六味安消胶囊联合莫沙必利治疗慢传输型便秘大鼠的疗效及机制

Study on clinical efficacy and mechanism of mosapride combined with Liuwei Anxiao capsule in the treatment of slow transit constipation of rats

目的探讨六味安消胶囊与莫沙必利联合使用治疗慢传输型便秘大鼠的疗效及其可能机制。方法共纳入130只无特定病原体级大鼠,均为成年健康雄性Wistar大鼠,平均体重(220.38±20.43) g。采用大黄灌胃法最终成功构造123只慢传输型便秘模型大鼠,并按照简单随机分组法分为3组,每组各41只。A组每日给予2 g/mL莫沙必利灌胃治疗,B组每日给予2 g/mL六味安消胶囊灌胃治疗,C组每日给予2 g/mL莫沙必利联合2 g/mL六味安消胶囊治疗,共灌胃14 d。评价3组大鼠的临床疗效、临床症状评分;蛋白质印迹法检测3组大鼠结肠组织中c-Kit、转化生长因子β1(TGF-β1)、Smad6/7蛋白的表达水平;免疫组织化学法检测结肠组织ICC细胞核内c-Kit的表达水平;酶联免疫吸附试验检测炎症因子[白细胞介素(IL)-1β、IL-6、IL-8、IL-10、C反应蛋白(CRP)、肿瘤坏死因子α(TNF-α)]的表达水平。结果 C组总有效率高于A组和B组(92.7%vs. 75.6%vs. 78.0%),且C组治疗后的临床症状评分明显高于A组和B组,差异均有统计学意义(P <0.05)。治疗后,C组结肠组织中c-Kit、Smad6/7蛋白的表达水平高于A组和B组,差异均有统计学意义(P <0.05),但3组TGF-β1蛋白的表达水平比较差异无统计学意义(P> 0.05)。阳性染色的c-Kit蛋白呈现棕黄色颗粒,主要分布在结肠ICC细胞核内;治疗后,C组c-Kit表达量高于A组和B组,差异有统计学意义(P <0.05)。治疗后,C组血清IL-1β、IL-6、IL-8、IL-10、CRP、TNF-α水平均低于A组和B组,差异均有统计学意义(P <0.05)。结论六味安消胶囊联合莫沙必利可改善慢传输型便秘大鼠的临床症状,其机制可能与调控GALNT1/TGF-β1信号通路,降低血清炎症因子水平或抑制ICC细胞转分化有关。

Objective To investigate the clinical efficacy and possible mechanism of mosapride combined with Liuwei Anxiao capsule in the treatment of slow transit constipation of rats. Methods A total of 130 SPF rats were included in this study,all of which were adult healthy male Wistar rats with an average body weight of( 220. 38 ± 20. 43) g. The rhubarb gavage method was used to successfully construct 123 slow transit constipation model rats and were included in the study. They were divided into three groups by a simple random grouping method,with 41 rats in each group. Group A was given 2 g/mL mosapride by gavage every day,group B was given 2 g/mL Liuwei Anxiao capsules every day,and group C was given 2 g/mL mosapride combined with 2 g/mL Liuwei Anxiao capsule every day treatment,gavage for 14 days. The clinical efficacy and clinical symptom scores of the three groups of rats were evaluated;the expression levels of c-Kit,transforming growth factor β1( TGF-β),and Smad6/7 protein in the colon tissue of the three groups of rats were detected by Western blotting;the expression of c-Kit in the ICC cell nucleus of the colon tissue levels were detected by immunohistochemistry;the expression levels of inflammatory factors [interleukin( IL)-1β,IL-6,IL-8,IL-10,C-reactive protein( CRP),tumor necrosis factor α( TNF-α) ] were detected by enzyme-linked immunosorbent assay.Results The total effective rate of group C was higher than that of group A and group B( 92. 7% vs. 75. 6% vs. 78. 0%),and the clinical symptom score of group C after treatment was significantly higher than that of group A and group B,the differences were statistically significant( P <0. 05). After treatment,the expression levels of c-Kit and Smad6/7 protein in the colon tissue of group C were higher than those of group A and group B,and the differences were statistically significant( P < 0. 05),but there was no significant difference in the expression levels of TGF-β1 protein in the three groups( P > 0. 05). The positively stained c-Kit protein showed brown particles,mainly distributed in the nucleus of colonic ICC cells,the expression of c-Kit in group C was higher than that in groups A and B,the difference was statistically significant( P < 0. 05).The levels of serum IL-1β,IL-6,IL-8,IL-10,CRP,and TNF-α in group C were lower than those in group A and group B,and the differences were statistically significant( P < 0. 05). Conclusion Mosapride combined with Liuwei Anxiao capsule can improve the clinical efficacy of slow transit constipation of rats,and its mechanism may be related to the regulation of the GALNT1/TGF-1 signaling pathway to reduce the level of serum inflammatory cytokines or inhibit ICC transdifferentiation.