天南星和制天南星对过敏性哮喘大鼠的影响比较  

Comparison on Ameliorating Effect of Arisaematis Rhizoma and Arisaematis Rhizoma Preparatum on Allergic Asthma in Rats

作　　者：邹婷 王晶[1] 武旭 杨凯 贺素容 钟恋 王昌利[1] 赵重博[1,3,4] ZOU Ting;WANG Jing;WU Xu;YANG Kai;HE Surong;ZHONG Lian;WANG Changli;ZHAO Chongbo(Shaanxi University of Chinese Medicine,Xianyang 712046,China;Sichuan Institute for Drug Control,Chengdu 610032,China;Engineering Technology Research Center of Shaanxi Administration of Chinese Herbal Pieces,Xianyang 712046,China;Shaanxi Traditional Chinese Medicine Processing Technology Heritage Base,Xianyang 712046,China)

机构地区：[1]陕西中医药大学,陕西咸阳712046 [2]四川省药品检验研究院,成都610032 [3]陕西省中药饮片工程技术研究中心,陕西咸阳712046 [4]陕西省中药炮制技术传承基地,陕西咸阳712046

出　　处：《中国实验方剂学杂志》2024年第1期120-129,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金青年基金项目(81803732);陕西省中药炮制技术传承基地建设项目(138010015);中国药品监管科学行动计划项目(NMPAJGKX-2023-077);四川省药品检验研究院院立科研项目(2023-KYYL-001);四川省自然科学基金项目(2022NSFSC1572)。

摘　　要：目的:考察生姜-白矾炮制天南星前后对过敏性哮喘大鼠的疗效及作用差异,为制天南星“炮制增效”理论提供依据。方法:70只SD大鼠采用卵清蛋白(OVA)-氢氧化铝腹腔注射建立过敏性哮喘大鼠模型,通过天南星(1.2、0.3 g·kg^(-1)·d^(-1))和制天南星(1.2、0.3 g·kg^(-1)·d^(-1))水提液灌胃,阳性药使用孟鲁司特钠(0.001 g·kg^(-1)·d^(-1)),通过考察大鼠血清及肺泡灌洗液(BALF)中辅助性T细胞1型细胞群/2型细胞群(Th1/Th2类细胞因子)与BALF中炎细胞分类计数比例变化;实时荧光定量聚合酶链式反应(Real-time PCR)检测肺组织中黏蛋白5AC(MUC5AC)的mRAN表达情况;苏木素-伊红(HE)染色、过碘酸-雪夫(PAS)染色观察大鼠肺组织病理变化;免疫组化法观察大鼠肺组织中c-Jun氨基末端激酶(JNK)、细胞外调节蛋白激酶(ERK)和p38丝裂原活化蛋白激酶(p38 MAPK)蛋白表达,通过蛋白免疫印迹法(Western blot)检测肺组织ERK、p-ERK、JNK、p-JNK、p38MAPK、p-p38 MAPK蛋白表达,采用总评归一分析比较天南星和制天南星的药效差异。结果:与空白组比较,模型组大鼠血清和BALF中白细胞介素-12(IL-12)、γ干扰素(IFN-γ)的含量明显降低(P<0.05,P<0.01),白细胞介素-4(IL-4)、白细胞介素-5(IL-5)、白细胞介素-13(IL-13)含量明显升高(P<0.05,P<0.01);与模型组比较,孟鲁司特钠组、天南星高剂量组和制天南星高剂量组大鼠血清和BALF中IL-12、IFN-γ含量明显升高(P<0.05,P<0.01),IL-4、IL-5、IL-13含量明显降低(P<0.05,P<0.01),天南星低剂量组和制天南星低剂量组大鼠血清中IFN-γ含量明显升高(P<0.05),IL-4、IL-13明显降低(P<0.05,P<0.01),BALF中IL-5、IL-13含量明显降低(P<0.05,P<0.01);BALF中巨噬细胞、淋巴细胞、中性粒细胞、嗜酸性粒细胞百分比降低;并抑制肺组织中JNK/ERK/p38 MAPK通路和MUC5AC蛋白的表达。总评归一分析发现,同剂量给药后,制天南星组药效略优于天南星组。结论:天南星和制Objective:To investigate the therapeutic effects and difference in the effects of Arisaematis Rhizoma(AR)before and after processing(i.e.,Arisaematis Rhizoma Preparatum,ARP)with Zingiberis Rhizoma Recens-Alumen on allergic asthma in rats and to provide a basis for the theory of processing improving the efficacy.Method:A rat model of allergic asthma was established in 70 SD rats by intraperitoneal injection of ovalbumin(OVA)-aluminum hydroxide.The rats were administrated with the aqueous extracts of AR(1.2,0.3 g∙kg^(-1))and ARP(1.2,0.3 g∙kg^(-1))aqueous extracts by gavage,and montelukast sodium(0.001 g∙kg^(-1))was used as the positive drug.The T helper cell type 1/type 2(Th1/Th2)ratio in the serum and bronchoalveolar lavage fluid(BALF)and percentages of inflammatory cells in BALF were determined.Polymerase chain reaction(PCR)was employed to determine the mRNA level of mucin 5AC(MUC5AC)in the lung tissue.The pathological changes in the lung tissue were observed by hematoxylin-eosin(HE)staining and PAS staining.Immunohistochemical assay was employed to measure the expression of c-Jun amino-terminal kinase(JNK),extracellular signal regulated protein kinase(ERK),and p38 mitogen-activated protein kinase(p38 MAPK)in rat lung tissue.Western blot was employed to determine the protein levels of ERK,p-ERK,JNK,p-JNK,p38,p-p38 in the lung tissue.The effects of AR and ARP were compared based on overall desirability.Result:Compared with the blank group,the levels of interleukin-12(IL-12)andγinterferon(IFN-γ)in serum and BALF of rats in the model group were significantly lower(P<0.05,P<0.01),and the levels of interleukin-4(IL-4),interleukin-5(IL-5)and interleukin-13(IL-13)were significantly higher(P<0.05,P<0.01).Compared with the model group,the serum and BALF contents of IL-12 and IFN-γin rats in the montelukast sodium group,high-dose AR group and high-dose ARP group were significantly higher(P<0.05,P<0.01),and the contents of IL-4,IL-5 and IL-13 were significantly lower(P<0.05,P<0.01),and the serum contents of IFN-γi

关 键 词：天南星 制天南星 过敏性哮喘 辅助性T细胞(Th)1/Th2类细胞因子 丝裂原活化蛋白激酶(MAPK)通路 

分 类 号：R2-0[医药卫生—中医学] R33R289R256.12R781.67