Exploration of the mechanism of Dahuang Sanwei Tablets on intestinal motility in constipated rats through SCF/c-Kit signaling pathway
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摘要: 目的 观察大黄三味片通过酪氨酸激酶生长因子受体(tyrosine kinase growth factor receptor，c-Kit)及其配体干细胞因子(stem cell factor，SCF)(SCF/c-Kit)信号通路对便秘大鼠肠道动力的影响。方法 将40只SD大鼠随机分为对照组、模型组、大黄三味片组、莫沙必利组、c-Kit抑制剂组，每组8只。模型组、大黄三味片组、莫沙必利组、c-Kit抑制剂组先给予盐酸洛哌丁胺8 mg/kg灌胃造模7 d，造模成功后大黄三味片组给予大黄三味片混悬液243 mg/(kg/d)灌胃治疗7 d，莫沙必利组大鼠继续给予1.6 mg/(kg/d)灌胃治疗7 d，c-Kit抑制剂组在给予大黄三味片混悬液灌胃治疗的同时给予10 mg/(kg/d)的c-Kit抑制剂Masitinib灌胃给药7 d。通过粪便性状、质地和含水率评价大黄三味片的治疗效果，测定玻璃小球排出时间、小肠炭末推进率和离体结肠纵行平滑肌肌条收缩力，评估大黄三味片对肠道动力的影响，计算各组大鼠结肠组织SCF、c-Kit mRNA和蛋白的表达情况。结果 与对照组比较，模型组大鼠的粪便含水率、玻璃小球排出时间、小肠炭末推进率、肌条收缩力和SCF、c-Kit的表达量显著下降(P＜0.05)；与模型组比较，大黄三味片组大鼠的粪便含水率、玻璃小球排出时间、小肠炭末推进率、肌条收缩力和SCF、c-Kit的表达量显著上升(P＜0.05)。结论 大黄三味片可通过调控SCF/c-Kit信号通路，提高便秘大鼠的粪便含水率，恢复便秘大鼠受损的结肠动力，对便秘大鼠具有显著的治疗效果。
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关键词:
- 大黄三味片 /
- 便秘 /
- 干细胞因子 /
- 酪氨酸激酶生长因子受体
Abstract: Objective To observe the effects of Dahuang Sanwei Tablets on intestinal secretion and gut microbiota in constipated rats.Methods Forty SD rats were randomly divided into normal control group(NC), model control group(MC), Dahuang Sanwei Tablets group(DS), mosapride group(MG) and c-Kit inhibitor group(CI), with 8 rats in each group. MC group, DS group, MG group and CI group were given loperamide hydrochloride 8 mg/kg, and after successful modeling, DS group was given 243 mg/(kg/d) of Dahuang Sanwei Tablets intragastric treatment for 7 days, MG group was given 1.6 mg/(kg/d) of mosapride intragastric treatment for 7 days, CI group was given 10 mg/(kg/d) of Masitinib intragastric treatment for 7 days. The therapeutic effect of Dahuang Sanwei Tablets was evaluated by tool character, texture and water content, and the intestinal motility was evaluated by measuring the time of exhalation of glass beads and the rate of intestinal carbon powder propulsive, the changes of the amplitude of the strips were observed and the contractility of the strips was compared. The expression of SCF and c-Kit mRNA and protein in the colonic tissues were calculated.Results Compared with the NC group, the water content of feces, the time of glass pellet excretion, the rate of intestinal carbon powder propulsive, the contractility of muscle strips and the expression of SCF and c-Kit in the model group were significantly decreased(P < 0.05), the water content of feces, the time of exhalation of glass beads, the rate of intestinal carbon powder propulsive, the contractility of muscle strips and the expression of SCF and c-Kit were significantly increased in DS group(P < 0.05).Conclusion Dahuang Sanwei Tablets have a significant therapeutic effect on rats with constipation. It can increase the water content of feces and restore the impaired colonic motility of rats with constipation through SCF/c-Kit signaling pathway. -
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表 1 本实验使用的引物序列
目的基因 上游引物序列(5'-3') 下游引物序列(5'-3') SCF TGACCTCGTGGCATGTATGG GGACTTTGCGGCTTTCCTATTAC c-Kit ATCAGGGCGACTTCAATTACG CTGCTGGTGTTCAGGTTTAGGG GAPDH CTGGAGAAACCTGCCAAGTATG GGTGGAAGAATGGGAGTTGCT 
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表 2 本实验使用的抗体及其稀释倍数
抗体名称 稀释倍数 生产厂家 货号 兔SCF抗体 1∶2 000 Peoteintech 26582-1-AP 兔c-Kit抗体 1∶1 000 Abcam ab256345 兔GAPDH抗体 1∶20 000 Peoteintech 10494-1-AP
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