基于肺水肿模型探讨葶苈子升降浮沉药性

Ascending,Descending,Floating and Sinking Properties of Descurainiae Semen Lepidii Semen Based on Pulmonary Edema Model

目的:基于葶苈子药性沉降,根据“病位在里者宜沉降”的治则,建立病位在里的肺水肿模型,结合病势趋向的改变,验证葶苈子药性“沉降”的科学性,以期初步阐释中药升降浮沉药性的科学内涵。方法:选择60只雄性SD大鼠,随机分为假手术组、模型组(20 mg·kg^(-1))、阳性药地塞米松组(0.075 mg·kg^(-1))、葶苈子低、中、高剂量(1.167、2.334、4.668 g·kg^(-1))组并通过胸膜腔注射1%角叉菜胶(2 mL·kg^(-1))建立肺水肿模型,检测肺水肿评价指标(肺剖检、胸腔渗出液量、白细胞数量、肺湿重/干重比、肺含水量及肺通透性),确定葶苈子干预肺水肿的最佳剂量;检测与机体气机调节密切相关的五大系统(中枢系统、呼吸系统、循环系统、消化系统、泌尿系统)相关指标,观察葶苈子干预对肺水肿大鼠病势趋向的改变,确定其升降浮沉之性;并观察各组大鼠苏木素-伊红(HE)染色切片、炎症细胞种类和数目等,初步探究葶苈子改善肺水肿的作用机制。结果:与假手术组比较,胸腔积水量和胸腔积水中白细胞渗出量显著增加(P<0.01),肺湿重/干重比、肺含水量及肺通透性显著升高(P<0.01),并出现咳嗽、喘促、呼吸困难、弓背现象,少量大鼠鼻子湿润,鼻孔出现泡沫状液体等症状,剖检时肺出现体积增大或伴有瘀血,气管处出现大量粉红色泡沫状液体;与模型组比较,葶苈子可减少大鼠胸腔积水量和胸腔积水中白细胞渗出量,降低肺脏器系数、肺湿重/干重及肺含水量,改善肺组织水肿出血等,且以葶苈子中剂量治疗肺水肿效果最佳(P<0.01);对于呼吸系统,与假手术组比较,模型组大鼠咳嗽潜伏时间、引喘潜伏时间明显减少(P<0.05,P<0.01),咳嗽次数和喘息次数显著增加(P<0.01),与模型组比较,葶苈子低、中、高剂量组均显著增加咳嗽潜伏期、引喘潜伏期,减少咳嗽次数、喘息次数(P<0.01);对于泌尿系统,与假手术组比较,模型组大鼠显著减少尿量,葶苈子中、高剂量显著增加尿量(P<0.01),低剂量组明显增加尿量(P<0.05),但均对排汗无影响;对于消化系统,与假手术组比较,模型组大鼠胃残留率显著增加(P<0.01),胃排空率、小肠推动率显著下降(P<0.01),胃泌素(GT)明显增加(P<0.05),和模型组比较,葶苈子低剂量组显著增加小肠推动率(P<0.01),中、高剂量组可显著增加胃排空率、小肠推动率(P<0.01),显著减少胃残留率(P<0.01),显著或显著减少GT以促进胃肠运动及胃肠道的运输(P<0.01),增加胃动素(MTL)促进大鼠的胃排空(P<0.05,P<0.01);对于循环系统,与假手术组比较,大鼠左室射血分数(LVEF)、左室短轴缩短率(LVFS)、心输出量(CO)显著降低(P<0.01),模型组大鼠的心率有升高趋势,收缩压(SBP)显著升高(P<0.01),舒张压(DBP)明显升高(P<0.05);与模型组比较,葶苈子低剂量组明显增加LVEF、降低大鼠的DBP(P<0.05),葶苈子中剂量组显著增加LVEF、LVFS、CO和SBP(P<0.01),明显降低大鼠的DBP(P<0.05),葶苈子高剂量组显著增加LVFS(P<0.01),显著降低大鼠的SBP(P<0.01),明显降低大鼠的DBP(P<0.05);对于中枢系统,与假手术组比较,模型组大鼠站立次数显著降低(P<0.01),葶苈子明显减少大鼠的运动距离、运动时间、站立次数和在旷场中心活动时间,增加静止时间和旷场边缘活动时间(P<0.05,P<0.01);此外,与假手术组比较,模型组大鼠肺管腔周围的炎症浸润严重,气管增厚、内有水肿液聚集,肺组织破坏严重,血中白细胞计数、中性粒细胞比例显著增加(P<0.01),单核细胞比例明显增加(P<0.05),肺泡灌洗液中γ干扰素(IFN-γ)显著降低(P<0.01),白细胞介素-4(IL-4)明显提高(P<0.05)、免疫球蛋白E(IgE)水平显著升高(P<0.01),肺组织活性氧(ROS)水平显著升高(P<0.01),与模型组比较,葶苈子能减少白细胞计数和中性粒细胞积聚,减少肺小血管充血和肺间质水肿,降低肺泡灌洗液中IFN-γ和IL-4水平,升高IgE水平,降低肺组织ROS水平(P<0.05,P<0.01)。结论:葶苈子对病位在里的肺水肿模型具有显著改善作用,通过泻水逐饮、调节水液排泄,降泻肺气、调节气机、泻肺气之壅闭,促肺气肃降、调节气机下行,提示葶苈子作用趋势为沉降。其中以葶苈子中剂量作用最佳,其作用机制可能是通过调控中性粒细胞炎症反应来发挥干预作用的。

Objective:The pulmonary edema(PE)model where the disease was located in the viscera was established according to the treatment principle that patients with the disease location inside should be treated with descending and sinking medicine,combined with changes in the disease tendency,to verify the scientificity of descending and sinking properties of Descurainiae Semen Lepidii Semen(SD),and to preliminarily elucidate the scientific connotation of descending,ascending,floating and sinking of Chinese medicine.Method:Sixty male SD rats were randomly divided into normal control group,model group(20 mg·kg^(-1)),positive drug group(dexamethasone,0.075 mg·kg^(-1))and SD low(1.167 g·kg^(-1)),medium(2.334 g·kg^(-1))and high(4.668 g·kg^(-1))dose groups.The PE model was established by intrapleural injection of1% carrageenan(2 mL·kg^(-1)).The evaluation indexes(lung autopsy,amount of pleural effusion,number of white blood cells,lung wet/dry weight ratio,lung water content and lung permeability)were tested to determine the optimal dose of SD decoction for intervention of PE.The relevant indexes of the five major systems(central nervous system,respiratory system,circulatory system,digestive system and urinary system)closely related to the body’s Qi movement were detected and changes in the disease tendency in PE rats were analyzed,to determine the descending,ascending,floating and sinking properties of SD.In addition,histopathological changes were investigated by hematoxylin-eosin(HE)staining,and types and numbers of inflammatory cells and mediators were detected to preliminarily explore the mechanism of SD in improving PE.Result:Compared with the conditions in the normal control group,the amount of pleural effusion,number of white blood cells in pleural effusion,lung wet/dry weight ratio,lung water content and lung permeability were increased(P<0.01)in the model group,where the rats presented cough,dyspnea,shortness of breath and arched back,and a small number of them had wet nose and bubble liquid in nostrils.In the autopsy of the rats in the model group,the lungs were enlarged or accompanied by congestion and plenty of pink bubble liquid appeared at the trachea.Compared with the conditions in the model group,SD reduced the amount of pleural effusion,number of white blood cells in pleural effusion,lung coefficient,lung wet/dry weight ratio and lung water content(P<0.01),and improved pulmonary edema symptoms such as damage,inflammation and infiltration around the lumen,thickening of the trachea,and accumulation of edema fluid,and the SD medium dose group had the best effect on the treatment of PE.In terms of respiratory system,compared with the normal control group,the model group had reduced latent time of cough and asthma(P<0.05,P<0.01)and elevated number of cough and wheezing(P<0.01).The three SD groups had increased latent time of cough and asthma and decreased number of cough and wheezing(P<0.01).In terms of urinary system,compared with the normal control group,the model group presented decreased urine volume.The SD low,medium and high dose groups had increased urine volume(P<0.05,P<0.01),but they had no effect on perspiration.In terms of digestive system,compared with the conditions in the normal control group,the gastric residual rate and gastrin(GT)level were increased(P<0.05,P<0.01),and the gastric emptying rate and small intestine transit rate were decreased(P<0.01).The SD low dose group had elevated small intestine transit rate(P<0.01),and the SD high and medium groups had enhanced gastric emptying rate and small intestine transit rate(P<0.01),reduced gastric residual rate,lowered GT level to promote gastrointestinal movement and transportation(P<0.01),and increased motilin(MTL)level to promote gastric emptying in rats(P<0.05,P<0.01).In terms of circulatory system,compared with the normal control group,the model group displayed reduced left ventricular ejection fraction(LVEF),left ventricular short axis shortening rate(LVFS)and cardiac output(CO)(P<0.01),and elevated tendency of heart rate,systolic blood pressure(SBP,P<0.01)and diastolic blood pressure(DBP,P<0.05).Compared with the model group,the SD low dose group increased LVEF and decreased DBP(P<0.05),while the SD medium dose group increased LVEF,LVFS,CO and SBP(P<0.01)and decreased DBP(P<0.05),and the SD high dose group increased LVFS(P<0.01)and decreased SBP(P<0.01)and DBP(P<0.05).In terms of central nervous system,compared with the conditions in the normal control group,the standing times dropped in the model group(P<0.01).SD reduced the movement distance,movement time,standing times and activity time in the center of the open field,and increased the rest time and activity time at the edge of the open field(P<0.05,P<0.01).Moreover,compared with the normal control group,the model group had serious inflammatory infiltration around the lung lumen,thickened trachea with accumulated edema fluid,seriously damaged lung tissue,increased number of white blood cells and percentage of neutrophils in blood(P<0.01),elevated percentage of monocytes,interleukin-4(IL-4),immunoglobulin E(IgE)level and reactive oxygen species(ROS)level in lung tissue(P<0.05,P<0.01),and decreased IFN-γ in alveolar lavage fluid(P<0.01).Compared with the model group,SD decreased the number of white blood cells,neutrophil accumulation,pulmonary congestion and interstitial edema,IFN-γ and IL-4 levels in alveolar lavage fluid and ROS level in lung tissue,and increased IgE level(P<0.05,P<0.01).Conclusion:SD had a significant improvement effect on PE model where the disease was located in the viscera.It could regulate the excretion of water by purgation,regulate Qi movement and expel Qi stagnation by descending and sinking lung Qi,and promote purification and descent of lung qi to make Qi movement downward.This indicated SD had the descending and sinking properties.The medium dose of SD decoction exerted the best effect,and its mechanism of action might be through regulating the neutrophil inflammatory response.