镉处理对小鼠睾丸间质细胞PI3K/AKT信号通路的影响

目的 探讨镉对小鼠睾丸间质细胞3-磷酸肌醇激酶/蛋白激酶B(PI3K/AKT)信号通路的影响.方法 TM3细胞经20μmol/L CdCl2处理,分别在加入CdCl24 h和8h后收集细胞;对照组给予相应体积的PBS,8h后收集细胞.利用实时定量RT-PCR法测定炎症相关细胞因子白细胞介素(IL)-10、肿瘤坏死因子(TNF)-α、IL-6、单核细胞趋化蛋白(MCP)-1、巨噬细胞炎性蛋白(MIP)-2、MIP-1和环氧合酶(COX)-2 mRNA的表达水平以及用Western blot法检测细胞COX-2、AKT和p-AKT蛋白的表达水平.结果 与对照组相比,镉能够显著诱导TNF-α和IL-6 mRNA的表达(P＜0.01);镉处理4h组MCP-1 mRNA表达水平明显高于对照组(P＜0.01),而镉处理8h组MCP-1 mRNA表达水平与对照组相比差异无统计学意义;镉对TM3细胞MIP-1和MIP-2mRNA表达差异无统计学意义;镉能够明显诱导TM3细胞COX-2蛋白和mRNA的表达(P＜0.01).此外,镉处理组p-AKT蛋白表达水平也明显高于对照组(P＜0.01).结论 镉可能通过激活TM3细胞中PI3 K/AKT信号通路进而选择性调节部分炎症相关细胞因子的表达.

Effect of cadmium on PI3K/AKT signal pathway in mouse leydig cells

Objective To study the effects of cadmium on phosphatidylinositol 3-kinase/protein kinase B (PI3K/ AKT) signal pathway in mouse leydig cells.Methods TM3 cells (mouse leydig cell line) were incubated with 20 μmol/L CdCl2 and collected cells at 4 h and 8 h after cadmium exposure.The control group was incubated with corresponding volume PBS and collected cells after 8 h.The mRNA expression levels of interleukin(IL)-10,tumor necrosis factor (TNF)-α,IL-6,monocyte chemotactic protein (MCP)-1,macrophage inflammatory protein (MIP)-2,MIP-1 and cyclooxygenase(COX)-2 were detected by RT-PCR;the protein expression levels of COX-2,AKT and p-AKT were measured by Western blot.Results Compared with the control group,cadmium significantly increased the mRNA expression levels of IL-6 and TNF-α (P ＜ 0.01);the mRNA expression level of MCP-1 was also increased at 4 h group after cadmium exposure (P ＜0.01),however,cadmium did not affects the mRNA expression level of MCP-1 at 8 h group after cadmium exposure.No significant differences on the mRNA expression levels of MIP-1 and MIP-2 were observed between the control group and cadmium groups.Cadmium markedly upregulated the mRNA and protein levels of COX-2 in TM3 cells (P ＜ 0.01).In addition,compared with the control group,the protein expression of p-AKT also was significantly increased in cadmium groups (P ＜ 0.01).Conclusion These results suggest that cadmium could activate PI3K/AKT signal pathway which might partially contribute to cadmium-induced the secretion of some inflammatory cytokines in mouse testicular leydig cells.