酸肌醇受体(Inositol 1,4,5 triphosphate receptors, IP3R)各亚型的表达以及在慢性哮喘中的作用.方法采用胶原酶消化法培养大鼠气道平滑肌细胞,RT-PCR方法测定IP3R亚型的表达,并将纯化的PCR产物与pGEM-T载体连接后进行DNA序列分析;卵蛋白致敏并激发大鼠制备慢性哮喘模型,RT-PCR测定各受体mRNA表达水平的变化.结果大鼠气道平滑肌细胞中IP3R的3种亚型共表达,测序结果与GenBank中登录的序列完全一致;制备的慢性哮喘模型中,病理切片显示平滑肌层及粘膜层明显增厚,IP3R1的表达较对照组有明显的上调P＜0.01.结论大鼠气道平滑肌细胞中IP3R 3种亚型共表达,提示存在着复杂的细胞内钙调节机制;并且IP3R1在慢性哮喘的发生过程中可能起了一定的作用.

Expression of inositol 1,4,5-triphosphate receptor in airway smooth muscle cells of rats and its roles in chronic asthma formation

Objective To detect the expression of inositol 1,4,5-triphosphate receptor (IP 3R) subtypes in normal rat airway smooth muscle cells (ASMCs) and changes during chronic asthma formation. Methods ASMCs were cultured by collagen enzyme digestion method. The expressions of subtypes of IP 3R were detected by RT-PCR and the purified PCR products were linked with pGEM-T vector for DNA sequencing. Chronic asthma model was established with egg albumin. The changes of IP 3Rs were detected by RT-PCR method. Results All subtypes of IP 3R were expressed in airway smooth muscle cells of normal rats. The expression of IP 3R1 in asthma groups increased obviously as compared with that in the control group (P<0.01). Conclusion Co-expression of three subtypes of IP 3R in ASMCs of normal rats indicate that there is complicated intercellular Ca 2 regulation mechanism in chronic asthma.