苦杏仁对老年慢传输型便秘大鼠结肠Cajal间质细胞的影响

Effects of bitter almond on interstitial cells of Cajal in old rats with slow-transit constipation

目的观察苦杏仁对老年慢传输型便秘大鼠结肠Cajal间质细胞影响，探讨苦杏仁的通便机制。方法45只老年SD大鼠随机分为空白对照组、模型对照组、苦杏仁实验组，各15只。应用复方苯乙哌啶（地芬诺酯）饲大鼠建立慢传输型便秘大鼠模型。苦杏仁实验组大鼠经灌胃给予苦杏仁煎剂，空白对照组和模型对照组大鼠经灌胃给予相应量的生理盐水。10d后采用活性炭灌胃法检测大鼠肠道推进率，提取结肠组织，采用免疫组化、蛋白免疫印迹（Westernblot）试验和反转录聚合酶链反应（RT-PCR），检测Cajal间质细胞在结肠组织的分布和数量变化、c—kit蛋白以及c—kit基因的表达。结果模型对照组与空白对照组相比肠道推进率下降（47．7±1．7）％比（68．6±6．0）％（Pd0．01），c—kit表达阳性面积降低[（638．0土23．5）μm^2比（723．7士30．8）μm^2，（P〈0．05）]，c—kit基因的表达量明显降低0．3±0．1比1．0±0．1（P〈0．05），c—kit蛋白的表达明显降低0．2±0．2比0．5±0．2（P〈0．05）；苦杏仁实验组与模型对照组相比肠道推进率上升，（63．4±3．3）％比（47．7±1．7）％（Pd0．01），c—kit基因的表达上调0．9±0．2比0．3±0．1（P〈0．01），c—kit蛋白的表达上调0．5士0．3比0．2±0．2（P〈0.01）。结论苦杏仁能促进老年慢传输型便秘大鼠结肠组织中c—kit的表达，修复部分慢传输型便秘大鼠结肠组织中Cajal间质细胞，从而促进慢传输型便秘大鼠的肠道蠕动。

Objective To study effects of bitter almond on interstitial cells of Cajal （ICC） in old rats with slow-transit constipation （STC） and analyze mechanism of the gut purge of bitter almond. Methods Forty-five old SD rats were randomly divided into blank control group,model control group, bitter almond-treated group randomly （n= 15, each）. Compound phenanthroline was used to build the model of STC model. The bitter almond-treated group was given with bitter almond apozem,the other two groups were given with the same amount of saline. After 10 days, the carbon powder propulsion rates in the intestine were measured and specimens were taken. The ICC changes in terms of the number and distribution were observed by immunohistochemical method. The protein and mRNA expressions of c-kit in ICC were measured by Western blot and reverse transcription-polymerase chain reaction（RT-PCR）. Results Compared with blank control group [（ 68.6 ± 6.0）%], the carbon powder propulsion rates in the intestine in model control group [（47.7 ± 1. 7）%] were declined （P〈 0.01）. The areas of c-kit positive cell in colon were declined [ （ 638.0 ± 23.5 ） μm^2 vs. （ 723.7 ±30.8 ） μm^2]in model vs. blank control （P〈0.05）. The mRNA expression of c-kit gene in ICC was declined [（0.3±0.1） vs. （1.0±0.1）]（P〈0.05） ,the expression of c-kit protein in ICC were declined[（0.2± 0. 2） vs. （0.5 ±0.2） ] （P〈 0.05 ）. Compared with model control group [ （ 47.7±1.7 ） % ], the carbon powder propulsion rates in the intestine in bitter almond group were increased[（63.4±3.3）% vs. ] （P 〈0.01）. The mRNA expression of c-kit gene in ICC were increased in bitter almond group （0.9±0. 2） vs model control group [（0.3 ± 0.1 ） ] （P 〈 0.01 ）. The expression of c-kit protein in ICC were increased in bitter almond group[（0.5±0.3） vs. in model control （0.2 ±0.2）] （P〈0.01）.Conclusions The bitter almond can promote the expression of c kit mRNA and protein in colonic tissue of old STC rat interstitial and repair a fraction of ICC of STC rat interstitial, which promote intestinal peristalsis in slow transit constipation rats.