白术七物颗粒剂对气阴两虚型慢传输型便秘模型大鼠结肠组织c-kit的影响及其作用机制

Effect and Mechanism of Atractylodes Qiwu Granules on C-kit Expression Level in Colon Tissues of STC Rats with Qi-yin Deficiency

目的研究白术七物颗粒剂对气阴两虚型慢传输型便秘(STC)模型大鼠结肠组织c-kit的影响,探讨其可能作用机制。方法 2015年9—11月,将70只SPF级SD大鼠随机分成空白组20只、模型组50只,雌雄各半。模型组大鼠皮下注射盐酸吗啡注射液2.5 mg/kg,空白组注射等量0.9%氯化钠溶液,1次/d,连续注射45 d。模型组大鼠皮下注射盐酸吗啡注射液15 d后,灌服(青皮、枳壳、附子,剂量13 g/kg),连续30 d,建立气阴两虚型STC大鼠模型。模型建立成功后,将模型组剩下的40只大鼠随机分为模型对照组,白术七物颗粒高、低剂量组和莫沙必利组,每组10只,雌雄各半。除空白组、模型对照组外,白术七物颗粒高、低剂量组和莫沙必利组给予相应的药物灌胃。应用RT-PCR、免疫组化法检测白术七物颗粒剂对气阴两虚型STC模型大鼠结肠组织c-kit影响。结果模型组样本大鼠肠道推进率低于空白组样本(P<0.05)。白术七物颗粒低剂量组、白术七物颗粒高剂量组、莫沙必利组大鼠小肠推进率高于模型对照组(P<0.05)。白术七物颗粒高剂量组、莫沙必利组c-kit mRNA表达水平高于模型对照组(P<0.05)。白术七物颗粒低剂量组c-kit mRNA表达水平低于白术七物颗粒高剂量组(P<0.05)。白术七物颗粒高剂量组、莫沙必利组c-kit表达水平高于模型对照组(P<0.05)。结论白术七物颗粒剂治疗气阴两虚型STC模型大鼠的作用机制是通过增加结肠组织中c-kit表达水平来实现的。

Objective To investigate the changes of c-kit expression level in colon tissues of STC model rats with qi-yin deficiency under the intervention of Atractylodes qiwu granules, and to explore the possible mechanism of action of Atractylodes qiwu granules in treating the disease. Methods This study was conducted from September to November 2015. A total of 70 SPF grade SD rats were randomized into the blank group（ 10 males and 10 females） and the model group（ 25 males and 25 females）. Rats in the model group and blank group were subcutaneously injected 2. 5 ml/kg of morphine hydrochloride injection,2. 5 ml/kg of 0. 9% sodium chloride injection,once daily for 45 consecutive days,respectively. The rats in the model group were gavaged with 13 g/kg of the solution with dissolved granules of pericarpium citri reticulatae viride, fructus aurantii and radix aconite carmichaeli on the 16 th day of prescribing morphine hydrochloride injection,for 30 consecutive days to establish the STC rat models with qi-yin deficiency. The STC rat models with qi-yin deficiency were verified to be established successfully after the above 45 d-intervention by examining the sacrificed 10 of the rats from the model group. The rest 40 rats in the model group were randomized into the model control group, high-dose Atractylodes qiwu granules group, low-dose Atractylodes qiwu granules group,and mosapride group,with 10 evenly composed of males and females in each group. After stopping the previous intervention for 7 days,the model control group,high-dose and low-dose Atractylodes qiwu granules groups,mosapride group and blank group were gavaged with solution of 0. 9% sodium chloride injection,high-dose Atractylodes qiwu granules,low-dose Atractylodes qiwu granules,mosapride,0. 9% sodium chloride,respectively. RT-PCR and immunohistochemistry were used to detect the expression levels of c-kit in the colon tissue samples taken from the rats of all the groups. Results The intestinal propulsion rate of model group was lower than that of blank group（ P〈0. 05）. The model control group had lower intestinal propulsion rate than low-dose and high-dose Atractylodes qiwu granules groups and mosapride group（ P〈0. 05）.The expression levels of c-kit mRNA in the model control group were lower than those in low-dose and high-dose Atractylodes qiwu granules groups and mosapride group（ P〈0. 05）. The expression levels of c-kit mRNA in low-dose Atractylodes qiwu granules group were lower than those in high-dose Atractylodes qiwu granules group（ P〈0. 05）. The model control group had lower expression levels of c-kit than high-dose Atractylodes qiwu granules group and mosapride group（ P〈0. 05）.Conclusion Atractylodes qiwu granules acts by increasing the c-kit level in colon tissues to treat STC with qi-yin deficiency.