金水六君煎拮抗地塞米松治疗慢性阻塞性肺病大鼠阴虚痰饮的实验研究

Experimental study of Jinshuiliujunjian on the treatment of COPD and its antagonism against dexamethasone

目的:探讨金水六君煎治疗慢性阻塞性肺疾病(COPD)阴虚痰饮的作用和机制。方法:SD大鼠60只,随机分成空白组、模型组、地塞米松0.5 mg/kg组、地塞米松+金水六君煎2.62、5.25、10.5 g/kg组。采用烟熏联合气道脂多糖滴入建立COPD大鼠模型,地塞米松肌注诱导大鼠阴虚状态。观察大鼠一般状态、肺功能、酚红排泌量,PAS染色法测定气道杯状细胞数,RT-PCR、Western法检测肺组织黏蛋白5AC(MUC5AC)和水通道蛋白5(AQP5)基因与蛋白表达,ELISA法测定血清ACTH、CORT、cAMP、cGMP、cAMP/cGMP含量。结果:模型组大鼠肺功能明显下降,气道杯状细胞数、酚红排泌量及MUC5AC表达明显增多,AQP5表达明显下降。地塞米松组大鼠出现形体消瘦,饮水量增多,毛发干枯,易激惹等阴虚表现,血清ACTH、CORT、cAMP、cGMP降低,cAMP/cGMP升高。地塞米松组及地塞米松+金水六君煎各组大鼠肺功能明显改善,气道杯状细胞数、酚红排泌量及MUC5AC表达明显减少。地塞米松+金水六君煎各组大鼠阴虚症状较地塞米松明显减轻,血清ACTH、CORT接近正常对照水平,cAMP、cAMP/cGMP明显降低,肺组织AQP5表达显著上调。结论:金水六君煎养阴作用可能与拮抗糖皮质激素对肾上腺皮质抑制,纠正环核苷酸代谢失调有关。化痰作用可能与促进AQP5表达相关。

Objective: To explore the effect of Jinshuiliujunjian on COPD and its Yinxutanyin mechanism. Methods: 60 SD rats were randomly divided into the control group,the model group, 0.5 mg/kg dexamethasone group, dexamethasone+ Jinshuiliujunjian combination groups(2.62,5.25,10.5 g/kg). COPD model was established by cigarette smoking combined with LPS instillation in airway. Yinxu symptoms of rats was established by intramuscular injection of dexamethasone. Rats general state, lung function and phenol red excretion were observed. PAS staining was used to determine the number of goblet cells and mucus in airway. qRT-PCR and Western blot were applied to verify the mRNA and protein expression levels of MUC5AC and AQP5. Serum contents of ACTH, CORT, cAMP, cGMP and cAMP/cGMP were detected by ELISA. Results: In the model group, the lung function and expression of AQP5 were significantly decreased, and the number of goblet cells, phenol red excretion and the expression of MUC5AC were significantly increased. In dexamethasone group, rats showed Yinxu symptoms as weight loss, increased water intake, dry hair and irritability. The levels of ACTH, CORT, cAMP and cGMP were significantly decreased, and cAMP/cGMP was significantly increased in dexamethasone group. In dexamethasone group and dexamethasone+ Jinshuiliujunjian combination groups, lung function, reduced airway goblet cells, phenol red excretion and the expression of MUC5AC were significantly improved. Yinxu symptoms in dexamethasone+ Jinshuiliujunjian combination groups were alleviated. Levels of ACTH and CORT were close to normal levels, levels of cAMP and cAMP/cGMP were significantly decreased, and the expression of AQP5 was significantly increased. Conclusion: Jinshuiliujunjian has the effect on Yinxu symptoms, its mechanism may be related to antagonism against glucocorticoid inhibiting the adrenal cortex, and regulating the cyclic nucleotide metabolism disorder. Its Huatan function may be related to promoting the expression of AQP5.