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补肾平喘方对肾虚哮喘模型大鼠的作用及其机制的实验初探

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.2028・ 安徽医药Anhui Medical and Pharmaceutical Journal 2013 Dee;17(12) 补肾平喘方对肾虚哮喘模型大鼠的作用及其机制的实验初探 高海玲 ,王迪生 ,龙子江 ,邢亚群 (1.蚌埠医学院,安徽蚌埠233000;2.蚌埠医学院第二附属医院药剂科,安徽蚌埠233004; 3.安徽中医药大学药理学教研室,安徽合肥230038) 摘要:目的观察补肾平喘方对肾虚致哮喘模型大鼠的实验影响并探讨其作用机制。方法SPF级清洁雄性大鼠60只,除空 白对照组外,其余大鼠均采用注射卵蛋白(OVA)与氢氧化铝致敏、OVA雾化吸入激发来复制哮喘动物模型,造模后将其随机分 为5组,模型组(等容量蒸馏水)、补肾防喘片组(377.5 mg・kg )、补肾平喘方高剂量组(49.68 g・kg )、补肾平喘方中剂量 组(24.84 g・kg )、补肾平喘方低剂量组(12.42 g・kg ),每组10只。并于致敏2周后开始灌胃给药8周。末次给药后,采 用ELISA法检测血浆ACTH,血清CORT的含量;取肺脏,一侧做HE染色,另一侧采用免疫组化法检测肺组织内t3-AR的含量。 结果补肾平喘方能有效缓解哮喘的症状,减轻肺组织病变程度,同时显著升高血清CORT,血浆ACTH,肺组织B—AR的水平。 结论补肾平喘方通过补肾温阳,可改善肾虚致哮喘模型大鼠的症状,其机制可能与改善HPA轴的功能有关。 关键词:肾虚;哮喘;肺组织;肾脏;补。肾平喘方 Effect and mechanism of the bushen pingchuan prescription for kidney.asthenia asthma in experimental rats GAo Hai.1ing ,WANG Di—sheng ,LONG Zi.jiang ,et al (1.Bengbu Medical College,Bengbu,Anhui 233000;2.Department ofPharmacy,The Second Afilfiated Hospital ofBengbu Medical College,Bengbu,Anhui 233004;3.Department ofPharmacology,Anhui University fChionese Medicine,Hefei,Anhui 230038) Abstract:Objective To observe the effect of the Bushen Pingchuan Prescription for kidney—asthenia asthma in experimental rats and to explore its mechanism.Methods Sixty clean male rats of SPF were chosen.In addition to the control group,the remaining rats were in— jected with ovalbumin(OVA)and aluminum hydroxide sensitization,OVA inhalation excited to replicate animal models of asthma.After the modeling rats were divided into five groups the model group(equal volume of distilled water),kidney anti-asthma tablets group (377.5 mg・kg ),Bushen Pingchuan Prescription high dose group(49.68 g・kg ),middle dose group(24.84 g・kg )and low dose group(12.42 g・kg ).Each group consisted of ten rats.The rats started to receive intragastric administration 2 weeks after sensi— tization,1 dose each day in 8 weeks.After the last administration plasma ACTH,serum CORT levels were detected by ELISA.One side of the lung was taken for HE staining,the other side was detected by immunohistochemistry in lung tissue 3-AR content.Results Bushen pingchuan prescription could effectively alleviate asthma,reduce asthma symptoms and improve lung tissue morphology,meanwhile,signif- icantly elevate seIum CORT,plasma ACTH,lung tissue B—AR levels.Conclusions Bushen pingchuan prescription improves asthma symptoms of asthma rat model caused by kidney deficiency which may be associated with improved HPA axis function. Key words:kidney;asthma;lung;kidney;bushen pingchuan prescription 支气管哮喘(简称哮喘)是由肥大细胞,嗜酸性粒细胞和 淋巴细胞参与的慢性持续性气道炎症性疾病,属于I型超敏 反应。临床上以痰、咳、喘为其主症,晚期多并发肺气肿、肺 心病。中医将该病归于“哮”证和“喘”证的范畴,对其病理机 制,中医认为本病以肺、脾、肾亏虚为本,以风、寒、湿、痰、瘀 为标 。 开发提供实验支持 1材料 1.1药物。 补肾平喘方由附子10 g(批号:120516)、甘草5 g(批号:121007)、淫羊藿15 g(批号:120627)、巴戟天10 g(批 号:120908)、决明子10 g(批号:120613)、白芍15 g(批号: 120305)、熟地20 g(批号:130128)、麻黄10 g(批号:120605)、 紫菀10 g(批号:121105)、青皮1O g(批号:130219)、桂枝10 g (批号:120810)、苏子10 g(批号:130303)、细辛3 g(批号: 120701)等中药组成,所有中药都按生药人与动物的剂量换 目前西医临床采取抗炎解痉为主的治疗原则,使用糖皮 质激素、B-受体激动剂及白三烯受体拮抗剂、茶碱等药物治疗 取得了一定的疗效,但远期疗效不很理想,同时也存在一定的 副作用。补肾平喘方是临床经验方,但对其作用机制还有待 探讨、研究。本实验通过观察补肾平喘方对实验大鼠血浆促 肾上腺皮质激素(ACTH),血清皮质酮(CORT),肺组织内B— AR的含量及肺脏病理学改变的研究,旨在探讨补肾平喘方 对哮喘大鼠的治疗作用及其相关机制,并为该处方的进一步 作者简介:高海玲,女,硕士研究生 通信作者:王迪生,男,副教授,硕士生导师,研究方向:中药新药开发 与研究,E-mail:wangdisheng2168@yahoo.com.ca 算,取生药按中药常规方法煎煮3次,合并药液,制成浓度为 5 g・ml 的水煎液。补肾防喘片,太极集团重庆涪陵制药厂 有限公司,批号1l120008。临用时配成相应浓度使用。 1.2动物4—5月龄SPF级未曾交配SD大鼠60只,雄性, 体重(200±20)g,由安徽医科大学动物中心提供,动物合格证 书号:SCXK(皖)2011—002。室温22~25℃、相对湿度为 40%一50%,普通饲料喂养,自由摄水。 1.3主要试剂卵清蛋白(Sigma公司进口分装);氢氧化铝 (江苏永华精细化学品有限公司,批号:20120909);甲醛(上 安徽医药Anhui Medical and Pharmaceutical Journal 2013 Dec;17(12) ・2031・ 双氢青蒿素抗乳腺癌MDA-MB一231细胞裸鼠移植瘤作用实验研究 李静 ,陈永顺 ,贾晓栋 ,梁建梅 ,杨斌 (1.商丘医学高等专科学校,河南商丘476100;2.广西医科大学药学院,广西南宁530021) 摘要:目的 研究双氢青蒿素(alpha—Dihydroartemisinin,DHA)对人乳腺癌MDA MB一23 1细胞裸鼠移植瘤的抗瘤作用。方法将 裸鼠随机分为阴性对照组、DHA低剂量组(DHA给药剂量为50 mg・kg~・d )、DHA中剂量组(DHA给药剂量为100 mg・ kg~・d。。)和DHA高剂量组(DHA给药剂量为200 mg・kg~・d-1),建立人乳腺癌MDA—MB一231细胞裸鼠移植瘤模型,测定 不同浓度的DHA对裸鼠的体重、抑瘤率、胸腺指数、脾脏指数的影响,免疫组化检测移植瘤P53蛋白的表达。结果DHA可明 显抑制移植瘤的生长,DHA低剂量、中剂量和高剂量对移植瘤的抑制率分别为20.25%、31.01%和46.20%,DHA各组胸腺指 数和脾脏指数较阴性对照组均有不同的的提高,与阴性对照组比较,DHA高、中剂量组小鼠胸腺指数和脾脏指数均有显著性差 异(P<0.05);免疫组化结果显示DHA可下调移植瘤细胞P53蛋白的表达。结论DHA对人乳腺癌MDA.MB-231细胞移植瘤 有显著的抑制作用,其抗癌机理可能是调节机体免疫和下调P53蛋白的表达。 关键词:双氢青蒿素;抗肿瘤 Antitumor effect and mechanism of dihydroartemisinin on breast cancer in nude mice with transplanted tumor of human MDA.MB.231 cells LI Jing ,CHEN Yong.shun ,JIA Xiao—dong .et al (1.Shangqiu Medical College,Shangqiu,Henan 476100,China;2.Guangxi Medical University,Nanning,Guangxi 530021,China) Abstract:Objective To observe the antitumor effect and mechanism of dihydroartemisinin on breast cancer in nude mice with trans- planted tumor of human MDA・MB一231 cells.Methods The nude mice were randomized into 4 groups:the negative control group,DHA low dose group,DHA middle dose group and DHA high dose group,then nude mice model was established with transplanted tumor of hu— man MDA—MB一23 1 cells.Different concentrations of DHA were used in nude mice with transplanted tumor of human MDA—MB-23 1 cells, whose effects on mice weight,inhibition rate of tumor,thymus index and spleen index were determined,and the expression of p53 protein was detected by immunohistochemical staining.Results Transplanted tumor inhibition rates in DHA low dose,middle dose and high dose groups were 20.25%,31.01%and 46.20%respectively.The thymus index and spleen index of DHA group were improved.the thymus index and spleen index in the high and middle dose groups were signiifcantly different compared with negative control group. Conclusion DHA could signiifcantly inhibit the growth of transplanted tumor of human MDA—MB一23 1 cells,whose mechanism may be through adjusting the body’s immunological function and down—regulating the expression of p53 protein. Key words:DHA;anti—tumor activity 乳腺癌是一种严重危害妇女健康的恶性肿瘤,在西方发 Bromide,MTr)购自Sigma公司,DHA(四川三奇制药,纯度为 达国家已成为妇女的第一位死亡原因,在我国也是女性最常 99%),p53一抗(南京建成生物工程研究所)。 见的恶性肿瘤,且发病率呈逐年上升的趋势 。乳腺癌是一 1.2主要仪器全自动CO:孵箱(Themro Formo),酶联免疫 种对化疗药物敏感的肿瘤,但常用的乳腺癌化疗药物毒性大, 检测仪(Bio—Rad),病理图像分析仪(德国Leiea),电子天平 患者常常无法忍受为化疗所承受的巨大痛苦,因此,筛选低毒 JA2003(上海精密仪器厂)。 高效的抗癌药物显得尤为重要。 1.3实验动物SPF级小鼠40只,5~6周龄,雌性,体重(20 1材料和方法 ±2)g,由商丘医学高等专科学校实验动物中心提供(许可证 1.1主要试剂RPMI.1640培养液(美国Gibco公司),小牛 号SQYZ-2Ol3-B02)。 血清(杭州四季青公司),四唑盐(Thiazolyl Blue Tetrazolium 1.4细胞株和细胞培养人乳腺癌MDA—MB-231细胞株购 参考文献: [6] 王培,许利平,金喜宏,等.糖皮质激素对哮喘模型大鼠肾阴阳 [1]任现志,刘嘉君.从肾论治儿童哮喘缓解期的研究概况[J].中 虚证候演变的影响[J].中西医结合学报,2010,8(1):8O一85. 国中医急症,2010,19(12):2120—2121. 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