Objective To investigate the autophagy of Cajal mesenchymal cells in the colon Cajal in rats with slow transit constipation,and qualitatively analyze the calcium ion level.Methods Forty healthy Wistar male rats were di vided into control group (n=20,normal rats) and model group (n=20,model rats) by random number table.Compound phenylethylpiperidine was used for preparation on STC rat model,grain number,every grain of waste quality,body quality of rats,the first grain of black were recorded to judge the model successfully established or not.Colon ICCs of rats in two groups were detected bydetection of immunofluorescence assay,and the concentration of calcium ions in ICCs was determined by immunofluorescence assay.Results The control group had good growth during the experiment,and one died in the model group.Daily waste grain number,average body quality after 30 days in model group were less than those in the control group (P ＜ 0.01),the average grain of feces quality was higher than that in the control group (P ＜ 0.05),and the first row of black time was obviously prolonged in the control group (P ＜ 0.05).The rats colon tissue of ICCs in control group were intertwined to form the intensive and precision of the mesh structure,they had obvious cell branch.The number of ICCs in the intestinal tract of the model group was significantly reduced,and the ICCs was thin and the cell shape narrowed.Compared with the control group,the autophagy number of ICCs in the model group increased and showed the apparent autophagy death.In the control group,ICCs were smooth and had multiple directional and irregular cell branches,and intercellular interaction was connected to the network structure.The number of fluorescence spots and fluorescence intensity in the ICCs of STC rats were significantly higher than those in the control group.Conclusion The number of ICCs in STC rats is reduced and autophagy is occurred.In the STC rat model,the free Ca2+ ions in the ICCs cells are elevated,and the increasing of Ca2+ ion may be the cause of the ICCs' autophagy death.%目的 探讨慢传输型便秘(STC)大鼠结肠Cajal间质细胞(ICCs)自噬现象的发生情况,并定性分析其钙离子浓度水平的变化.方法 选用SPF级健康Wistar雄性大鼠40只,按照随机数字表法分为对照组(n=20,正常大鼠)和模型组(n=20,造模大鼠).采用复方苯乙哌啶制备STC大鼠模型,记录大鼠粪便粒数、每粒粪便质量、大鼠体重及首粒黑便排出时间的变化,判定模型成功建立与否.免疫荧光检测两组大鼠结肠ICCs形态变化,透射电镜检测结肠ICCs自噬情况,免疫荧光法测定ICCs中的钙离子浓度.结果 对照组大鼠试验期间均生长良好,模型组1例死亡.模型组大鼠日均粪便粒数及造模第30天模型组平均体质量较对照组明显减少(P＜0.05),平均每粒粪便质量较对照组升高(P＜0.05),首次排黑便时间较对照组明显延长(P＜0.05).对照组大鼠结肠组织中的ICCs细胞相互交织相连形成了密集且精密的网状结构,存在条形分支.模型组大鼠肠道内ICCs细胞数量明显减少,ICCs交互网络稀薄,细胞变窄变细.与对照组比较,模型组中ICCs细胞自噬泡数量上升且表现出明显细胞自噬性死亡.对照组大鼠中ICCs细胞胞体平滑且细胞分叉,细胞间连接成网络结构.STC大鼠ICCs中荧光斑点数量及荧光强度与对照组比较,明显升高.结论 STC大鼠肠道内ICCs的数量减少是发生了细胞自噬现象.STC大鼠模型中ICCs细胞内游离Ca2+升高,Ca2+升高可能是导致ICCs自噬性死亡的原因.
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