大鼠绞窄性肠梗阻早期组氨酸脱羧酶和D-乳酸的变化

发布时间：2019-02-11 07:45

【摘要】：目的：探讨组氨酸脱羧酶及D-乳酸在绞窄性肠梗阻早期诊断中的应用价值。方法：将雄性Wistar大鼠分为对照组10只，实验组20只。实验组通过结扎肠管制备绞窄性肠梗阻模型，根据肠梗阻发生时间分为A、B两个亚组，A组为绞窄性肠梗阻1h，B组为绞窄性肠梗阻3h。光镜下观察各组大鼠肝、小肠组织病理学变化，依据Park/Chiu肠组织损伤评级标准判定肠壁损伤程度。ELISA法检查各组大鼠血清HDC和D-乳酸浓度。利用RT-PCR和实时荧光定量PCR检测肝、小肠组织中HDC mRNA表达水平。结果：光镜下小肠组织损伤评分实验组高于对照组。对照组血清HDC浓度为9.973.64pg/ml，发生绞窄性肠梗阻1h后血清HDC浓度为83.9727.69pg/ml，，明显高于对照组（P0.01），而3h后血清HDC浓度较之1h水平进一步增高，达到143.2147.15pg/ml（P0.05）。对照组血清D-乳酸浓度为1.111.79ng/ml，发生绞窄性肠梗阻3h后血清D-乳酸浓度为10.3614.91ng/ml，和对照组相比虽有增高的趋势，但差异无统计学意义（P0.05）。RT-PCR电泳显示绞窄性肠梗阻3h时，小肠组织中HDC条带亮度明显强于对照组。实时荧光定量PCR结果显示，绞窄性肠梗阻后肝组织HDC mRNA表达水平A组是对照组的1.34倍，B组是对照组的2.21倍， A组和对照组无差异（P0.05）， B组和对照组比较明显增高（P0.05）。绞窄性肠梗阻后小肠组织HDC mRNA表达水平A组是对照组的1.81倍，B组是对照组的8.02倍，A组和对照组无差异（P0.05），B组和A组及对照组比较均显著增高（P0.01）。结论：1、绞窄性肠梗阻早期肝和小肠组织HDC mRNA表达水平明显增高。2、绞窄性肠梗阻早期血清HDC浓度显著增高。3、小面积肠缺血早期血清D-乳酸浓度无明显改变。4、HDC基因的表达水平可作为绞窄性肠梗阻早期诊断指标之一。5、HDC基因的高表达预示肠梗阻的严重程度。
[Abstract]:Objective: to evaluate the value of histidine decarboxylase and D-lactic acid in the early diagnosis of strangulated intestinal obstruction. Methods: male Wistar rats were divided into control group (n = 10) and experimental group (n = 20). The model of strangulated intestinal obstruction was established by ligation of intestinal tube in the experimental group. According to the time of occurrence of intestinal obstruction, the experimental group was divided into two subgroups: group A was strangulated intestinal obstruction for 1 h and group B was strangulated intestinal obstruction for 3 h. The histopathological changes of liver and small intestine were observed under light microscope, and the degree of intestinal wall injury was determined according to the criteria of Park/Chiu intestinal tissue injury. The serum HDC and D-lactic acid concentrations in each group were detected by ELISA method. RT-PCR and real-time quantitative PCR were used to detect the expression of HDC mRNA in liver and small intestine. Results: the injury score of small intestine in the experimental group was higher than that in the control group under light microscope. The serum HDC concentration in the control group was 9.973.64 PG / ml, and the serum HDC level was 83.9727.69 PG / ml after 1 hour of strangulated intestinal obstruction, which was significantly higher than that in the control group (P0.01). 143.2147.15pg/ml was achieved (P0.05). The serum D- lactate concentration in the control group was 1.111.79 ng / ml, and that in the control group was 10.3614.91 ng / ml after 3 hours of strangulated intestinal obstruction. But there was no significant difference (P0.05). RT-PCR electrophoresis showed that the brightness of HDC bands in small intestine tissue was significantly stronger than that in control group at 3 h after strangulation. The results of real-time fluorescence quantitative PCR showed that the expression level of HDC mRNA in group A was 1.34 times higher than that in control group and 2.21 times in group B after strangulated intestinal obstruction. There was no difference between group A and control group (P0.05). Group B and control group were significantly higher (P0.05). The expression level of HDC mRNA in small intestine after strangulated intestinal obstruction was 1.81 times higher in group A than that in control group and 8.02 times in group B, but there was no difference between group A and control group (P0.05). Group B was significantly higher than group A and control group (P0.01). Conclusion: 1. The expression of HDC mRNA in liver and small intestine was significantly increased in the early stage of strangulated intestinal obstruction, the serum HDC level was significantly increased in the early stage of strangulated intestinal obstruction, and the level of serum D-lactic acid was not changed in the early stage of small area intestinal ischemia. The expression level of HDC gene can be used as one of the early diagnostic markers of strangulated intestinal obstruction. The high expression of HDC gene can predict the severity of intestinal obstruction.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R656

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