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目的:从肠易激综合征模型大鼠结肠黏膜蛋白激酶A(protein kinase A,PKA)、环腺苷酸(Cyclic adenosine monophosphate,cAMP)、P物质(substance P,SP)的表达变化探讨疏肝健脾方防治IBS的可能机制。方法:72只SPF级雄性SD大鼠按体质量随机分为正常组、模型组、治疗组(疏肝健脾方低、中、高剂量)和匹维溴胺对照组,每组12只。采用辣素+束缚应激复合法制备IBS动物模型。除正常组外,其余各组大鼠在造模成功后分别给予不同药物或生理盐水连续灌胃2周,末次给药后处死动物,截取结肠。采用HE染色法检测大鼠结肠病理组织学改变;醛品红-桔黄G法检测肥大细胞活性;Western-blot技术和RT-PCR法检测蛋白激酶A、环腺苷酸、P物质表达变化。结果:模型组大鼠肥大细胞阳性表达及数目明显增多,与治疗组比较有明显差异(P<0.05);模型组大鼠PKA、cAMP、SP基因和蛋白表达明显高于正常组(P<0.05);经中药治疗后各组基因及蛋白表达均有下调,其中以高剂量组下调明显PKA、cAMP、SP高剂量组基因表达分别为(1.24±0.09)、(1.11±0.05)、(1.00±0.06),PKA、cAMP、SP高剂量组蛋白表达分别为(0.43±0.08)、(0.25±0.07)、(0.25±0.03),差异有统计学意义(P<0.05)。低、中剂量组虽有下降趋势,但不具有统计学意义(P>0.05)。结论:疏肝健脾方可能通过调控依赖环腺苷酸的蛋白激酶A,下调P物质表达,从而降低内脏敏感性并改善胃肠动力,发挥治疗肠易激综合征的效应。
OBJECTIVE: To investigate the expression of protein kinase A (PKA), cyclic adenosine monophosphate (cAMP) and substance P (SP) in the intestinal mucosa of irritable bowel syndrome model rats The possible mechanism of Jianpi Fang in preventing and treating IBS. Methods: Seventy-two SPF male Sprague-Dawley rats were randomly divided into normal group, model group, treatment group (low, middle and high dose of Shuganjianpi Decoction) and piroxicam control group, 12 rats in each group. The animal model of IBS was prepared by the combination of the hot and the restraint stress. Except for the normal group, the other rats in each group were given different drugs or saline for 2 weeks after successful modeling. The animals were sacrificed after the last administration, and the colon was taken. The histopathological changes of colon in rats were detected by HE staining. The activity of mast cells was detected by aldehyde magenta-orange G method. The protein expression of protein kinase A, cyclic adenosine monophosphate and substance P was detected by Western-blot and RT-PCR. Results: The expression of PKA, cAMP, SP in model group was significantly higher than that in normal group (P <0.05), while the number and expression of mast cells in model group were significantly increased (1.24 ± 0.09), (1.11 ± 0.05) and (1.00 ± 0.05), respectively, in the high-dose PKA, cAMP and SP groups 0.06). The high-dose PKA, cAMP and SP groups were (0.43 ± 0.08), (0.25 ± 0.07) and (0.25 ± 0.03) respectively. The difference was statistically significant (P <0.05). Although the low and middle dose groups showed a downward trend, they were not statistically significant (P> 0.05). Conclusion: Shuganjianpi Recipe can down-regulate the expression of substance P by regulating protein kinase A dependent on cyclic adenosine monophosphate, thereby reducing visceral sensitivity and improving gastrointestinal motility and playing the role of treating irritable bowel syndrome.