基于气道黏液高分泌大鼠模型的苓桂术甘汤与肾气丸“同病异治”之作用环节研究

Comparative Study on Action of Linggui Zhugan Decoction and Shenqi Pill on Airway Mucus Hypersecretion in Rats

目的在前期已证实苓桂术甘汤、肾气丸可"同病异治"改善"短气,有微饮"的基础上,从作用环节角度,探讨《金匮要略》中"同病异治"的内涵。方法随机选取30只SD大鼠,采用丙烯醛雾化吸入方式复制气道黏液高分泌大鼠模型,另取10只大鼠为正常组。将模型大鼠随机分为3组,每组10只,分别为模型组、苓桂术甘汤组、肾气丸组。正常组与模型组给予纯净水灌胃,各给药组给予同体积的相应汤药,连续4周后,检测指标。ELISA法检测肺泡灌洗液中MUC5AC、MUC5B、IL-13、IL-1β、IL-8、TNF-α、NE、TGF-α、EGF浓度。结果与正常组比较,模型组大鼠肺泡灌洗液中黏蛋白MUC5AC、MUC5B和细胞因子IL-13、IL-1β、IL-8、TNF-α、NE、TGF-α、EGF的含量均升高(P <0. 05)。与模型组比较,苓桂术甘汤组与肾气丸组均表现出MUC5AC含量的降低(P <0. 05),而MUC5B含量无明显变化(P>0. 05);两给药组大鼠肺泡灌洗液中细胞因子的含量呈不同程度的下降,其中:苓桂术甘汤组和肾气丸组大鼠NE含量均下降(P <0. 05);苓桂术甘汤组大鼠EGF、IL-1β、TNF-α含量降低(P <0. 05),TGF-α含量无明显变化(P> 0. 05);肾气丸组大鼠TGF-α、IL-13、IL-8含量显著降低(P <0. 05),EGF含量无明显变化(P> 0. 05)。苓桂术甘汤组与肾气丸组之间比较,两给药组对MUC5AC的表达和NE、EGF、IL-1β、TNF-α含量的影响无明显差异(P> 0. 05);肾气丸组TGF-α、IL-13、IL-8含量显著降低(P <0. 05)。结论肾气丸和苓桂术甘汤均能改善气道黏液高分泌,即"同病-同证-异治"这一"同病异治"的解读的可能性是存在的,其内在机理之一可能体现在异方作用于疾病的不同环节。

Objective Preliminary research confirms that Linggui Zhugan Decoction and Shenqi Pill can relieve"shortness breath". And the difference between Linggui Zhugan Decoction group and Shenqi Pill had no significant. This study,based on the confirmed conclusion,discussed the connotation of"different treatments for the same disease"from the point of view of the action link. Methods Thirty rats were randomly selected as the model group,and the remaining 10 were treated as normal control group.Remodel acrolein-induced airway mucus hypersecretion in rats,rat model of airway mucus hypersecretion was established by inhalation of acrolein on 2 consecutive weeks. Rats were randomly divided into three groups: model group,Linggui Zhugan Decoction group and Shenqi Pill group,and 10 rats per group. Each group was given intragastric administration for 4 consecutive weeks from the third week. Normal group and model group were given intragastric administration by water. The test was conducted in 4 weeks. After 4 weeks,we observed the impact and difference of Linggui Zhugan Decoction and Shenqi Pillin on the key cytokines in the pathogenesis of airway mucus secretion. By enzyme-linked immunosorbent assay,we detected the rat mucins in bronchoalveolar lavage fluid of MUC5 AC,MUC5 B and cytokine IL-13,IL-1 beta,IL-8,TNF-α,NE,TGF – alpha and EGF. Results Compared with the normal group,the contents of mucin MUC5 AC,MUC5 B and cytokines IL-13,IL-1β,IL-8,TNF-α,NE,TGF-α and EGF were all increased in the model rat alveolar lavage( P < 0. 05). Compared with the model group,the content of MUC5 AC was decreased( P < 0. 05),while the content of MUC5 B was not significantly changed( P > 0. 05). The content of cytokines in the alveolar lavage solution of rats in the two groups was down-regulated. The NE content of the rats in Linggui Zhugan Decoction group and Shenqi Pill group was decreased( P < 0. 05). The contents of EGF,IL-1β and TNF-α were decreased in the rats of the Linggui Zhugan Decoction group( P < 0. 05),and there was no significant change in TGF-alpha content( P >0. 05). The contents of TGF-α,IL-13 and IL-8 in the rats of Shenqi Pill group were significantly reduced( P < 0. 05),and there was no significant change in EGF content( P > 0. 05). Compared the rats of Linggui Zhugan Decoction group with those in Shenqi Pill group,the expressions of MUC5 AC and the influence of NE,EGF,IL-1β and TNF-α were not significantly different between the two administration groups( P > 0. 05). The contents of TGF-α,IL-13 and IL-8 in Shenqi Pill group decreased significantly( P < 0. 05). Conclusion Linggui Zhugan Decoction and Shenqi Pill can both improve high airway mucus secretion by inhibiting the goblet cells metaplasia,sticky protein expression and inhibiting inflammatory reaction. Shenqi Pill can significantly reduce the expressions of IL-13 and IL-8 in BALF,and inhibit the NE/TGF-alpha/EGFR signaling pathway,etc. While Linggui Zhugan Decoction may reduce the expressions of IL-1 and TNF-α in BALF. That is "the same disease-the same syndrome-different prescriptions"may happen when different prescriptions act on different parts of the disease.