疏肝健脾方调控PKA-TRPV1-CGRP通路治疗腹泻型肠易激综合征的临床和实验研究

发布时间：2018-04-27 18:41

本文选题：腹泻型肠易激综合征 + 疏肝健脾方　；参考：《广西中医药大学》2017年硕士论文

【摘要】：第一部分临床研究目的:运用疏肝健脾方干预肝郁脾虚证腹泻型肠易激综合征,观察治干预前后腹痛、腹泻等症状的变化情况,评价疏肝健脾方治疗肝郁脾虚证腹泻型肠易激综合征的临床疗效。方法:选择符合纳入标准的肝郁脾虚证腹泻型肠易激综合征患者48例,随机分为疏肝健脾方治疗组24例、西药匹维溴铵对照组24例。治疗组采用疏肝健脾方(柴胡12 g、白芍20 g、陈皮10 g、炒白术15 g、防风10 g、炮姜5 g、炙甘草10 g)干预,选用农本方制剂,每日1剂,200ml,2次/日,冲服。对照组口服匹维溴铵,每次0.2 g,一天三次。两组均以连续用药4周为一个疗程。比较两组干预前后症状总积分、各临床症状积分及干预后总疗效,作出综合评定。结果:总体临床疗效方面:治疗组24例中,临床痊愈5例,显效12例,有效4例,无效3例,总有效率为87.5%;对照组24例中,临床痊愈6例,显效9例,有效5例,无效4例,总有效率为83.3%,两组的总体临床疗效相当(P0.05)。两组干预前后总积分比较均有显著性差异(P0.05),提示治疗组与对照组对改善临床症状均有明显作用。干预后治疗组与对照组比较症状总积分有显著性差异,说明治疗组对临床症状的改善优于对照组。治疗组对缓解腹痛程度、腹痛频率的疗效优于对照组(P0.05);对改善腹泻及大便形状与对照组的疗效相当(P0.05)。结论:疏肝健脾方治疗肝郁脾虚证IBS-D能改善其临床症状,从临床角度验证了疏肝健脾方是治疗肝郁脾虚证IBS-D的有效方剂。第二部分实验研究目的:观察疏肝健脾方对大鼠结肠黏膜肥大细胞及PKA、TRPV1、CGRP表达的影响。方法:将48只雄性SD大鼠按体重随机分为模型组、正常组、低剂量组、中剂量组、高剂量组和对照组。正常组除外,其余各组均采用束缚应激结合辣素灌胃,按照药物/体重0.5mg/kg的剂量灌胃,2次/周,连续2周,制备IBS模型。从第3周末起,低剂量组、中剂量组及高剂量组分别按3.1g/kg·d,6.2g/kg·d,12.4g/kg·d剂量给予疏肝健脾方中药;对照组予匹维溴铵水溶液15mg/kg·d;模型组予等剂量生理盐水,正常组予同步饲养。大鼠灌胃给药体积为4ml/100g,连续灌胃2周。第5周末,处死所有大鼠,截取回盲瓣下2cm至肛门上1.5cm的全部结肠,行HE染色及肥大细胞染色,进行相关病理检测;运用于荧光定量PCR和Western-blot技术检测PKA、TRPV1及CGRP的表达。结果:模型组肥大细胞表达比正常组明显增多,差异有统计学意义(P0.05)。低剂量组、中剂量组、高剂量组肥大细胞呈逐渐减少趋势,与模型组比较,差异有统计学意义(P0.05)。说明IBS-D大鼠存在肥大细胞高表达水平,疏肝健脾方具有稳定或降低肥大细胞表达的效用。荧光定量PCR和Western-blot技术检测结果显示,模型组大鼠结肠组织PKA、TRPV1、CGRP基因和蛋白表达明显高于正常组,差异均有统计学意义(P0.05)。干预后,与模型组相比,高剂量组、对照组PKA、TRPV1、CGRP基因和蛋白表达以及中剂量组CGRP基因表达均呈下降趋势,差异有统计学意义(P0.05)。而低剂量、中剂量组PKA、TRPV1及低剂量组CGRP基因表达与模型组比较,差异均无统计学意义(P0.05)。低剂量、中剂量组PKA、TRPV1及CGRP蛋白表达与模型组比较,差异均无统计学意义(P0.05)。结论:疏肝健脾方治疗腹泻型肠易激综合征的机制可能与下调PKA表达,介导TRPV1敏化,减少CGRP释放有关。
[Abstract]:The first part of clinical study objective: To observe the diarrhea type irritable bowel syndrome of liver qi stagnation and spleen deficiency syndrome, observe the changes of abdominal pain, diarrhea and other symptoms before and after treatment, and evaluate the clinical effect of the treatment of diarrhea type irritable bowel syndrome of stagnation of liver qi and spleen deficiency syndrome. Methods: choose the liver qi stagnation and spleen deficiency syndrome in accordance with the standard. 48 patients with diarrhea irritable bowel syndrome were randomly divided into 24 cases of Shugan Jianpi Fang treatment group and 24 cases of Western medicine pippium bromide control group. The treatment group was treated with Shugan Jianpi Recipe (Radix Bupleuri 12 g, Radix Paeoniae Radix Paeoniae 20 g, orange peel 10 g, stir frying Baizhu 15 g, anti wind 10 g, ginger 5 g, Radix Glycyrrhiza 10 g) and selected agricultural herbs, 1, 200ml, 2 times / day, and control group. 0.2 g each time, three times a day. Two groups were treated with continuous medication for 4 weeks. The total score of symptoms before and after the intervention of the two groups, the integral of clinical symptoms and the total effect after intervention were evaluated comprehensively. Results: in the total clinical efficacy, 5 cases were cured in 24 cases, 12 cases were cured, 4 cases were effective, 3 cases were invalid and total effective. Among the 24 cases in the control group, 6 cases were cured, 9 cases were markedly effective, 5 cases were effective and 4 cases were invalid, the total effective rate was 83.3%, the total clinical effect of two groups was equal (P0.05). The total integral before and after the intervention of the two groups had significant difference (P0.05), suggesting that the treatment group and the control group had obvious effect on the improvement of clinical symptoms. The treatment group and the control group had a significant effect on the clinical symptoms. The treatment group and the control group were treated with the control group and the control. The curative effect of the treatment group was better than the control group. The curative effect of the treatment group on the degree of abdominal pain and the frequency of abdominal pain was better than that of the control group (P0.05); the effect of improving the shape of diarrhea and stool with the control group was equal (P0.05). Conclusion: the IBS-D of the liver qi and spleen deficiency prescription in the treatment of liver depression and spleen deficiency can improve it. Clinical symptoms, from the clinical point of view that the Shugan Jianpi prescription is an effective prescription for the treatment of liver depression and spleen deficiency syndrome IBS-D. The second part of the experimental study: To observe the effect of Shugan Jianpi Recipe on the expression of mast cells and PKA, TRPV1, and CGRP in the colon mucosa of rats. Methods: 48 male SD rats were randomly divided into model group, normal group, low dose group, In the middle dose group, the high dose group and the control group, except the normal group, the other groups were combined with the binding stress combined with the gavage of the stomach. The dose of 0.5mg/kg was given to the stomach, 2 times per week for 2 weeks, and the IBS model was prepared. From the third weekend, the low dose group, the middle dose group and the high dose group were given 3.1g/kg. D, 6.2g/kg. D, 12.4g/kg. D dosage, respectively. The control group was treated with 15mg/kg D in the control group. The model group was given equal dose of normal saline, and the normal group was fed with the normal group. The rats were given 4ml/100g for 2 weeks. The rats were sacrificed at the fifth weekend. All the rats were killed and the whole colon of 2cm to 1.5cm on the anus was intercepted, and HE staining and mast cell staining were performed. The expression of PKA, TRPV1 and CGRP was detected by fluorescence quantitative PCR and Western-blot. Results: the expression of mast cells in the model group was significantly higher than that in the normal group, and the difference was statistically significant (P0.05). The mast cells in the low dose group, middle dose group and high dose group decreased gradually, and the difference was statistically significant compared with the model group. Meaning (P0.05). It shows that the hyperexpression level of mast cells in IBS-D rats is high, and the prescription of soothing the liver and invigorating spleen can stabilize or reduce the expression of mast cells. The results of fluorescence quantitative PCR and Western-blot techniques show that the expression of PKA, TRPV1, CGRP gene and protein in the colon tissue of the model group is significantly higher than that in the normal group, the difference is statistically significant (P0.05). Compared with the model group, the PKA, TRPV1, CGRP gene and protein expression of the control group and the expression of CGRP gene in the middle dose group showed a decreasing trend compared with the model group, and the difference was statistically significant (P0.05). The low dose group, PKA, TRPV1 and low dose group CGRP gene tables were compared with the model group, and the difference was not statistically significant (P0.05). The expression of PKA, TRPV1 and CGRP protein in the medium dose group was compared with the model group, and the difference was not statistically significant (P0.05). Conclusion: the mechanism of the treatment of diarrhea type irritable bowel syndrome by Shugan Jianpi Recipe may be related to the downregulation of PKA expression, the mediating TRPV1 sensitization and reducing the release of CGRP.

【学位授予单位】：广西中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R259

【参考文献】