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Received:February 22, 2022 Published Online:September 20, 2022
Received:February 22, 2022 Published Online:September 20, 2022
中文摘要: 目的 通过中药方剂灌胃建立功能性消化不良 (FD) 大鼠模型,研究其小肠Cajal间质细胞(ICC)中凋亡相关蛋白Caspase-3蛋白及mRNA的表达,探讨Caspase-3在FD大鼠小肠ICC中的表达及意义。方法 选用80只SPF级的Wistar成年大鼠,雌雄各半,随机分为对照组(n=30)和FD模型组(n=50)。对照组大鼠生理盐水灌胃,FD模型组用中药方剂灌胃建立FD大鼠动物模型。喂养和造模过程中观察并测量两组大鼠的一般情况和体重变化,造模后测定尿D-木糖排泄率和小肠推进率,然后颈椎脱臼法处死大鼠,取距幽门远端8 cm处小肠并通过免疫荧光法测定ICC的数量及分布,透射电镜观察ICC形态学变化,Western blot及逆转录聚合酶链反应(RT-PCR)法检测ICC中Caspase-3蛋白及mRNA的表达。结果 与对照组相比,FD模型组大鼠的平均体重明显下降,体重增长减缓,尿D-木糖排泄率明显降低,差异均有统计学意义(P<0.05),说明模型组成功建立了FD模型。与对照组相比,FD 模型组的小肠 ICC体积变小,细胞器减少,细胞质肿胀甚至空泡化,与周围平滑肌细胞和神经纤维末梢之间的连接结构存在不同程度的紊乱及破坏。激光共聚焦显微镜荧光定量分析显示,FD模型组大鼠小肠组织内 ICC 的分布较对照组明显减少,差异有统计学意义(P<0.05)。与对照组相比,FD 模型组的大鼠小肠ICC中 Caspase-3的蛋白和mRNA相对表达量明显增加,差异有统计学意义(P<0.05)。结论 大鼠 ICC 的形态结构变化在FD发病中发挥作用,Caspase-3高表达导致的小肠ICC凋亡是FD症状的可能原因。
Abstract:Objective To explore the expression and significance of apoptosis related protein caspase-3 in interstitial cell of Cajal (ICC) in small intestine of rats with functional dyspepsia (FD). Methods Eighty adult Wistar rats of specific pathogen free(SPF)grade (half male and half female) were randomly divided into control group (n=30) and FD model group (n=50). Rats in control group were perfused with normal saline, and rats in FD model group were perfused with Chinese medicine to establish FD rat model. The general state and weight changes of rats were observed during feeding and modeling. After modeling, urinary D-xylose excretion rate and intestinal propulsive rate were determined in each group, and the rats were killed by cervical dislocation. The small intestine was taken from 8 cm distal to the pylorus to measure the number and distribution of ICC by immunofluorescence and to observe the morphological changes of ICC by transmission electron microscope (TEM). Western blot and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expressions of Caspase-3 protein and mRNA in ICC. Results Compared with those in control group, the average body weight and the urinary D-xylose excretion rate significantly reduced, and ICC presented with smaller volume, fewer organelles, swelling and even vacuolization of cytoplasm, as well as varying degrees of disorder and destruction of the connecting structure with surrounding smooth muscle cells and nerve fiber terminals in FD model group(P<0.05). Confocal fluorescence microscopy showed that the distribution of ICC in the small intestine of FD rats was significantly lower than that in control group (P<0.05). Caspase-3 protein and mRNA expression levels in ICC significantly increased in group B (P<0.05). Conclusion Morphological and structural changes of ICC in rats play a role in the pathogenesis of FD. Apoptosis of ICC in small intestine induced by high expression of Caspase-3 is a possible cause for functional dyspepsia.
keywords: Functional dyspepsia Apoptosis related Caspase-3 Interstitial cell of Cajal Urinary D-xylose
文章编号: 中图分类号:R57 文献标志码:B
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