大黄素止咳祛痰作用的实验研究

Experimental study on antitussive and expectorant effects of Emodin

目的探究中药单体大黄素的止咳祛痰作用。方法本研究为实验研究,采用随机数字表法分为模型组(5只,生理盐水灌胃)、磷酸可待因治疗组(6只,0.03 g·kg^(-1)·d^(-1))、大黄素止咳组(5只,0.2 g·kg^(-1)·d^(-1)),用浓氨水处理小鼠,制备咳嗽模型,观察大黄素对小鼠的止咳作用;通过酶联免疫吸附试验检测空白对照组(5只,不吸入浓氨水,不灌胃)及浓氨水引咳实验中模型组和大黄素止咳组小鼠血清中白细胞介素6水平。将小鼠分为阴性对照组(5只,生理盐水灌胃)、氯化铵治疗组(6只,1 g·kg^(-1)·d^(-1))、大黄素祛痰组(5只,0.2 g·kg^(-1)·d^(-1)),采用气管酚红法和肺组织糖原染色评估大黄素的祛痰作用,通过实时荧光定量聚合酶链反应法检测阴性对照组、氯化铵治疗组、大黄素祛痰组小鼠肺组织中黏蛋白5AC(MUC5AC)mRNA的表达水平。结果使用浓氨水处理模型组、磷酸可待因治疗组和大黄素止咳组小鼠,与模型组相比,大黄素可延长小鼠咳嗽潜伏期[(1.8±0.8)s比(53.2±1.9)s,P<0.01],并减少小鼠咳嗽次数[(115.4±32.3)次/3 min比(14.0±1.7)次/3 min,P<0.001],降低血清中白细胞介素6水平[(551.33±62.52)ng/L比(208.24±8.77)ng/L,P<0.001]。通过小鼠气管酚红法评估阴性对照组、氯化铵治疗组和大黄素祛痰组小鼠的气道排痰能力发现,与阴性对照组相比,大黄素可增加小鼠气管酚红的排泄量[(0.88±0.14)mg/L比(7.54±1.71)mg/L,P<0.001],降低小鼠肺组织中MUC5AC mRNA表达水平[(1.00±0.13)比(0.31±0.03),P<0.001]。通过糖原染色观察阴性对照组、氯化铵治疗组和大黄素祛痰组小鼠的肺组织发现,大黄素能减少小鼠肺组织中杯状细胞的数量。结论大黄素有一定的止咳祛痰的作用。

Objective To explore the antitussive and expectorant effects of Emodin.Methods This was an experimental study.Using random number table,mice were divided into model group(n=5,gavage of normal saline),codeine phosphate group(n=6,0.03 g·kg^(-1)·d^(-1)),and Emodin cough relieving group(n=5,0.2 g·kg^(-1)·d^(-1)).The mice were treated with concentrated ammonia water to prepare a cough model,and the anticough effect of Emodin on mice was observed.The serum levels of interleukin-6 were detected by enzyme-linked immunosorbent assay in the blank control group(n=5,no inhaling concentrated ammonia water,no instillation),the cough model group induced by concentrated ammonia water,and the Emodin cough relieving group.Mice were divided into negative control group(n=5,normal saline administration),ammonium chloride treatment group(n=6,1 g·kg^(-1)·d^(-1)),and Emodin expectorant group(n=5,0.2 g·kg^(-1)·d^(-1)).Mouse trachea phenol red method and mouse lung glycogen staining were used to evaluate the expectorant effect of Emodin.Quantitative real-time PCR was used to detect recombinant mucin 5 subtype AC(MUC5AC)mRNA expression in lung tissues of mice in negative control group,ammonium chloride treatment group,and Emodin expectorant group.Results The mice in the model group,the codeine phosphate treatment group,and the Emodin cough relief group were treated with concentrated ammonia water.Compared with the model group,Emodin could significantly prolong the cough latency of mice([1.8±0.8]s vs[53.2±1.9]s,P<0.001),reduce the cough frequency of mice([115.4±32.3]/3 mins vs[14±1.7]/3 mins,P<0.001),and decrease the expression level of interleukin-6 in serum([551.33±62.52]ng/L vs[208.24±8.77]ng/L,P<0.001).Airway sputum excretion ability of mice in the negative control group,ammonium chloride treatment group,and Emodin expectorant group was evaluated by tracheal phenol red method.Compared with the negative control group,Emodin increased tracheal phenol red excretion([0.88±0.14]mg/L vs[7.54±1.71]mg/L,P<0.001),and decreased the expression level of MUC5AC mRNA in lung tissue([1.00±0.13]vs[0.31±0.03],P<0.001).The lung tissues of mice in negative control group,ammonium chloride treatment group and Emodin expectorant group were observed by glycogen staining,and it was found that Emodin could reduce the number of goblet cells in the lung tissue of mice.Conclusions Emodin has certain antitussive and expectorant effects.