从慢传输型便秘大鼠结肠c-kit/SCF的基因表达及信号传导通路探讨四磨汤的调节机制

Exploring Mechanism of Simon Decoction from Gene Expression and Signal Transduction Pathway of Colonic c-kit/SCF in Rats with Slow Transit Constipation

目的：研究四磨汤对慢传输结肠的c-kit/SCF的基因表达及信号传导通路影响,探索理气中药改善结肠动力的作用机制。方法：选取120只SD大鼠,随机分为正常组30只,造模组90只,造模组饲喂含复方苯乙哌啶的饲料,造模成功后,将90只大鼠再随机分为3组,一组为空白对照组,一组为盐水对照组（安慰剂组）,一组为四磨汤治疗组,每组30只。停药7 d后,分别从四组中各抽取10只大鼠,测首例黑便排出时间。结肠肌电检测,验证＂四磨汤＂对STC大鼠结肠肌电的影响效应。通过Realtime-PCR检测,分别检测c-kit和SCF的mRNA在STC结肠组织中的表达情况。计算机辅助图像分析表达情况。结果：四磨汤组大鼠首粒黑便排出时间较对照组和安慰剂组明显缩短（P〈0.05）;较正常组有所延长（P〉0.05）。四磨汤组大鼠肌电频率、振幅、频率变异系数、振幅变异系数与正常组相比无统计学意义（P〉0.05）,模型组大鼠肌电频率低于正常组（P〈0.05）,振幅、频率变异系数、振幅变异系数高于正常组,有统计学意义（P〈0.05）。c-kit mRNA相对含量四磨汤组高于对照组及安慰剂组（P〈0.05）,低于正常组（P〉0.05）。SCF mRNA相对含量四磨汤组高于对照组及安慰剂组（P〈0.05）,低于正常组（P〉0.05）。结论：STC模型大鼠的结肠c-kit和SCF基因表达下调,c-kit/SCF系统受损。四磨汤能够显著提高c-kit mRNA、SCF mRNA相对含量,显著抑制造模药物对大鼠结肠慢波的影响,是通过调节c-kit,SCF基因表达及相应的信号通路来治疗慢传输型便秘的。

Objective： To research the c-kit/SCF gene expression and signal transduction pathway of Simon Decoction on slow transit colon,exploring the mechanism of traditional Chinese medicine to improve the colonic motility of qi. Methods： A total of 120 SD rats were randomly divided into normal group（ n = 30） and model group（ n = 90）. The model used lomotil in feed. After the success of modeling,90 rats were randomly divided into 3 groups： the control group,saline control group（ placebo group）,Simon Decoction treatment group,30 rats in each group. 7 days after withdrawal,10 rats were selected from each of the four groups to measure the time of first black stool discharging. By Colonic electromyography detection,it verified the effect effect of Simon Decoction on colonic electromyography of STC rats. Through Realtime-PCR detection,we separately detected the expression of c-kit and SCF mRNA in STC colon tissue. Computer aided the image analysis. Results： The first time of defecation of rats in the Simon Decoction treatment group was significantly shorter than that of the control group and the placebo group（ P 0. 05） and was increased compared with the control group（ P 0. 05）. In the Simon Decoction treatment group,the frequency,amplitude and frequency of EMG amplitude variation coefficient and coefficient of variation had no statistical significance compared with those of the normal group（ P 0. 05）. EMG frequency of rats in model group was lower than that of the normal group（ P 0. 05）. The amplitude,frequency,amplitude variation coefficient and variation coefficient were higher than thsoe of the normal group,with statistical significance（ P 0. 05）. The relative content of c-kit and SCF mRNA of the Simon Decoction treatment group was more than that of the control group and the placebo group（ P 0. 05） and lower than that of the normal group（ P 0. 05）. Conclusion： STC model rats＇ expressions of c-kit and SCF in colon was down regulated and the c-kit/SCFsystem was damaged. Simon Decoction can improve c-kit mRNA and SCF mRNA relative content and inhibit the modeling of drug effects on rat colonic slow wave through regulating c-kit,expression of SCF gene and its signal transduction pathway to treat STC.