干细胞水凝胶治疗奶牛子宫内膜炎的作用效果分析

Effect analysis of stem cell hydrogel in treatment of endometritis in dairy cows

摘要

基于间充质干细胞（MSCs）制剂在子宫内膜炎奶牛体内的治疗效果，探究干细胞制剂治疗奶牛子宫内膜炎的潜在机制。将奶牛分为健康组（n=5），子宫内膜炎症组（患子宫内膜炎但未进行任何处理，n=5），治疗组（患子宫内膜炎且宫内投入干细胞水凝胶制剂，n=5）和佐剂组（患子宫内膜炎且宫内投入不含干细胞的水凝胶制剂，n=5），利用牛用子宫内膜采样器采集子宫内膜组织，病理组织学和透射电镜观察奶牛的子宫上皮组织学变化，宫内投入制剂7 d后，利用牛用经直肠B超仪检测治疗效果，Western blot测量4组奶牛子宫内膜上皮组织中的高迁移率族蛋白B1（HMGB1）、半胱氨酸天冬氨酸特异性蛋白酶3（Caspase3）、消皮素E蛋白（GSDME）、白细胞介素-18（IL-18）和白细胞介素-1β（IL-1β）的蛋白含量。结果：HE染色和透射电镜显示，奶牛在患子宫内膜炎时，子宫内膜上皮细胞发生焦亡；B超检测发现，干细胞水凝胶能附着在子宫内膜上，经治疗后奶牛子宫腔内的图像由治疗前的高回声转变为无回声；Western blot结果显示，治疗组的GSDME和Caspase-3的蛋白浓度与佐剂组相比显著下降（P<0.05），与健康组相比无显著差异性（P>0.05），治疗组的Caspase-3蛋白表达量比炎症组显著下降（P<0.01），健康组和治疗组中的HMGB1蛋白与炎症因子IL-18、IL-1β无显著差异性（P>0.05），治疗组中HMGB1、IL-18和IL-1β的蛋白表达量比佐剂对照组、炎症组显著降低（P<0.01）。综上，干细胞水凝胶制剂通过抑制子宫上皮细胞焦亡进而显著降低了促炎因子IL-18和IL-1β的表达，在禁止抗生素滥用的背景下，为集约化牧场奶牛子宫内膜炎的治疗提供了新的方向。

Abstract

Based on the therapeutic effect of MSCs preparations in dairy cows with endometritis, this study was to explore the mechanism of the action of stem cell preparations in treating bovine endometritis. Twenty dairy cows were used and divided into a healthy group (n=5) and an endometritis group (endometritis without any treatment, n=5), a treatment group (endometritis with stem cell hydrogel preparation, n=5), and an adjuvant group (endometritis with stem cell free hydrogel preparation, n=5). Samples of the endometrial tissue were collected using a bovine endometrial sampler. Histological changes in the uterine epithelium of cows were observed through HE staining sections and transmission electron microscopy. Seven days after injection, the therapeutic effect was evaluated using bovine transrectal B-mode ultrasound. The protein contents of HMGB1, Caspase3, GSDME, IL-18, and IL-1β in the cow tissues were measured by Western blot in four groups. The results of the HE staining and transmission electron microscopy showed that, in the cows with endometritis, the endometrial epithelial cells underwent pyroptosis. The results of the B-mode ultrasound showed that the stem cell hydrogel attached to the endometrium, and the echo phenomenon in the cow uterus reduced or disappeared after treatment. The results of Western blot showed that the protein concentrations of GSDME and Caspase-3 in the treatment group were significantly reduced, compared with the adjuvant group (P<0.05). The protein concentrations of GSDME and Caspase-3 in the treatment group showed no significant difference, compared with the healthy group (P>0.05). The expression level of Caspase-3 protein in the treatment group was significantly lower than that in the inflammation group (P<0.01). There was no significant difference in HMGB1 protein and inflammatory factors IL-18 and IL-1β between the healthy group and the treatment group (P>0.05). The protein expression levels of HMGB1, IL-18, and IL-1β were significantly reduced in the treatment group, compared with the adjuvant control group and the inflammation group (P<0.01). To sum up, the stem cell hydrogel preparation played a good therapeutic role by inhibiting the scorch death of uterine epithelial cells. MSCs preparation significantly reduced the expression of pro-inflammatory factors IL-18 and IL-1β by inhibiting pyroptosis of the uterine epithelial cells. In the context of prohibiting antibiotic abuse, the present results suggest a new direction for treating endometritis in intensive dairy farms.

关键词

奶牛 / 子宫内膜炎 / 细胞焦亡 / 间充质干细胞 / 水凝胶

Key words

dairy cow / endometritis / cell pyroptosis / MSCs / hydrogel

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基金资助

黑龙江省“双一流”学科协同创新成果项目(LJGXCG2025-P36)