发酵炮制对胆南星药性与刺激性的影响

Effects of drug properties and irritation in fermentation and processing of Arisaema Cum Bile

目的:通过比较天南星发酵炮制成胆南星的药性与刺激性变化,为胆南星发酵炮制“减毒改性”的传统理论提供现代药理学研究依据。方法:建立慢性阻塞性肺疾病(COPD)寒饮蕴肺证大鼠模型和慢性阻塞性肺疾病加重期(AECOPD)热痰证大鼠模型,给药组按照2.52 g/kg剂量给药,持续给药7 d,观察大鼠体温、体质量变化,ELISA法测定大鼠血清中中性粒细胞弹性蛋白酶(NE)、肾上腺素(E)、皮质醇(Cor)、C-反应蛋白(CRP)和支气管肺泡灌洗液中肺组织黏蛋白5AC(MUC5AC)、白细胞介素(IL)-8、肿瘤坏死因子-α(TNF-α)、嗜酸性粒细胞(EOS)、中性粒细胞(NEU)的含量,HE染色观察肺组织病理学状态。采用CCK-8检测不同药物浓度对HaCaT细胞增殖的影响,ELISA法检测HaCaT细胞上清液中IL-1β、IL-8、转化生长因子-β1(TGF-β1)、细胞间黏附分子-1(ICAM-1)、前列腺素E2(PGE2)含量,考察药物对细胞刺激性的影响。结果:在COPD寒饮蕴肺证大鼠模型中,与模型组比较,天南星组能显著降低NE、IL-8、TNF-α、MUC5AC、EOS含量(P<0.01),胆南星组治疗效果不明显。在AECOPD热痰证大鼠模型中,与模型组比较,胆南星组能显著降低E、NE、Cor、CRP,IL-8、TNF-α、MUC5AC含量(P<0.01),增加NEU含量(P<0.01),天南星组治疗效果不明显。与空白对照组比较,天南星组各浓度能显著升高HaCaT细胞中IL-1β、IL-8、TGF-β1、ICAM-1、PGE2含量(P<0.01),而胆南星组较天南星组各浓度上述指标均显著降低(P<0.01)。结论:天南星经胆汁发酵炮制成胆南星后,功效由温化寒痰转化为了清热化痰且毒性降低,验证了天南星发酵炮制成胆南星的“减毒改性”传统理论。

Objective:By comparing the drug properties and irritating changes of Arisaematis Rhizoma and Arisaema Cum Bile,to provide modern pharmacological research basis for the traditional theory of‘reducing toxicity and changing drug properties’of Arisaema Cum Bile fermentation and processing.Methods:A rat model of chronic obstructive pulmonary disease with the cold-phlegm in the lung syndrome(COPD-CPS)and acute exacerbation of chronic obstructive pulmonary disease with the heat-phlegm syndrome(AECOPD-HPS)was established,respetively,the drug groups were administered at a dose of 2.52 g/kg,continuously administered for 7 days,the body temperature and body mass changes of rats were observed.Epinephrine(E),the neutrophil elastin(NE),cortisol(Cor)and C-reactive protein(CRP)levels in rat serum and lung tissue mucin(MUC5AC),interleukin(IL)-8,tumor necrosis factor-α(TNF-α),eosinophils(EOS),neutrophils(NEU)in bronchoalveolar lavage were determinated by ELISA.Pathological state of lung tissue was determinated by HE staining.The effects of different drug concentrations on cell proliferation were detected by CCK-8,and the effects on cell irritation were investigated by ELISA method using the HaCaT cell supernatant inflammatory factors IL-1β,IL-8,transforming growth factor-β1(TGF-β1),serum soluble intercellular adhesion molecule-1(ICAM-1),and prostaglandin E2(PGE2).Results:In the model of COPD-CPS,compared with the model group,the Arisaematis Rhizoma group could significantly reduce NE,IL-8,TNF-α,MUC5AC and EOS(P<0.01),the therapeutic effect of Arisaema Cum Bile group was not significant.In the model of AECOPD-HPS,compared with the model group,the Arisaema Cum Bile group significantly reduced the content of E,NE,Cor,CRP,IL-8,TNF-α,MUC5AC(P<0.01),and increased the content of NEU(P<0.01),the therapeutic effect of Arisaematis Rhizoma group was not significant.Compared with the blank control group,different concentration of the Arisaematis Rhizoma group significantly increased the contents of IL-1β,IL-8,TGF-β1,ICAM-1,and PGE2 in HaCaT cells(P<0.01),while the above indicators were significantly reduced in all concentrations in the Arisaema Cum Bile group compared to the Arisaematis Rhizoma group(P<0.01).Conclusion:After Arisaematis Rhizoma is processed into Arisaema Cum Bile by bile fermentation,its efficacy is transformed from warming and resolving cold phlegm to clearing away heat and resolving phlegm,and its toxicity is reduced,which verifies the traditional theory of‘reducing toxicity and changing drug properties’of Arisaematis Rhizoma fermentation and processing into Arisaema Cum Bile.