Effects of microcystin-LR on proliferation and apoptosis of bovine endometrial epithelial cells in vitro

Treating bovine endometrial epithelial cells with Microcystin-LR in vitro explore its effects on cell viability and the mechanism of molecular regulation, in order to provide theoretical and practical basis for improving the efficiency of bovine reproduction. CCK-8 was used to detect cell survival rate after treatment; flow cytometry was used to detect cell cycle changes and cell apoptosis rate; Annexin V-FITC kit was used to detect cell apoptosis; real-time fluorescence quantitative detection of Pi3k, Akt, mTOR, Cyt-c, Caspase-3, Caspase-9 and p53 gene mRNA expression levels. Western blot was used to detect the protein expression of Pi3k, Akt, Caspase-3 and Caspase-9. The cell survival rate under different concentrations and time treatments showed two opposite results: increase and decrease. According to the CCK-8 test results, two treatment options of 12 h, 100 μg · L^-1and 24 h,160 μg·L^-1were selected for follow-up experiments. At 12 h, 100 μg·L^-1treatment conditions, the mRNA expression of Pi3k, Akt, mTOR were increased（P＜0.01）, and the mRNA expression of Cyt-c, Caspase-3and Caspase-9 were genes decreased（P＜0.05）. The expression of Pi3k and Akt protein were increased（P＜0.05）, and the expression of Caspase-3 and Caspase-9 protein were decreased（P＜0.05）. The number of cells in the G0/G1 phase decreased（P＜0.01）, and the number of cells in the G2/M and S phase was increased（P＜0.01）; the mRNA expression of p53 gene was increased（P＜0.05）. In 24 h, 160μg·L^-1treatment conditions, Caspase-3, Caspase-9, Bax/Bcl2 gene mRNA were expression increased（P＜0.05）, Caspase-3 and Caspase-9 protein expression were increased（P＜0.05）. Cells were treated with100 μg · L^-1MC-LR for 12 h, cells promoted cell proliferation by enhancing Pi3k-Akt-mTOR signaling pathway and inhibiting mitochondrial apoptosis pathway, reducing the number of cells in G1 phase, and increasing cells in G2/M and S phases. After treating the cells with 160 μg · L^-1MC-LR for24 h, the cells undergo apoptosis by activating the mitochondrial apoptosis pathway.