啶虫脒对小鼠睾丸间质细胞活力、凋亡的影响及其机制探讨

Effects and mechanism of acetamiprid on viability and apoptosis of Leydig cells

目的 探讨啶虫脒(ACE)对小鼠睾丸间质细胞TM3的活力、凋亡的影响及其作用机制,为ACE生殖毒性及机制研究提供线索。方法 以0、2、4、6、8 mmol/L ACE分别加入小鼠睾丸间质细胞TM3细胞(A0、A1、A2、A3、A4组)中作用24 h, CCK8法检测细胞活力,qPCR检测Caspase 3、Bcl-2、Bax及磷脂酰肌醇-3-激酶(PI3K)、蛋白激酶B(AKT)、叉头框蛋白1(FOXO1)mRNA。结果 与A0组比较,A2、A3、A4组细胞存活率均降低,差异有统计学意义(P均<0.01)。与A0组比较,A1、A2组Caspase 3 mRNA升高,A3、A4组Bcl-2 mRNA、Bax mRNA、Bcl-2/Bax升高(P均<0.05)。与A0组比较,A3、A4组PI3K mRNA升高(t分别为7.759、3.375,P均<0.05),A2、A3、A4组AKT mRNA升高(t分别为6.158、2.972、3.399,P均<0.05),A1、A2、A3、A4组FOXO1 mRNA升高(P均<0.05)。结论 ACE可导致TM3细胞活力降低,剂量越高作用越强;低剂量ACE可促进TM3细胞凋亡,高剂量ACE可抑制细胞凋亡;高剂量ACE可能通过调控PI3K/AKT/FOXO1信号通路发挥抗凋亡作用。

Objective To explore the effects of acetamiprid(ACE) on cell viability and apoptosis of mouse Leydig cells TM3 and its mechanism, so as to provide clues for the mechanism study of ACE reproductive toxicity. Methods TM3 cells were treated with 0,2,4,6 and 8 mmol/L ACE(A0,A1,A2,A3 and A4 groups) for 24 h respectively.Cell viability was detected by CCK8 assay.The mRNA levels of Caspase 3,Bcl-2,Bax, PI3K,AKT and FOXO1 were determined by qPCR. Results Compared with A0 group, the cell viability in A2,A3 and A4 groups was significantly decreased(P<0.01).qPCR results showed that, compared with A0 group, Caspase 3 mRNA in A1 and A2 groups, Bcl-2 mRNA,Bax mRNA and Bcl-2/Bax in A3 and A4 groups were significantly increased(all P<0.05).Compared with A0 group, PI3K mRNA in A3 and A4 groups increased(t=7.759 and 3.375,respectively, all P<0.05),AKT mRNA in A2,A3 and A4 groups increased(t=6.158,2.972 and 3.399,respectively, all P<0.05),and FOXO1 mRNA increased in A1,A2,A3 and A4 groups(all P<0.05). Conclusion ACE can reduce the viability of TM3 cells, and the higher the dose, the stronger the effect.Low-dose ACE can promote TM3 cell apoptosis, and high-dose ACE can inhibit cell apoptosis.ACE may play an anti-apoptotic effect by regulating PI3K/AKT/FOXO1 signaling pathway.