【摘要】：目的：探討組氨酸脫羧酶及D-乳酸在絞窄性腸梗阻早期診斷中的應用價值。 方法：將雄性Wistar大鼠分為對照組10只，實驗組20只。實驗組通過結扎腸管制備絞窄性腸梗阻模型，根據(jù)腸梗阻發(fā)生時間分為A、B兩個亞組，A組為絞窄性腸梗阻1h，B組為絞窄性腸梗阻3h。光鏡下觀察各組大鼠肝、小腸組織病理學變化，依據(jù)Park/Chiu腸組織損傷評級標準判定腸壁損傷程度。ELISA法檢查各組大鼠血清HDC和D-乳酸濃度。利用RT-PCR和實時熒光定量PCR檢測肝、小腸組織中HDC mRNA表達水平。 結果：光鏡下小腸組織損傷評分實驗組高于對照組。對照組血清HDC濃度為9.973.64pg/ml，發(fā)生絞窄性腸梗阻1h后血清HDC濃度為83.9727.69pg/ml，，明顯高于對照組（P0.01），而3h后血清HDC濃度較之1h水平進一步增高，達到143.2147.15pg/ml（P0.05）。對照組血清D-乳酸濃度為1.111.79ng/ml，發(fā)生絞窄性腸梗阻3h后血清D-乳酸濃度為10.3614.91ng/ml，和對照組相比雖有增高的趨勢，但差異無統(tǒng)計學意義（P0.05）。RT-PCR電泳顯示絞窄性腸梗阻3h時，小腸組織中HDC條帶亮度明顯強于對照組。實時熒光定量PCR結果顯示，絞窄性腸梗阻后肝組織HDC mRNA表達水平A組是對照組的1.34倍，B組是對照組的2.21倍， A組和對照組無差異（P0.05）， B組和對照組比較明顯增高（P0.05）。絞窄性腸梗阻后小腸組織HDC mRNA表達水平A組是對照組的1.81倍，B組是對照組的8.02倍，A組和對照組無差異（P0.05），B組和A組及對照組比較均顯著增高（P0.01）。 結論：1、絞窄性腸梗阻早期肝和小腸組織HDC mRNA表達水平明顯增高。2、絞窄性腸梗阻早期血清HDC濃度顯著增高。3、小面積腸缺血早期血清D-乳酸濃度無明顯改變。4、HDC基因的表達水平可作為絞窄性腸梗阻早期診斷指標之一。5、HDC基因的高表達預示腸梗阻的嚴重程度。
[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.
【學位授予單位】：吉林大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R656

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