高浓度皮质醇对大肠杆菌诱导的山羊急性子宫内膜炎的作用研究

摘要

子宫内膜炎是一种主要影响产乳动物的产后疾病，通常由病原微生物感染引起。该疾病可引起产奶量下降，妊娠失败率增加，造成重大的经济损失。皮质醇是一种重要的炎症调节物质，其在炎症损伤中具有重要作用。在分娩过程中，母体血液中皮质醇水平升高。本实验重点观察，高浓度的皮质醇对山羊大肠杆菌性子宫内膜炎的调节作用机制。12只健康的山羊随机分为三组:对照组、大肠杆菌组和皮质醇治疗组。皮质醇治疗组，每天按1mg/kg的剂量每天静脉滴注皮质醇，3天后，子宫灌注大肠杆菌;大肠杆菌组每天静脉注射生理盐水，3天后，子宫灌注大肠杆菌;对照组每天静脉注射生理盐水，3天后，子宫灌注PBS。接种E.coli后0、12、24、96和168h，进行白细胞计数、体温、脉搏和呼吸的检测;接种E.coli后0、24、96和168h，进行血液皮质醇含量检测。收集子宫分泌物进行细胞学检查及细菌培养;采用HE染色法对子宫内膜的组织进行病理学观察;实时荧光定量PCR法检测子宫内膜组织中TLR4、IL-1β、IL-6、IL-8、TNF-α、TGF-β和VEGF。
　　大肠杆菌组和皮质醇治疗组的白细胞数量在12h时与对照组相比显著增加(p＜0.05)，感染24h后，恢复正常。大肠杆菌组和皮质醇治疗组在感染12h后体温升高。皮质醇治疗组血液皮质醇含量在每个时间点均高于空白组与大肠杆菌组。皮质醇治疗组感染大肠杆菌24h时，呼吸频率急剧上升，但在感染96h时恢复正常。三个组之间的心率没有明显变化。大肠杆菌组和皮质醇治疗组的子宫分泌物在接种24h时可见到大量的嗜中性粒细胞。大肠杆菌组及皮质醇治疗组子宫内膜组织中的白细胞数量在接种24h时与对照组相比升高，之后逐渐减少。感染12和24h时，大肠杆菌及皮质醇治疗组子宫分泌物内获得阳性细菌。大肠杆菌组和皮质醇治疗组子宫内膜组织中的TLR4、IL-1β和TNF-α的mRNA表达量在感染24h时与对照组相比没有显著差异，但皮质醇治疗组的IL-6和IL-8水平在感染后24h和96h时显著降低(p＜0.05)。在感染前后，大肠杆菌组和皮质醇治疗组子宫内膜组织中的TGF-β mRNA表达与对照组相比差异不显著。皮质醇治疗组的VEGF mRNA表达量在感染24h后与大肠杆菌组相比显著降低(p＜0.01)。

目录

摘要

Abstract

Table of contents

List of abbreviations

Introduction

1 Postpartum endometritis

1．1 Causes of postpartum endometritis

1．2 Symptoms of partum endometritis

1．3 Diagnosis of endometritis and subclinical endometritis

1．4 Treatment of endometritis in dairy cows

1．5 The postpartum uterus

1．6 Local immunity of the endometrium

1．7 Changes in the uterus during endometritis

2 Cortisol

2．1 Glucocorticoids

2．2 Mechanism of action

2．3 Glucocorticoids and inflammation

2．4 Anti-Inflammatory Effects of Glucocorticoids

2．5 Pro-Inflammatory Effects of Glucocorticoids

2．6 Glucocorticoids and recovery

2．7 Glucocorticoids during pregnancy

References

Experiment 1：The effect of cortisol on the physical and blood parameters of goats with endometritis induced by E．coli

1 Materials

1．1 Animals

1．2 Instruments

1．3 Reagents

1．4 Prepared reagents

2 Experimental method

2．1 Preparation of bacteria

2．2 Preparation of animals and groups

2．4 Clinical inspection

2．5 Uterine discharge cytology

3 Experimental results

3．1 Clinical inspection

3．2 Uterine cytology examination results

4 Discussion

References

Experiment 2：The effect of cortisol on the pathological tissue structure of goats with endometritis induced by E．coli

1．3 Reagents

1．4 Prepared reagents

2 Experimental methods

2．2 Preparation of paraffin sections

3 Results

4 Discussion

References

Experiment 3：The effect of cortisol on mRNA expression of TLR4 and other inflammatory cytokines in endometrial tissue of the goat with endometritis induced by E．coli

1 Materials

1．3 Reagents

1．4 Prepared reagents

2 Methods

2．1 Collection and preparation of sample

2．2 Real-time fluorescence quantitative PCR

3 Results

3．2 The effect of elevated cortisol levels on mRNA expression of inflammatory factors in the endometrial tissue of E．coli induced endometritis in goats

3．3 The effect of elevated cortisol levels on mRNA expression of recovery related genes in the endometrial tissue of E．coli induced endometritis in goats

4 Discussion

References

Conclusion

Acknowledgment

声明