七氟烷对失血性休克猪肠屏障损伤的保护作用及对肠粘膜AQP8的影响

发布时间：2018-04-13 00:35

  本文选题：肠 + 失血性休克　； 参考：《大连医科大学》2014年硕士论文

【摘要】：目的： 通过观察七氟烷对巴马小型猪失血性休克肠屏障功能及肠黏膜水通道蛋白8（Aquaporin8，AQP8）表达的影响，从而探讨七氟烷对其保护作用及可能的机制。 方法： 巴马小型猪32头，随机分为4组，每组8头，分别为对照组（S组）、失血性休克组（HS组）、七氟烷预处理组（Pre/Sev组）及七氟烷后处理组（Post/Sev组）。实验动物术前禁食禁饮8h，给予丙泊酚3mg/kg实施麻醉。S组麻醉后经股动脉和颈内静脉置管；HS组麻醉置管后建立失血性休克模型；Pre/Sev组麻醉置管后给予2%七氟烷吸入30min后建立失血性休克模型；Post/Sev组于麻醉置管建立失血性休克模型成功后给予2%七氟烷吸入30min。各实验组均于麻醉前（T0）、失血性休克后30min（T1）、1h（T2）、1.5h（T3）、2h（T4）、3h（T5）、4h（T6）时间点于颈内静脉采取外周血、记录血流动力学指标并且测定心输出量和温度；ELISA法检测血清肿瘤坏死因子（TNF-a）、白介素6(IL-6)、D-乳酸、肠脂肪酸结合蛋白(I-FABP)的含量；失血性休克4h后放血处死实验动物取肠组织制作病理切片，HE染色观察各组病理组织学变化；用干湿比法计算肠组织的含水量；并且取下腔静脉血、肠系膜淋巴结(MLN)、肝、肺、肾、脾组织进行细菌培养，计算远隔器官移位频率；ELISA法检测AQP8表达的变化。采用SPSS19.0软件进行统计学分析，实验数据以均数±标准差(X±s)表示，组间比较采用单因素方差分析，组内比较采用重复测量设计的方差分析，两变量间相关性采用Pearson相关分析，以P0.05为差异有统计学意义。 结果： 1、病理组织学观察光镜显示，S组肠道黏膜未见明显变化；HS组肠道黏膜损伤严重，可见肠粘膜出血，肠粘膜层炎细胞浸润，肠粘膜上皮细胞坏死； Pre/Sev组和Post/Sev组损伤明显减轻，仅见轻度粘膜层腺体扩张，中度的上皮层和固有层分离，上皮下间隙轻度水肿，少许的炎细胞浸润。 2、血清IL-6和TNF-α含量的变化：与S组相比，HS组、Pre/Sev组和Post/Sev组IL-6和TNF-α含量均显著升高，差异有统计学意义（P0.05）。与HS组相比较，Pre/Sev组和Post/Sev组IL-6和TNF-α含量均显著降低，差异有统计学意义（P0.05）。Pre/Sev组和Post/Sev组比较，IL-6和TNF-α含量差异无统计学意义（P＞0.05）。 3、血清D-乳酸和I-FABP含量的变化：与S组相比，HS组、Pre/Sev组和Post/Sev组D-乳酸和I-FABP含量均显著升高，差异有统计学意义（P0.05）；与HS组相比较，Pre/Sev组和Post/Sev组D-乳酸和I-FABP含量明显降低，差异有统计学意义（P0.05）；Pre/Sev组与Post/Sev组相比较，D-乳酸和I-FABP含量差异无统计学意义（P＞0.05）。 4、远隔脏器细菌移位率：与S组相比，HS组、Pre/Sev组和Post/Sev组远隔脏器细菌移位率显著增高，差异有统计学意义（P0.05）；与HS组相比， Pre/Sev组和Post/Sev组远隔脏器细菌移位频率明显降低，差异有统计学意义（P0.05）；Pre/Sev组和Post/Sev组间差异无统计学意义（P＞0.05）。 5、肠黏膜AQP8表达的变化：与S组相比，HS组、Pre/Sev组和Post/Sev组肠黏膜AQP8表达显著增高，差异有统计学意义（P0.05）；与HS组相比， Pre/Sev组和Post/Sev组肠黏膜AQP8含量明显降低，差异有统计学意义（P0.05）；Pre/Sev组和Post/Sev组间差异无统计学意义（P＞0.05）。 6、肠组织含水量的变化：与S组相比，HS组、Pre/Sev组和Post/Sev组肠组织含水量显著增高，差异有统计学意义（P0.05）；与HS组相比， Pre/Sev组和Post/Sev组肠组织含水量明显降低，差异有统计学意义（P0.05）；Pre/Sev组和Post/Sev组间差异无统计学意义（P＞0.05）。 7、失血性休克组空肠、回肠、结肠肠黏膜AQP8表达变化与肠组织含水量进行相关性分析，结果显示，二者之间呈正相关（R2=0.841，0.822，，0.890， P0.05）。 8、失血性休克组血清D-乳酸、I-FABP浓度的变化，与血清炎性因子进行相关分析，结果显示血清TNF-α浓度与D-乳酸、I-FABP浓度的变化呈正相关（R2=0.642，0.697， P0.05），血清IL-6浓度与D-乳酸、I-FABP浓度的变化呈正相关（R2=0.179，0.856， P0.05）。 结论： 1、AQP8在失血性休克肠黏膜损伤中参与肠道组织水肿的形成； 2、七氟烷可以减轻失血性休克引起的肠黏膜屏障损伤； 3、七氟烷对失血性休克肠屏障损伤的保护机制可能与下调AQP8表达，减轻 肠黏膜水肿及抑制炎性反应有关。
[Abstract]:Purpose :

Objective To investigate the effect of heptafluoroalkane on intestinal barrier function and expression of Aquaporin8 ( AQP8 ) in the intestinal mucosal water channel protein 8 ( AQP8 ) of the small porcine hemorrhagic shock .

Method :

Thirty - two groups were randomly divided into four groups : control group ( group S ) , hemorrhagic shock group ( HS group ) , pre / Sev group ( Pre / Sev group ) and group ( Post / Sev group ) .
The hemorrhagic shock model was established after anesthesia with HS group .
The hemorrhagic shock model was established after the Pre / Sev group was intuitized with 2 % of 7 - fluoroalkane after inhalation for 30 min .
The hemodynamic parameters were recorded and the cardiac output and temperature were measured before anesthesia ( T0 ) , 30min after hemorrhagic shock ( T1 ) , 1h ( T2 ) , 1.5h ( T3 ) , 2h ( T4 ) , 3h ( T5 ) and 4h ( T6 ) , respectively .
Serum tumor necrosis factor ( TNF - a ) , interleukin - 6 ( IL - 6 ) , D - lactic acid and intestinal fatty acid binding protein ( I - FABP ) were detected by ELISA .
After 4 hours of hemorrhagic shock , the rats were sacrificed after hemorrhagic shock , pathological sections were prepared from intestinal tissue , and the pathological changes of each group were observed by HE staining ;
calculating the water content of the intestinal tissue by a dry - wet - ratio method ;
and taking inferior vena cava blood , mesenteric lymph node ( MLN ) , liver , lung , kidney and spleen tissue to carry out bacterial culture to calculate distant organ shift frequency ;
The changes of AQP8 expression were detected by ELISA . SPSS 10.0 software was used for statistical analysis . The experimental data was expressed by mean 卤 standard deviation ( X 卤 s ) . A single - factor analysis was used to compare the differences between the groups . Pearson correlation was used to analyze the correlation between the two variables .

Results :

1 . Pathohistological observation showed that there was no obvious change in intestinal mucosa of S group .
The intestinal mucosal lesion was severe in HS group , and intestinal mucosal bleeding , infiltration of intestinal mucosal layer inflammatory cells and necrosis of intestinal epithelial cells were observed .
The pre / Sev group and Post / Sev group were significantly reduced , only mild mucosal layer gland dilatation , moderate upper cortical layer and intrinsic layer separation , mild edema of the upper subcutaneous gap , and little inflammatory cell infiltration .

Compared with group S , the levels of IL - 6 and TNF - 伪 in Pre / Sev group and Post / Sev group were significantly lower than those in HS group ( P0.05 ) . There was no significant difference between Pre / Sev group and Post / Sev group ( P > 0.05 ) .

3 . Changes of serum D - lactic acid and I - FABP content : Compared with group S , the contents of D - lactic acid and I - FABP in HS group , Pre / Sev group and Post / Sev group increased significantly ( P0.05 ) .
Compared with HS group , the contents of D - lactic acid and I - FABP decreased significantly in Pre / Sev group and Post / Sev group ( P0.05 ) .
Compared with the Post / Sev group , the contents of D - lactic acid and I - FABP were not statistically significant ( P > 0.05 ) .

4 . Bacterial translocation rate of the distant organs : Compared with the S group , the rate of bacterial translocation was significantly higher in HS group , Pre / Sev group and Post / Sev group than in S group ( P0.05 ) .
Compared with HS group , the frequency of bacterial translocation between Pre / Sev group and Post / Sev group was significantly lower than that of HS group ( P0.05 ) .
There was no significant difference between Pre / Sev group and Post / Sev group ( P > 0.05 ) .

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