本文選題：失血性休克 + 髓系抑制細(xì)胞　； 參考：《蚌埠醫(yī)學(xué)院》2013年碩士論文

【摘要】：研究背景及目的 失血性休克是嚴(yán)重創(chuàng)傷及外科手術(shù)中常見(jiàn)的合并癥，不僅表現(xiàn)在循環(huán)功能失衡，同時(shí)伴隨著嚴(yán)重的免疫系統(tǒng)紊亂。研究表明,休克能增加宿主對(duì)外源性和內(nèi)源性感染的易感性，促使感染率的增加，這與其引起的免疫抑制有關(guān)。近年來(lái)，一群髓系來(lái)源的細(xì)胞因在免疫系統(tǒng)中有負(fù)向調(diào)控的作用而受到關(guān)注，這群細(xì)胞首先在腫瘤患者體內(nèi)發(fā)現(xiàn)，并逐漸統(tǒng)一命名為“髓系抑制細(xì)胞(myeloid—derivedsuppressorcells；MDSCs)”，參與腫瘤免疫逃逸。2007年，Delano等將這些細(xì)胞的研究由腫瘤學(xué)擴(kuò)展到感染免疫領(lǐng)域。研究表明，MDSCs對(duì)休克后伴隨的免疫反應(yīng)產(chǎn)生重要影響。失血性休克后，炎癥因子網(wǎng)絡(luò)被觸發(fā)，若不能及時(shí)終止其發(fā)生發(fā)展，�？蓪�(dǎo)致免疫紊亂。目前，失血性休克后治療的主要手段之一是液體復(fù)蘇，但至今沒(méi)有確鑿證據(jù)表明哪種液體更好。本文研究的目的：建立一個(gè)穩(wěn)定可靠的失血性休克模型，在此基礎(chǔ)上初步探討失血性休克不同液體復(fù)蘇后脾臟中MDSCs數(shù)量變化，篩選出對(duì)失血性休克免疫抑制逆轉(zhuǎn)作用較強(qiáng)的復(fù)蘇液體并指導(dǎo)以后的干預(yù)治療。 對(duì)象及方法 1.72只SD大鼠，隨機(jī)分成4組，，假手術(shù)組（Sham組）、高滲氯化鈉復(fù)蘇組（HRS組）、乳酸林格氏液復(fù)蘇組（LRS組）、晶膠液復(fù)蘇組（LCRS組），Sham組僅接受麻醉，置管及動(dòng)脈監(jiān)測(cè)，不予放血，其余組大鼠建立失血性休克模型。 2.失血性休克模型，參照Chaudry方法建立失血性休克模型，快速放血至已肝素化的儲(chǔ)血瓶?jī)?nèi)，在10分鐘內(nèi)將平均動(dòng)脈壓降至40mmHg為目標(biāo)，期間通過(guò)調(diào)節(jié)放血速度控制血壓。 3.模型制作成功后，分別以不同的液體(高滲氯化鈉液，乳酸林格氏液，晶膠液)進(jìn)行復(fù)蘇，復(fù)蘇后12，24，48小時(shí)分別處死（頸椎脫位法處死大鼠）各組大鼠6只（包括假手術(shù)組），取脾臟組織。 4.將脾臟組織制作成細(xì)胞懸液，His48、CD11bc試劑雙標(biāo)后用流式細(xì)胞儀檢測(cè)各組脾臟中MDSCs，計(jì)算MDSCs的比例變化。 結(jié)果 1.參照Chaudry法成功建立失血性休克模型，達(dá)到平均動(dòng)脈壓40mmHg的要求。 2.假手術(shù)組在三個(gè)不同時(shí)間點(diǎn)脾臟中MDSCs數(shù)量變化在統(tǒng)計(jì)學(xué)上無(wú)明顯差異。 3.相比于假手術(shù)組，失血性休克復(fù)蘇后MDSCs數(shù)量均有不同程度的升高，差異明顯，有統(tǒng)計(jì)學(xué)意義，其數(shù)量變化有先升后降的趨勢(shì)。 4.三種不同的復(fù)蘇液體，在三個(gè)不同時(shí)間點(diǎn)以高滲鹽組復(fù)蘇后MDSCs數(shù)量升高程度相對(duì)較小。12h時(shí)，與晶膠液相比，差異不明顯，與乳酸林格氏液比，差異顯著。但在24h,48h同另兩組相比，具有統(tǒng)計(jì)學(xué)意義。 結(jié)論 1.用Chaudry法建立的失血性休克模型，操作簡(jiǎn)便，易于復(fù)制，能反映失血性休克的發(fā)病過(guò)程和臨床特征。 2.假手術(shù)組機(jī)體損傷較小，MDSCs變化無(wú)統(tǒng)計(jì)學(xué)差異，說(shuō)明實(shí)驗(yàn)動(dòng)物通過(guò)自我保護(hù)反應(yīng)，維持機(jī)體免疫平衡。 3.失血性休克復(fù)蘇后，機(jī)體的免疫功能早期受到不同程度的抑制，隨時(shí)間變化逐漸恢復(fù)。 4.相對(duì)于乳酸林格氏液晶膠液，高滲氯化鈉液復(fù)蘇失血性休克，能有效抑制脾臟中MDSCs數(shù)量，減輕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.
【學(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R459.7

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