本文選題：急性肺損傷 + 內毒素��； 參考：《中國人民解放軍醫(yī)學院》2013年博士論文

【摘要】：目的：觀察角質化細胞生長因子(KGF)對大鼠急性肺損傷(ALI)模型的防護作用,觀察動脈血氣、肺系數(shù)、肺組織細胞標志物的變化及MicroRNA的差異表達,為KGF對ARDS/ALI防護作用的機制研究提供新的思路。 方法：本實驗分為三個部分： 實驗一：KGF對LPS致大鼠ALI/ARDS的干預作用。18只SD大鼠按體重隨機分為A組：為正常對照組：B組：為ALI/ARDS模型組,經尾靜脈按6mg/kg注射脂多糖(LPS); C組：為KGF干預組,尾靜脈注射LPS前72h,經氣道按5mg/kg滴注KGF。尾靜脈注射LPS8h后觀察各組大鼠的血氣分析、肺系數(shù)和肺組織病理學變化。 實驗二：KGF對LPS所致大鼠ALI/ARDS肺組織勻漿中KL-6、AQP5及ACE的影響。實驗分組同實驗一。通過Western Blot觀察不同實驗組大鼠肺組織勻漿中KL-6、水通道蛋白5(AQP5)和血管緊張素轉化酶(ACE)含量的變化。 實驗三：KGF對LPS所致肺損傷microRNA表達的影響。6只SD大鼠隨機分為2組,每組3只。LPS組：為ALI/ARDS模型組,經尾靜脈按6mg/kg注射脂多糖(LPS); KGF組：為KGF干預組,尾靜脈注射LPS前72h,經氣道按5mg/kg滴注KGF。尾靜脈注射LPS8h后利用高通量芯片對比LPS組和KGF組大鼠肺組織中microRNA的差異表達,并通過real-time-PCR進行驗證。 結果： 實驗一：與A組比較,其他各組大鼠PaO2、PaO2/FiO2均有下降(P0.01),各染毒組大鼠PaCO2較A組均有升高,有極顯著差異(P0.01),與B組相比,C組大鼠PaO2、PaCO2及PaO2/FiO2的改善顯著(P0.01或P0.05)；B組和C組大鼠肺濕干比、肺含水量較A組顯著增高(P0.01或P0.05)；光鏡下觀察可見B組大鼠肺組織肺泡壁高度增生肥厚,可見纖維化區(qū)域,同時又大量淋巴細胞浸潤和少量紅細胞浸潤,C組較B組明顯改善。 實驗二：與A組相比,B組和C組大鼠肺組織勻漿中KL-6含量均有顯著增高(P0.05),C組較之B組大鼠肺組織勻漿中KL-6含量也有明顯升高,差別有統(tǒng)計學意義(P0.05)；與A組相比,其他各組大鼠肺組織勻漿中AQP5的含量明顯下降(P0.05),與B組相比,C組大鼠肺組織勻漿中AQP5的含量有升高,但差別無統(tǒng)計學意義(p0.05)；與A組相比,其他各組大鼠肺組織勻漿中ACE含量均有升高,差別有統(tǒng)計學意義(P0.05),C組較B組大鼠肺組織勻漿ACE含量有下降,但兩者之間的差異不顯著(P0.05)。 實驗三：芯片結果顯示兩組間無差異表達的miRNA,通過生物信息學預測和real-time-PCR驗證發(fā)現(xiàn),與LPS組相比,miR-494顯著上調,miR-145和miR-146b顯著下調,差別具有統(tǒng)計學意義(p0.01)。 結論： 1.KGF對LPS致大鼠急性肺損傷具有預防作用,可以顯著增加PaO2、 PaO2/FiO2,降低PaCO2、肺濕干比和含水量,刺激AT-Ⅱ細胞增殖是KGF發(fā)揮作用的可能機制之一。 2.KGF可通過促進AT-Ⅱ細胞增殖,增加AT-Ⅰ細胞,維持肺血管內皮細胞完整性來發(fā)揮其肺損傷防護作用。 3. miR-494、miR-145和miR-146b在KGF組大鼠肺組織的表達有明顯變化,這些miRNA與促進細胞增殖和抗凋亡密切相關,可能是KGF是預防和治療LPS致肺損傷的重要靶點。
[Abstract]:Objective: To observe the protective effect of keratinocyte growth factor (KGF) on acute lung injury (ALI) model in rats, observe the changes of arterial blood gas, lung coefficient, change of lung tissue cell markers and the differential expression of MicroRNA, and provide a new idea for the mechanism of KGF for the protection of ARDS/ALI.
Methods: the experiment was divided into three parts:
Experiment 1: the intervention of KGF on LPS induced ALI/ARDS in rats.18 rats were divided into A group according to weight randomly: the normal control group: the B group: the ALI/ARDS model group, the tail vein in 6mg/kg injection of lipopolysaccharide (LPS), the C group: the tail vein was injected before the tail vein injection, and the meridian was injected into the tail vein to observe each group. Blood gas analysis, lung coefficient and pathological changes of lung tissue in rats.
Experiment two: the effect of KGF on KL-6, AQP5 and ACE in ALI/ARDS lung homogenate of rats induced by LPS. Experimental group and Experiment 1. Observe the changes of KL-6, aquaporin 5 (AQP5) and angiotensin converting enzyme (ACE) in the lung homogenate of different experimental groups by Western Blot.
Experiment three: the effect of KGF on the expression of microRNA in lung injury induced by LPS,.6 rats were randomly divided into 2 groups, each group of 3.LPS groups, ALI/ARDS model group and 6mg/kg injection of lipopolysaccharide (LPS) through 6mg/kg, KGF group: KGF intervention group, caudal vein before LPS. The differential expression of microRNA in lung tissue of rats in group LPS and KGF was verified by real-time-PCR.
Result錛，

本文編號：1936921