Department of Respiratory Diseases, Henan Province People's Hospital, Zhengzhou 450003, China.
Chin J Integr Med. 2010 Aug;16(4):331-6. doi: 10.1007/s11655-010-0505-1. Epub 2010 Aug 10.
To investigate the relationship between the proliferation of sensitized human airway smooth muscle cells (HASMCs) and the expression of extracellular signal regulated kinase (ERK) and the effect of Shenmai Injection (SMI) on HASMCs.
The HASMCs cultured in vitro were divided into three groups: (1) control group; (2) sensitized group: containing 10% asthmatic serum; (3) SMI group: further divided into three different concentration subgroups interferred with 10 microL/mL, 50 microL/mL, and 100 microL/mL SMI, respectively. The proliferation of HASMCs was detected using MTT method, the expression of proliferating cell nucleus antigen (PCNA) in HASMCs was detected using immunocytochemical staining, and the expression of phosphoration-ERK1/2 (p-ERK1/2) protein was detected using Western-blot.
After passive sensitization,: the optical density value (A A(490) value) of HASMCs was significantly increased from 0.366+/-0.086 to 0.839+/- 0.168 (P<0.05). In addition, the expression of PCNA was significantly increased from 28.7%+/-5.9% in the control group to 69.8%+/-7.5% in the sensitized group (P<0.05). At the same time, the expression of p-ERK1/2 in passively sensitized HASMCs was significantly increased compared with the control group (all P<0.05). After application of 10 microL/mL, 50 microL/mL, and 100 microL/mL SMI to the cultured media of passively sensitized group, the A(570) value was significantly decreased from 0.839+/-0.168 to 0.612+/-0.100, 0.412+/-0.092, and 0.339+/-0.077, respectively (P<0.05). Moreover, the expression of PCNA was significantly decreased from 69.8%+/-7.5% to 57.8%+/-6.2%, 40.7%+/-5.4%, and 26.1%+/-5.2%, respectively. At the same time, the expression of p-ERK1/2 in each SMI group was significantly decreased compared with the sensitized group (all P<0.05).
ERK signal transduction pathway may be involved in the airway remodeling in asthma. The expression of ERK can be inhibited by SMI in a dose-dependent manner, thus preventing the proliferation of HASMCs.
研究致敏人气道平滑肌细胞（HASMCs）增殖与细胞外信号调节激酶（ERK）表达的关系，以及参麦注射液（SMI）对 HASMCs 的作用。
体外培养 HASMCs，分为三组：（1）对照组；（2）致敏组：含 10%哮喘患者血清；（3）SMI 组：进一步分为三组，分别用 10μL/mL、50μL/mL、100μL/mL 的 SMI 进行干预。采用 MTT 法检测 HASMCs 的增殖，免疫细胞化学染色法检测增殖细胞核抗原（PCNA）在 HASMCs 中的表达，Western-blot 法检测磷酸化-ERK1/2（p-ERK1/2）蛋白的表达。
被动致敏后，HASMCs 的吸光度值（A A（490）值）从 0.366+/-0.086 显著增加到 0.839+/-0.168（P<0.05）。此外，与对照组相比，致敏组 PCNA 的表达从 28.7%+/-5.9%显著增加到 69.8%+/-7.5%（P<0.05）。同时，与对照组相比，被动致敏 HASMCs 中 p-ERK1/2 的表达显著增加（均 P<0.05）。在将 10μL/mL、50μL/mL 和 100μL/mL 的 SMI 应用于被动致敏组的培养介质后，A（570）值分别从 0.839+/-0.168 显著降低至 0.612+/-0.100、0.412+/-0.092 和 0.339+/-0.077（P<0.05）。此外，PCNA 的表达从 69.8%+/-7.5%显著降低至 57.8%+/-6.2%、40.7%+/-5.4%和 26.1%+/-5.2%，而各 SMI 组的 p-ERK1/2 表达均显著低于致敏组（均 P<0.05）。
ERK 信号转导通路可能参与哮喘气道重塑。SMI 可呈剂量依赖性抑制 ERK 的表达，从而阻止 HASMCs 的增殖。
