转染Cox7a2重组质粒对大鼠支持细胞分泌功能和ERK磷酸化水平的影响

Analysis of cell secretion function and the phosphorylation level of extracellular signal-regulated kinase in rat sertoli cell transfected with Cox7a2 recombinant plasmid

目的：观察大鼠睾丸支持细胞转染细胞色素C氧化酶7a2（Cox7a2）重组质粒后对支持细胞分泌功能及细胞外信号调节激酶（ERK）磷酸化水平的影响。方法培养原代大鼠睾丸支持细胞，将细胞分为空白对照组、空载体组、重组质粒组。将pEGFP-C1-Cox7a2重组质粒转染支持细胞24h后，收集细胞上清液，采用酶联免疫法，测定转铁蛋白（TF）、白细胞介素-1（interleukin-1, IL-1）β、白细胞介素-6（IL-6）含量。Western blot检测ERK的磷酸化水平。结果重组质粒组TF为（14.11±0.45）pmol/mL，显著低于空白对照组（24.3±0.64）pmol/mL和空载体组（25.16±0.42）pmol/mL（P＜0.01）。重组质粒组IL-1β、IL-6水平分别为（157.14±12.69）pg/mL、（0.79±0.04）pg/mL，显著高于空白对照组（33.05±1.92）pg/mL、（0.25±0.01）pg/mL和空载体组（40.46±6.69）pg/mL、（0.37±0.03）pg/mL，差异均具统计学意义（P＜0.01）。与空白对照组和空载体组相比，重组质粒组ERK1/2磷酸化水平明显增高（P＜0.01）。结论转染Cox7a2重组质粒后损害了大鼠睾丸支持细胞的分泌功能，提高ERK1/2磷酸化水平。

Objective To investigate cell secretion function and the phosphorylation level of extracellular signal-regulated kinase in rat sertoli cell transfected with Cox7a2 recombinant plasmid. Methods Rat SCs were cultured and divided into three groups such as the control group, the mock group and the recombinant plasmid group. After 24h culture, the culture supernatants of the transfected cells were collected and the levels of transferrin （TF） , interleukin（IL）-1β and IL-6 in culture medium were measured by ELISA. The level of ERK phosphorylation in the transfected cells was detected by Western Blot. Results The level of TF was significantly decreased in the recombinant plasmid group as compared with that of the control group and the mock group （P〈0.05） .The levels of IL-1βand IL-6 （157.14±12.69 pg/mL and 0.79±0.04 pg/mL） in the recombinant plasmid group were significantly higher than those of the control group and the mock group（33.05±1.92 pg/mL, 0.25±0.01pg/mL;40.46±6.69pg/mL, 0.37±0.03pg/mL, P〈0.01）, and the level of ERK phosphorylation was also increased significantly（P〈0.01）. Conclusion The cell secretion function was impaired in rat SCs transfected with Cox7a2 recombinant plasmid but the phosphorylation level of ERK was increased.