“从小便去之”对气道黏液高分泌模型大鼠肾素-血管紧张素-醛固酮系统和细胞因子的影响研究

Removing the Pathogenic Factor from Urine on RASS System and Cytokines in a Rat Model of Acrolein-induced Airway Mucus Hypersecretion

背景《金匮要略》一书中共计5处运用肾气丸,其主治与小便有关,主要功效为温补肾气,化气行水。目的以"夫短气有微饮,当从小便去之,苓桂术甘汤主之,肾气丸亦主之"为例,观察肾气丸对气道黏液高分泌模型大鼠肾素-血管紧张素-醛固酮(RASS)系统和细胞因子的影响。方法 2017年4-8月,采用随机数字表法将60只SPF级雄性SD大鼠分为正常组、模型组、肾气丸组(温肾利水)、苓桂组(化气利水)、肾气丸去苓桂组(温养肾气),每组12只。模型组、肾气丸组、苓桂组、肾气丸去苓桂组大鼠雾化吸入3 mg/L丙烯醛,3 h/次,2次/d,持续4周,进行造模;正常组雾化吸入等量0.9%氯化钠溶液。干预第5周开始,肾气丸组给予肾气丸药液灌胃,苓桂组给予苓桂汤灌胃,肾气丸去苓桂组给予肾气丸去苓桂汤灌胃,均1 ml/100 g、2次/d、持续4周;正常组和模型组给予等量蒸馏水灌胃。统计各组大鼠24 h尿量、肺功能参数[第0.3秒用力呼气末容积(FEV0.3)、用力肺活量(FVC)、第0.3秒用力呼气末容积占用力肺活量百分比(FEV0.3/FVC%)、最大呼气流量(PEF)]、肺泡灌洗液(BALF)中白介素(IL)-1β、IL-13、转化生长因子(TGF)-α、肿瘤坏死因子(TNF)-α和血清中外周血肾素活性(PRA)、血管紧张素Ⅱ(AngⅡ)、醛固酮(ALD)。结果模型组、肾气丸组、苓桂组、肾气丸去苓桂组大鼠24 h尿量高于正常组(P<0.05);肾气丸组、苓桂组、肾气丸去苓桂组大鼠24 h尿量高于模型组(P<0.05);苓桂组大鼠24 h尿量高于肾气丸组(P<0.05)。模型组大鼠FEV0.3、FVC、FEV0.3/FVC%、PEF低于正常组(P<0.05);苓桂组大鼠FEV0.3/FVC%低于正常组(P<0.05);肾气丸组、苓桂组、肾气丸去苓桂组大鼠FEV0.3、FVC、PEF高于模型组(P<0.05);肾气丸组、肾气丸去苓桂组大鼠FEV0.3/FVC%高于模型组(P<0.05)。模型组、苓桂组、肾气丸去苓桂组大鼠BALF中IL-1β、IL-13、TGF-α、TNF-α高于正常组(P<0.05);肾气丸组大鼠BALF中TNF-α高于正常组(P<0.05);肾气丸组、苓桂组、肾气丸去苓桂组大鼠BALF中IL-1β、IL-13、TGF-α、TNF-α低于模型组(P<0.05);苓桂组、肾气丸去苓桂组大鼠BALF中IL-1β、TNF-α高于肾气丸组(P<0.05)。各组大鼠血清中PRA比较,差异无统计学意义(P>0.05);模型组大鼠血清中AngⅡ、ALD高于正常组(P<0.05);苓桂组、肾气丸去苓桂组大鼠血清中ALD高于正常组(P<0.05);肾气丸组、苓桂组、肾气丸去苓桂组大鼠血清中AngⅡ、ALD低于模型组(P<0.05);肾气丸去苓桂组大鼠血清中ALD高于肾气丸组(P<0.05)。结论肾气丸在调控气道黏液高分泌大鼠BALF中IL-1β、TGF-α方面优于苓桂、肾气丸去苓桂;温养肾气、化气利水皆对气道黏液高分泌大鼠具有一定的干预作用,二者合用则效果更佳,即所谓温养肾气和化气利水之药合用则旧饮易去而新饮不留。肾气丸"从小便去之"改善气道黏液高分泌模型大鼠症状可能与改善其肺功能、抑制RASS系统被激活、调节黏液高分泌相关细胞因子有关。

Background In Synopsis of Prescriptions of the Golden Chamber,Shenqi pills are used in a total of 5 prescriptions,mainly for the treatment of urination problems via warming and nourishing the kidney qi,circulating qi and promoting urination.Objective To observe the effect of Shenqi pills on renin-angiotensin-aldosterone(RASS)system and cytokines in a rat model of acrolein-induced airway mucus hypersecretion based on the theory of "Treatment of shortness of breath caused by minor case of Fluid-retention syndrome should following the principle of removing pathogenic factors from urine.Linggui Zhugan Decoction and Shenqi pills can be adopted" proposed in Synopsis of Prescriptions of the Golden Chamber.Methods This study was carried out between April and August 2017.A total of 60 SPF male Sprague-Dawley rats were randomly selected and equally divided into control group,model group,Shenqi pills group,Linggui(poria cocos and cassia twig)group and Shenqi pills without Linggui group.The latter 4 groups were given atomization inhalation of 3 mg/L acrolein,twice daily at an interval of 3 hours for 4 consecutive weeks for establishing the models of acrolein-induced airway mucus hypersecretion,while the control group received atomization inhalation of the same dose of 0.9% sodium chloride solution.From the 5 th week of the intervention,Shenqi pills group,Linggui group,and Shenqi pills without Linggui group received 1 ml/100 g Shenqi pills solution for warming the kidney qi and promoting urination,1 ml/100 g Linggui decoction for circulating qi and promoting urination,1 ml/100 g Shenqi pills without Linggui decoction for warming and nourishing the kidney qi,respectively,twice daily for another 4 consecutive weeks,while control and model groups received intragastric administration of the same volume of distilled water.24-hour urine volume,pulmonary function parameters(FEV0.3,FVC,FEV0.3/FVC,PEF),levels of IL-1β,IL-13,TGF-αand TNF-αin bronchoalveolar lavage fluid(BALF),and levels of PRA,plasma angiotensin Ⅱ and aldosterone(ALD)were collected.Results 24-hour urine volume in the control group was less than that of other groups(P<0.05).Model group showed less 24-hour urine volume than 3 medicine intervened model groups(P<0.05).Shenqi pills group had less 24-hour urine volume than Linggui group(P<0.05).Compared with the control group,model group demonstrated lower average FEV0.3,FVC,FEV0.3/FVC and PEF,and Linggui group showed less average FEV0.3/FVC(P<0.05).Model group demonstrated lower FEV0.3,FVC,and PEF than Linggui group,and showed lower FEV0.3,FVC,PEF and FEV0.3/FVC ratio than both Shenqi pills and Shenqi pills without Linggui group(P<0.05).The control group showed lower IL-1β,IL-13,TGF-α and TNF-α in BALF compared with other groups(P<0.05).The Shenqi pills group showed higher levels of TNF-αin BALF than control group(P<0.05).And the levels of these 4 parameters were lower in 3 medicine-intervened model groups than those of the model group(P<0.05).Moreover,Shenqi pills group showed lower levels of IL-1β and TNF-α in BALF than both Linggui and Shenqi pills without Linggui groups(P<0.05).PRA levels in all groups were similar(P>0.05).Compared with the control group,model group showed higher plasma angiotensin Ⅱ and ALD levels(P<0.05),and Linggui and Shenqi pills without Linggui groups showed higher plasma ALD levels(P<0.05).Model group demonstrated higher plasma angiotensin Ⅱ and ALD levels than 3 medicine intervened model groups(P<0.05).Shenqi pills without Linggui group had higher plasma ALD compared with Shenqi pills group(P<0.05).Conclusion Shenqi pills showed better effect than Linggui and Shenqi pills without Linggui in regulating IL-1β and TGF-α in BALF.Fluid-retention syndrome can be alleviated by warming and nourishing kidney qi or circulating qi and promoting urination,but the combination of the two can yield better efficacy,namely,the retained excessive old fluid and the new-produced excessive fluid can be removed.The mechanism of removing the pathogenic factor from urine might be related to improve pulmonary functions,inhibit RASS system activation and regulate cytokines associated with mucus hypersecretion.