内源性一氧化碳对感染性休克时肺脏和肾脏损伤的保护作用及其机制初探

发布时间：2018-02-01 09:12

本文关键词： 一氧化碳血红素氧合酶内毒素肺肾炎症　出处：《河北医科大学》2005年硕士论文　论文类型：学位论文

【摘要】：目的:腹腔感染所致的感染性休克是一种临床常见的病理过程,发病急骤,并发症多,可累及机体多个器官,甚至导致多器官功能障碍综合征(multiple organ dysfunctionsyndrome,MODS)。而肺脏和肾脏是经常受累的器官,主要为内素素诱导诱导炎症介质失控性释放,从而引起机体器官的损伤,但其具体的作用机制尚未完全明了,氧化应激可能是肺和肾损伤的主要机制之一。已有研究表明,一氧化碳(carbon monoxide,CO)可抑制机体炎症因子的产生,对抗氧自由基损伤,但对盲肠结扎穿孔(cecal ligation andpuncture,CLP)导致感染性休克时肺和肾损伤是否也具有保护作用尚有待探讨。本研究使用CLP 的方法建立感染性休克模型,通过应用血红素氧合酶(heme oxygenase,HO)诱导剂氯血红素(Hemin)和抑制剂锌原卟啉(Zinc protoporphyrin,ZnPP)调节HO-1 活性来控制内源性CO 的生成,探讨内源性CO 对肺和肾损伤的保护作用及其可能机制。方法:实验用雄性Sprague Dawley(SD)大鼠,体重200～250 克,随机分为四组,每组24 只。(1)假手术组:腹腔注射25%乌拉坦4mg/kg 麻醉,行腹部正中切口,于盲肠根部穿线而不结扎,逐层缝合;(2)CLP 组:开腹后结扎盲肠根部,在肠系膜相对的盲肠浆膜面用18 号针头穿刺两个孔(相距1 厘米),逐层缝合;(3)CLP+Hemin 组:手术前10min 经舌下静脉注射Hemin(10mg/kg),其余操作同CLP 组;(4)CLP+ZnPP 组:手术前10min 舌下静脉注射ZnPP(10mg/kg),其余操作同CLP 组。各组分别于模型成立后2h、4h、6h 从右颈静脉插管至右心房,分离左颈静脉插管至主动脉根部,采集血液分别代表出肺血(Out-going pulmonaryblood,OPB)和入肺血(In-flowing pulmonary blood,IPB),以检测其碳氧血红蛋白(carboxyhemoglobin,COHb)水平,以此代表CO 含量,经颈静脉取血液标本后继续放血处死动物,取肺、肾组织标本,将血液标本离心后取上清液,将肺、肾组织标本制备匀浆,以检测血液及肺、肾组织中的超氧化物歧化酶(superoxidedismutase,SOD)活性、丙二醛(malondialdehyde,MDA)含量。另外留取肺、肾组织标本,光镜观察其病理学改变,免疫组织化学染色观察其HO-1 的表达及分布。数据用均数±标准差(x±s)表示。单因素方差分析进行组间比较,有显著差异者用最小显著差法进行两两比较,以p0.05 为有显著差异。结果:各组不同时间点OPB 和IPB 的COHb 含量和血液及肺、肾组织匀浆的SOD 活性、MDA 含量。①与同时间点Sham 组相比,CLP 组2h、4h、6hOPB 和IPB 的COHb含量及其差值(OPB-IPB)均显著升高(P 均0.05),且相邻时间点间也有显著差异(P 均0.05),并呈递增趋势,血液及肺、肾组织匀浆的SOD 活性显著降低(P 均0.05 或0.01),相邻时间点间也有显著差异(P 均0.05 或0.01),呈递减趋势,而MDA 含量则显著升高(P 均0.05 或0.01),呈递增趋势;②与同时间点CLP 组相比,CLP+Hemin 组2h、
[Abstract]:Objective: septic shock caused by abdominal infection is a common clinical pathological process. It even leads to multiple organ dysfunctionsyndrome with multiple organ dysfunction syndrome. MODS. Lung and kidney are often involved organs, mainly for endotoxin induced inflammatory mediators release out of control, thus causing organ damage, but its specific mechanism has not been fully understood. Oxidative stress may be one of the main mechanisms of lung and kidney injury. It has been shown that carbon monoxide can inhibit the production of inflammatory factors. Oxygen free radical injury was inhibited, but cecal ligation andpuncture was ligated and perforated by cecum ligation. Whether lung and kidney injury also have protective effect on septic shock. In this study, CLP was used to establish septic shock model. Heme oxygenase of heme oxygenase was used. ) and zinc protoporphyrin zinc protoporphyrin. To investigate the protective effect of endogenous CO on lung and kidney injury and its possible mechanism. Methods: male Sprague Dawley SD rats, weighing 200,250g, were randomly divided into four groups. Sham-operation group (n = 24): intraperitoneal injection of 25% uratan (4 mg / kg) was performed through the median abdominal incision, without ligation at the root of the cecum, and then sutured layer by layer. In the CLP group, the caecum root was ligated after laparotomy, and two holes (1 cm apart, sutured) were punctured on the cecum serosa surface opposite to the mesentery with a needle of 18 (1 cm apart). Hemin group: operation In the first 10 minutes, 10 mg 路kg ~ (-1) 路kg ~ (-1) of Heminine was injected into the sublingual vein. The rest of the patients were treated with CLP. ZnPP group: 10 minutes before operation, 10 mg / kg of ZnPP was injected into the sublingual vein, and the other operation was the same as that in the CLP group. Each group was treated with CLP for 4 hours after the establishment of the model. The left jugular vein was intubated to the root of the aorta at 6 h from the right jugular vein to the right atrium. The blood collected from the left jugular vein represented the lung blood Out-going pulmonaryblood. In order to detect carboxyhemoglobin (carboxyhemoglobin), OPBand pulmonary blood cells were used to detect carboxyhemoglobin. The level of COHb, which represents the content of CO, continues to be exorcised from the jugular vein to kill the animals, to collect the lung and kidney tissue, to centrifuge the blood sample and to take the supernatant and the lung. The homogenate of kidney tissue was prepared to detect the activity of superoxide dismutase (SOD) in blood, lung and kidney. The content of malondialdehyde (MDA) and the pathological changes of lung and kidney were observed under light microscope. The expression and distribution of HO-1 were observed by immunohistochemical staining. The data were expressed as mean 卤standard deviation (x 卤s). Single factor ANOVA was used to compare the two groups, and the least significant difference was used to compare the two groups, with p0.05 as the significant difference. Results: the COHb content of OPB and IPB and the activity of SOD in kidney homogenate were compared with those in Sham group at different time points. The COHb content and the difference of OPB and IPB in CLP group were significantly higher than those in control group (P < 0.05). There were also significant differences between adjacent time points (P 0.05 or 0.01). The activity of SOD in blood, lung and kidney homogenate decreased significantly (P 0.05 or 0.01). There were also significant differences between adjacent time points (P 0.05 or 0.01), but the content of MDA increased significantly (P 0.05 or 0.01). 2Compared with the CLP group at the same time point for 2 h,
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R363

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