Feng Xiaowei, Li Yan, Wen Xiangfu, Feng Mingque, Jin Tianxiong, Shahid Muhammad, Sun Yan, Liu Jiawei, Liu Bei, Cheng Jia, Liu Mingchao
College of Veterinary Medicine, Hebei Agricultural University, Baoding, China.
Key Laboratory of Healthy Breeding in Dairy Cattle (Co-construction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Hebei Agricultural University, Baoding, China.
Anim Biosci. 2025 Sep;38(9):1996-2007. doi: 10.5713/ab.25.0031. Epub 2025 Jun 4.
The objective of this study was to evaluate the function of the reactive oxygen species (ROS)-mitochondrial pathway in attenuating Escherichia coli (E. coli) induced apoptosis in bovine endometrial epithelial cells (BENDs) by Lactobacillus rhamnosus (L. rhamnosus) GR-1.
The BENDs were exposed to preincubation with and without L. rhamnosus GR-1 for 3 hours (h) and they were later subject to E. coli for 6 h. The release of lactate dehydrogenase (LDH), the expression of oxidative factors, adhesion and invasion of E. coli, the expression of mitochondrial membrane potential (MMP), apoptotic rate and apoptosis-associated protein expression were observed. Then, ROS expression, MMP level and cell rate apoptosis rate were further detected after the intervention of antioxidant n-acetyl-l-cysteine (NAC).
L. rhamnosus GR-1 was capable of obviously alleviating the content of LDH, ROS expression, E. coli adhesion and invasion, the apoptotic rate and MDA concentrations in BENDs induced by E. coli (p<0.01). In addition, L. rhamnosus GR-1 could notably promote the levels of antioxidant factors (SOD, GSH, T-AOC) (p<0.01), inhibit the depolarization of MMP (p<0.01), and levels of apoptosis-related factors (Bax, Caspase-3 and Cyt-c), and promote anti-apoptosis factor (Bcl-2) (p<0.01). Moreover, after the intervention of antioxidant NAC, ROS expression, MMP depolarization, and apoptotic rate induced by E. coli in BENDs were decreased (p<0.01).
In E. coli-induced apoptosis in BENDs, therapeutic strategies aimed at down-regulating ROS and MDA and up-regulating antioxidant factors may maintain mitochondrial function and provide theoretical support for the use of probiotics in bovine endometritis.
本研究旨在评估活性氧（ROS）-线粒体途径在鼠李糖乳杆菌（L. rhamnosus）GR-1减轻大肠杆菌（E. coli）诱导的牛子宫内膜上皮细胞（BENDs）凋亡中的作用。
将BENDs分别在有或无鼠李糖乳杆菌GR-1的条件下预孵育3小时（h），随后使其暴露于大肠杆菌中6小时。观察乳酸脱氢酶（LDH）的释放、氧化因子的表达、大肠杆菌的黏附与侵袭、线粒体膜电位（MMP）的表达、凋亡率及凋亡相关蛋白的表达。然后，在抗氧化剂N-乙酰-L-半胱氨酸（NAC）干预后，进一步检测ROS表达、MMP水平及细胞凋亡率。
鼠李糖乳杆菌GR-1能够显著减轻大肠杆菌诱导的BENDs中LDH的含量、ROS表达、大肠杆菌的黏附与侵袭、凋亡率及丙二醛（MDA）浓度（p<0.01）。此外，鼠李糖乳杆菌GR-1可显著提高抗氧化因子（超氧化物歧化酶、谷胱甘肽、总抗氧化能力）的水平（p<0.01），抑制MMP的去极化（p<0.01），以及凋亡相关因子（Bax、半胱天冬酶-3和细胞色素c）的水平，并促进抗凋亡因子（Bcl-2）的表达（p<0.01）。此外，在抗氧化剂NAC干预后，大肠杆菌诱导的BENDs中的ROS表达、MMP去极化及凋亡率均降低（p<0.01）。
在大肠杆菌诱导的BENDs凋亡中，旨在下调ROS和MDA并上调抗氧化因子的治疗策略可能维持线粒体功能，并为益生菌在牛子宫内膜炎中的应用提供理论支持。
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