摘要
为明确黄芩苷对无乳链球菌(Streptococcus agalactiae,GBS)诱导的小鼠乳腺组织炎症反应的干预作用及机制,本研究将分娩后7 d~10 d的40只健康BALB/c泌乳小鼠随机分为阴性对照组、感染组、高中低剂量黄芩苷治疗组(20 mg/kg、40 mg/kg和80 mg/kg)。感染组和治疗组以GBS感染BALB/c小鼠,治疗组于感染24 h后每6 h经乳导管灌注相应剂量的黄芩苷一次,连续4次,最后一次给药6 h后摘取乳腺组织,HE染色后观察乳腺组织病理变化;利用RT-q PCR和间接ELISA检测各组小鼠乳腺组织中炎症因子及NLRP3炎症小体通路相关因子m RNA的相对转录水平及分泌水平;利用免疫组化试验检测各组小鼠乳腺组织中NLRP3蛋白的定位及含量的变化。组织病变观察可见,GBS感染可导致小鼠乳腺上皮细胞坏死、脱落,乳腺腺泡腔固有结构消失;与感染组相比,各治疗组不同浓度的黄芩苷均可减轻小鼠乳腺组织病理损伤。RT-q PCR检测结果显示,感染组小鼠乳腺组织中IL-1β、IL-6和TNF-αm RNA的相对转录水平显著或极显著高于阴性对照组小鼠(P<0.05或P<0.01);与感染组相比,各治疗组显著或极显著(P<0.05或P<0.01)下调IL-1β、IL-6、IL-10、TNF-α、Caspase-1、NLRP3和ASC基因m RNA的相对转录水平。ELISA检测结果显示,与感染组相比,各治疗组小鼠灌注不同浓度的黄芩苷可极显著(P<0.01)下调乳腺组织中IL-1β、Caspase-1和NLRP3蛋白的分泌水平,且免疫组化试验结果显示各治疗组小鼠乳腺组织中NLRP3蛋白分布减少。综上所述,40 mg/kg黄芩苷降低小鼠乳腺炎症反应效果最好,可更有效下调NLRP3炎症小体通路相关基因m RNA相对转录水平,抑制NLRP3炎症小体活化,降低乳腺组织炎症因子m RNA相对转录和分泌水平,缓解GBS诱导的小鼠乳腺组织炎性损伤。本研究为应用黄芩苷治疗GBS感染导致的奶牛乳腺炎提供了参考依据。
To clarify the intervention effect and mechanism of Baicalin on mammary tissue inflammation induced by Streptococcus agalactiae(GBS)in mice.In this study,40 healthy BALB/c lactating mice from 7 to 10 days after delivery were randomly divided into negative control group,infection group,high,medium and low dose Baicalin treatment group(20mg/kg,40mg/kg and 80mg/kg).The BALB/c mice from infection group and treatment group were infected with GBS.The treatment group mice were injected corresponding dose of Baicalin through the mammary duct once every 6 hours,for 4 consecutive times after 24 hours of GBS infection.6 hours after the last administration,the breast tissue was extracted and the pathological changes of breast tissue were observed by H.E.staining;ELISA and RT-qPCR were used to detect the transcription and secretion levels of inflammatory factors and NLRP3 inflammatory body pathway related factors in breast tissue of mice in each group.The localization and content of NLRP3 protein in breast tissue of mice in each group were detected by immunohistochemistry.Pathological observation showed that GBS infection caused necrosis and exfoliation of mouse mammary epithelial cells,and the disappearance of the inherent structure of mammary acinar cavity;Compared with infection group,different concentrations of Baicalin in each treatment group could reduce the pathological injury of mammary gland tissue.The results of RT-qPCR showed that the mRNA relative transcription levels of IL-1β,IL-6 and TNF-αin infection group mouse breast tissue was significantly higher than that in mice in the negative control group(P<0.05 or P<0.01).Compared with the infection group,Baicalin of different concentrations can reduce the pathological damage of mouse mammary tissue,significantly or extremely significantly(P<0.05 or P<0.01)down-regulate the mRNA transcription level of IL-1β,IL-6,IL-10,TNF-α,Caspase-1,NLRP3 and ASC genes.ELISA results showed that compared with infection group,different concentrations of scutellaria baicalae could s
作者
魏紫
冀禹彤
陈心瑜
吴昌旭
张金武
庞舒琪
韦英益
李军
彭昊
马春霞
胡庭俊
于美玲
WEI Zi;JI Yu-tong;CHEN Xin-yu;WU Chang-xu;ZHANG Jin-wu;PANG Shu-qi;WEI Ying-yi;LI Jun;PENG Hao;MA Chun-xia;HU Ting-jun;YU Mei-ling(College of Animal Science and Technology,Guangxi University,Nanning 530004,China;Guangxi Veterinary Research Institute,Nanning 530001,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2023年第9期951-958,共8页
Chinese Journal of Preventive Veterinary Medicine
基金
广西自然科学基金面上项目(2021GXNSFAA196004)
广西自然科学基金青年基金项目(2020GXNSFBA238018)。
关键词
无乳链球菌
小鼠乳腺炎症
NLRP3炎症小体
黄芩苷
Streptococcus agalactis(GBS)
mouse mammary gland inflammation
NLRP3 inflammasome
Baicalin
