大鼠失血性休克后不同液体复苏对脾脏髓系抑制细胞数量变化影响的研究

发布时间：2018-06-10 03:17

本文选题：失血性休克 + 髓系抑制细胞　；参考：《蚌埠医学院》2013年硕士论文

【摘要】：研究背景及目的失血性休克是严重创伤及外科手术中常见的合并症，不仅表现在循环功能失衡，同时伴随着严重的免疫系统紊乱。研究表明,休克能增加宿主对外源性和内源性感染的易感性，促使感染率的增加，这与其引起的免疫抑制有关。近年来，一群髓系来源的细胞因在免疫系统中有负向调控的作用而受到关注，这群细胞首先在肿瘤患者体内发现，并逐渐统一命名为“髓系抑制细胞(myeloid—derivedsuppressorcells；MDSCs)”，参与肿瘤免疫逃逸。2007年，Delano等将这些细胞的研究由肿瘤学扩展到感染免疫领域。研究表明，MDSCs对休克后伴随的免疫反应产生重要影响。失血性休克后，炎症因子网络被触发，若不能及时终止其发生发展，常可导致免疫紊乱。目前，失血性休克后治疗的主要手段之一是液体复苏，但至今没有确凿证据表明哪种液体更好。本文研究的目的：建立一个稳定可靠的失血性休克模型，在此基础上初步探讨失血性休克不同液体复苏后脾脏中MDSCs数量变化，筛选出对失血性休克免疫抑制逆转作用较强的复苏液体并指导以后的干预治疗。对象及方法 1.72只SD大鼠，随机分成4组，，假手术组（Sham组）、高渗氯化钠复苏组（HRS组）、乳酸林格氏液复苏组（LRS组）、晶胶液复苏组（LCRS组），Sham组仅接受麻醉，置管及动脉监测，不予放血，其余组大鼠建立失血性休克模型。 2.失血性休克模型，参照Chaudry方法建立失血性休克模型，快速放血至已肝素化的储血瓶内，在10分钟内将平均动脉压降至40mmHg为目标，期间通过调节放血速度控制血压。 3.模型制作成功后，分别以不同的液体(高渗氯化钠液，乳酸林格氏液，晶胶液)进行复苏，复苏后12，24，48小时分别处死（颈椎脱位法处死大鼠）各组大鼠6只（包括假手术组），取脾脏组织。 4.将脾脏组织制作成细胞悬液，His48、CD11bc试剂双标后用流式细胞仪检测各组脾脏中MDSCs，计算MDSCs的比例变化。结果 1.参照Chaudry法成功建立失血性休克模型，达到平均动脉压40mmHg的要求。 2.假手术组在三个不同时间点脾脏中MDSCs数量变化在统计学上无明显差异。 3.相比于假手术组，失血性休克复苏后MDSCs数量均有不同程度的升高，差异明显，有统计学意义，其数量变化有先升后降的趋势。 4.三种不同的复苏液体，在三个不同时间点以高渗盐组复苏后MDSCs数量升高程度相对较小。12h时，与晶胶液相比，差异不明显，与乳酸林格氏液比，差异显著。但在24h,48h同另两组相比，具有统计学意义。结论 1.用Chaudry法建立的失血性休克模型，操作简便，易于复制，能反映失血性休克的发病过程和临床特征。 2.假手术组机体损伤较小，MDSCs变化无统计学差异，说明实验动物通过自我保护反应，维持机体免疫平衡。 3.失血性休克复苏后，机体的免疫功能早期受到不同程度的抑制，随时间变化逐渐恢复。 4.相对于乳酸林格氏液晶胶液，高渗氯化钠液复苏失血性休克，能有效抑制脾脏中MDSCs数量，减轻休克后早期的免疫抑制。
[Abstract]:Background and purpose of research
Hemorrhagic shock is a common complication in severe trauma and surgery, not only in the imbalance of circulatory function, but also with serious immune system disorders. Research shows that shock can increase the susceptibility to external and endogenous infection of the host and increase the infection rate, which is related to the immunosuppression caused by it. The cells derived from the medullary system are concerned about the role of negative regulation in the immune system. These cells are first found in the tumor patients and are gradually named "myeloid inhibitory cells (myeloid - derivedsuppressorcells; MDSCs)". They are involved in tumor immune escape for.2007 years, Delano and other cells are studied by oncology. It has been extended to the area of infection immunity. Studies have shown that MDSCs has an important effect on the immune response associated with shock. After hemorrhagic shock, the inflammatory factor network is triggered, and if it fails to terminate its development in time, it can often lead to immune disorders. At present, one of the main means of treatment after hemorrhagic shock is fluid resuscitation, but it has not been confirmed so far. The purpose of this study is to establish a stable and reliable model of hemorrhagic shock. On the basis of this, the changes in the number of MDSCs in the spleen after the resuscitation of hemorrhagic shock are preliminarily discussed, and the resuscitation fluid which has a strong reversal effect on the immunosuppression of hemorrhagic shock is screened and the intervention treatment will be guided in the future.
Object and method
1.72 SD rats were randomly divided into 4 groups, sham operation group (group Sham), hypertonic sodium chloride resuscitation group (group HRS), lactic acid Ringer's fluid resuscitation group (group LRS), crystal gel fluid resuscitation group (group LCRS), group Sham only receiving anesthesia, catheterization and arterial monitoring, no blood release, and the other rats were set up with hemorrhagic shock model.
2. model of hemorrhagic shock, a model of hemorrhagic shock was established by Chaudry method, and the average arterial pressure was reduced to 40mmHg within 10 minutes. The blood pressure was controlled by regulating the speed of bleeding.
After the 3. model was successfully made, the resuscitation was carried out with different liquid (hypertonic sodium chloride solution, lactic acid lactate solution, crystal glue), and 6 rats in each group (including sham operation group) were killed in 12,24,48 hours after resuscitation, and the spleen tissues were taken.
4. the spleen tissue was made into cell suspension. After His48 and CD11bc double labeling, the MDSCs in spleen of each group was detected by flow cytometry, and the proportion of MDSCs was calculated.
Result
1. the hemorrhagic shock model was successfully established according to the Chaudry method to achieve the 40mmHg requirement of mean arterial pressure.
2. in the sham operation group, there was no significant difference in the number of MDSCs in the spleen at three different time points.
3. compared with the sham operation group, the number of MDSCs after the resuscitation of hemorrhagic shock increased in varying degrees. The difference was significant, and the number of changes had the trend of rising and descending first.
4. three different resuscitation liquids, when the increase in the MDSCs number of the hypertonic salt group at three different time points was relatively small.12h, compared with the gel liquid, the difference was not obvious, and the difference was significant compared with the lactic acid Ringer's solution, but in 24h, 48h was statistically significant compared with the other two groups.
conclusion
1. the hemorrhagic shock model established by Chaudry method is simple and easy to replicate. It can reflect the pathogenesis and clinical characteristics of hemorrhagic shock.
2. in the sham operation group, the body damage was small, and the MDSCs change was not statistically significant, indicating that the experimental animals maintained their immune balance through self-protection response.
3. after resuscitation of hemorrhagic shock, the immune function of the body was inhibited in varying degrees, and gradually recovered with time.
4. the resuscitation of hemorrhagic shock with hypertonic sodium chloride solution can effectively inhibit the number of MDSCs in the spleen and reduce the early immunosuppression after shock.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R459.7

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