奶牛子宫内膜上皮细胞氧化应激模型的建立及奶牛脂肪间充质干细胞对其迁移的影响

doi:10.16431/j.cnki.1671-7236.2022.07.008

Abstract

Abstract: 【Objective】 The aim of this study was to explore the effect of bovine adipose-derived mesenchymal stem cells (bAD-MSCs) on the migration of bovine endometrial epithelial cells under oxidative stress.【Method】 Bovine endometrial epithelial cells were treated with 0,5,25,50,100,200 μmol/L H₂O₂ for 2,4,6,8 and 12 h,activity was detected by MTT assay and intracellular reactive oxygen species (ROS) level was detected by flow cytometry,in order to screen the best optimum condition of oxidative stress model of bovine endometrial epithelial cells.Six groups including control group,H₂O₂ group,bAD-MSCs co-culture group (1∶0.5),bAD-MSCs co-culture group (1∶1),MACT co-culture group (1∶0.5)and MACT co-culture group (1∶1) were established in the cell scratch test,and the migration ability of bovine endometrial epithelial cells was analyzed,the expression levels of extracellular regulated protein kinases (Erk) and phosphorylation extracellular regulated protein kinases (pErk) protein were detected by Western blotting.【Result】 The results of MTT assay showed that the cell survival rate of 4-12 h was 50%-80% when H₂O₂ concentration was 50 μmol/L,which was suitable for the establishment of oxidative stress model.Flow cytometry assay results showed that when bovine endometrial epithelial cells were stimulated with 50 μmol/L H₂O₂ for 2 h,the ROS level was significantly higher than that in control group and 4,6,8,12 h groups (P<0.05).The results of scratch test indicated that compared with control group,the migration ability of bovine endometrial epithelial cells in H₂O₂ group was significantly reduced (P＜0.05);While compared with H₂O₂ group,the migration ability of the bovine endometrial epithelial cells in bAD-MSCs co-culture groups was significantly increased (P＜0.05),and the cell migration ability in MACT co-culture groups was significantly decreased (P＜0.05).The results of Western blotting showed that compared with control group,the expression of pErk protein in bovine endometrial epithelial cells in H₂O₂ group was significantly decreased (P＜0.05);While compared with H₂O₂ group,the expression of pErk protein in bovine endometrial epithelial cells in bAD-MSCs co-culture groups was significantly increased (P＜0.05),and the expression of pErk protein in MACT co-culture groups was significantly decreased (P＜0.05).【Conclusion】 50 μmol/L H₂O₂ treatment for 2 h could successfully construct the oxidative stress model of bovine endometrial epithelial cells,and bAD-MSCs could promote the migration of bovine endometrial epithelial cells under oxidative stress and participate in regulating the expression of Erk protein.

Key words: bovine adipose-derived mesenchymal stem cells; oxidative stress; bovine endometrial epithelial cells; cell migration

CLC Number:

S857.2⁺1

LU Wengeng, SI Linqing, TIAN Chunyu, YUAN Siqi, XIE Hexin, ZHOU Jiwei, JIN Jidong, HAN Yinghao. Establishment of Oxidative Stress Model of Bovine Endometrial Epithelial Cells and the Effects of Bovine Adipose-derived Mesenchymal Stem Cells on Cell Migration[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2506-2514.

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Establishment of Oxidative Stress Model of Bovine Endometrial Epithelial Cells and the Effects of Bovine Adipose-derived Mesenchymal Stem Cells on Cell Migration

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