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目的探讨血清组氨酸脱羧酶(HDC)及D-乳酸浓度在大鼠绞窄性肠梗阻早期诊断中的应用价值。方法将30只雄性Wistar大鼠随机分为空白对照组10只和实验组20只。实验组大鼠通过结扎肠管以制备绞窄性肠梗阻模型,并随机分为肠梗阻1 h组和肠梗阻3 h组,每组10只。空白对照组大鼠仅行开关腹手术。取回肠组织,光镜下观察大鼠回肠组织的病理学变化,并判定肠壁损伤程度;同时行心脏采血,以酶联免疫吸附分析法(ELISA法)检测血清HDC和D-乳酸浓度;再利用实时荧光定量PCR法(RT-PCR法)检测回肠组织中HDC m RNA的表达水平。比较3组大鼠的上述指标。结果光镜下,空白对照组大鼠回肠组织损伤程度评分的中位数为0分(0~1分),肠梗阻1 h组为2分(2~3分),肠梗阻3 h组为5分(4~5分),空白对照组、肠梗阻1 h组和肠梗阻3 h组大鼠回肠组织的损伤程度评分依次增高(P<0.01)。空白对照组大鼠血清HDC浓度的中位数为10.5 pg/m L(4.60~17.18 pg/m L),肠梗阻1 h组为87.93 pg/m L(41.33~119.03 pg/m L),肠梗阻3 h组为150.67 pg/m L(67.33~198.14 pg/m L),空白对照组、肠梗阻1 h组和肠梗阻3 h组大鼠的血清HDC浓度依次增高(P<0.05)。空白对照组大鼠血清D-乳酸浓度的中位数为0 ng/m L(0~3.90 ng/m L),肠梗阻1 h组为0 ng/m L(0~15.63 ng/m L),肠梗阻3 h组为4.92 ng/m L(0~48.13 ng/m L),3组大鼠的血清D-乳酸浓度比较差异无统计学意义(P>0.05)。RT-PCR结果显示,肠梗阻3 h组大鼠回肠组织中HDC m RNA的表达水平的中位数为7.81(7.05~8.39),高于肠梗阻1 h组的1.77(1.74~1.94)和空白对照组的0.97(0.88~1.15),P<0.01;而肠梗阻1 h组和空白对照组间比较差异则无统计学意义(P>0.05)。结论血清HDC浓度可作为绞窄性肠梗阻的早期诊断指标之一,而血清D-乳酸浓度在肠缺血早期无显著改变。
Objective To investigate the value of serum HDAC and D-lactate in the early diagnosis of strangulated intestinal obstruction in rats. Methods Thirty male Wistar rats were randomly divided into blank control group (n = 10) and experimental group (n = 20). Rats in the experimental group were prepared by straining the intestine to make strangulated intestinal obstruction model and randomly divided into 1 h group of intestinal obstruction and 3 h of intestinal obstruction group, 10 rats in each group. Blank control group rats only open the abdominal surgery. The intestinal tissues were removed and pathological changes of ileum were observed under light microscope, and the degree of damage of the intestinal wall was determined. At the same time, blood samples were taken from the heart and serum HDC and D-lactic acid concentrations were detected by enzyme linked immunosorbent assay (ELISA) Real-time quantitative PCR (RT-PCR) was used to detect the expression of HDC m RNA in ileum. The above indexes of 3 groups of rats were compared. Results Under the light microscope, the median ileum tissue injury score was 0 (0-1) in the blank control group, 2 (2 ~ 3) in the 1 h group and 1 h in the 3 h group The scores of ileal injury in the control group, 1 h group with intestinal obstruction and 3 h with intestinal obstruction increased successively (P <0.01). The median serum HDC concentration of the blank control group was 10.5 pg / m L (4.60-17.18 pg / m L) and the intestinal obstruction was 87.93 pg / m L (41.33-119.03 pg / m L) The levels of serum HDC in the control group, 1-h group with intestinal obstruction and 3-h group with intestinal obstruction increased successively (P <0.05) after obstruction for 3 h group was 150.67 pg / m L (67.33-198.14 pg / m L) The median concentration of D-lactic acid in the blank control group was 0 ng / m L (0-3.93 ng / m L) and 0 ng / m L (0-15.63 ng / m L) , Intestinal obstruction 3 h group was 4.92 ng / m L (0 ~ 48.13 ng / m L), 3 groups of rats serum D-lactic acid concentrations were no significant difference (P> 0.05). RT-PCR results showed that the median of HDC m RNA expression in ileum of rats with intestinal obstruction 3 h was 7.81 (7.05 ~ 8.39), which was higher than that of intestinal ileus 1 h (1.74 ~ 1.94) and blank 0.97 (0.88-1.15) in the control group, P <0.01; but there was no significant difference between the 1-h group and the blank control group (P> 0.05). Conclusion Serum HDC concentration can be used as one of the early diagnostic indicators of strangulated intestinal obstruction, while serum D-lactate concentration has no significant change in the early stage of intestinal ischemia.