节点文献

咽舒饮泡腾片治疗咽炎的药效学和帮得孕改善着床的实验研究

Effect of Yanshuyin Effervescen Tablets on Pharyngitis and Bangdeyun on EID

【作者】 徐婧

【导师】 吴云霞;

【作者基本信息】 华中科技大学 , 中药学, 2010, 硕士

【摘要】 目的:本研究旨在通过药理实验评估咽舒饮泡腾片治疗急慢性咽炎的药理药效和急性毒性情况,为进一步开发和扩大运用奠定基础。方法:采用二甲苯致小鼠耳廓肿胀,小鼠琼脂肉芽肿实验考察咽舒饮泡腾片的抗炎作用;采用鸡红细胞致小鼠迟发型超敏反应实验,考察咽舒饮泡腾片的抗过敏作用;采用小鼠气管酚红排泌的实验,考察咽舒饮泡腾片的滋阴化痰作用;观察咽舒饮泡腾片对乙型溶血性链球菌,肺炎双球菌,金黄色葡萄球菌,大肠杆菌,绿脓杆菌5种较常见易感菌的体外抑菌和杀菌情况。并将咽舒饮泡腾片以最大浓度最大剂量灌胃,连续7 d观察小鼠急性毒性反应情况。结果:表明咽舒饮泡腾片可明显抑制二甲苯所致小鼠耳廓肿胀,抑制琼脂肉芽肿组织增生和鸡红细胞所致迟发型超敏反应,在实验剂量范围内呈剂量依赖性;在一定剂量时能抑制肉芽肿组织增生和使酚红排出量增多,具有广谱抑菌作用并对乙型溶血性链球菌显示出非常良好的抑菌作用和杀菌活性。在咽舒饮泡腾片最大浓度、最大剂量灌胃后动物表现有活动减少,摄食减少,稀便甚至腹泻,但停药23d后均恢复正常,7 d后全部动物健存,体重情况与正常组无明显区别,未见其它任何毒副反应。结论:咽舒饮泡腾片的毒性较低,具有较好的抗炎、抗过敏和一定的抑菌杀菌与滋阴化痰药理作用,对治疗咽炎有良好的效果。目的:建立一种新的小鼠胚胎着床障碍模型并探讨其机制,以用来筛选促进着床药物,该模型既能高度有效的模拟临床,而且方法简便。并初步观察帮得孕对该着床障碍模型小鼠着床的改善。方法:仿照临床超排卵方案,运用腹腔注射孕马血清促性腺激素(pregnant mare serum gonadotropin,PMSG),并于48小时后注射绒毛膜促性腺激素(human chorionic gonadotrophin,HCG)的方法。分别观察5IU,7.5IU,10IU不同剂量PMSG和HCG方案下胚泡着床率和平均胚泡着床点数情况,确定超排卵对着床的影响并确定最佳造模方案和剂量。确定剂量后,观察该超排卵方案致胚泡着床障碍的可能机制。通过HE染色观察第4,5,6天子宫内膜组织形态学变化;放射免疫学方法检测血清中雌、孕激素含量;免疫组化方法观察子宫内膜雌、孕激素受体(雌激素受体estrogen receptor,ER;孕激素受体progestogerone receptor,PR),NF-κB及CD31的表达;伊文思蓝染色观察子宫内膜血管通透性情况。运用COH致小鼠胚泡着床障碍模型,受精当日低、中、高剂量分按6 g·kg-1,12 g·kg-1,24 g·kg-1(相当于生药材)剂量开始灌胃,直至妊娠D8,观察不同剂量的帮得孕的对着床的改善作用。结果:5IU,7.5IU,10IU不同剂量PMSG和HCG方案中,10 IU PMSG和HCG不同日注射方案既降低着床率,且明显降低平均着床点数,因此作为小鼠胚胎着床障碍模型的超排卵药物方案。该超排卵方案致胚泡着床障碍的可能机制为:1)超排卵使血清中,雌、孕激素水平明显上调且引起激素紊乱。正常小鼠雌、孕激素水平均于D4达高峰,超排卵后雌、孕激素峰值均出现在了D5,两组雌、孕激素的比值,正常组的比值在D6达到高峰,而超排卵组在D7天达到峰值,超排卵使ER时空转换表达延迟、减弱或缺失,但PR表达未见明显影响;2)超排卵引起子宫内膜蜕膜化较迟缓;3)超排卵可能干扰了母体对胚泡的免疫耐受形成。主要表现在NF-κB在正常组D6呈弱表达,超排卵组D6仍呈较强表达,而NF-κB是引起炎症相关因子表达转录的核转录因子;4)超排卵可能影响了早期胎盘血管生成。主要表现在超排卵使D4D6伊文思蓝染色减弱,表明对血管通透性有影响,且血管标志物CD31在正常组D6在胚泡周围开始增强表达,超排卵组D6在胚泡周围表达较弱。帮得孕6 g·kg(-1024 g·kg-1剂量范围内均显示出明显的促进着床的效果。结论:10 IU PMSG和HCG不同日注射方案作为小鼠胚胎着床障碍模型的超排卵药物方案,该模型符合临床实际,操作程序简便,结果重复性强。COH致小鼠胚泡着床障碍的机制可能与引起雌、孕激素比例失调,引起子宫内膜蜕膜化迟缓,干扰了母体对胚泡的免疫耐受形成和影响早期胎盘血管生成相关。帮得孕能够促进COH致胚泡着床障碍的小鼠着床,有效提高妊娠率。

【Abstract】 Objectives: To assess the main effect on pharyngitis and the acute toxicity of Yanshuyin Effervescent Tablets for further usage.Methods: Model of dimethylbenzene induced ear edema and agar-pill-implantation- induced hyperplasia in mice were used to observe the anti-inflammatory effects of the tablets; Chicken red blood cells induced mice delayed-type hypersensitivity experiment was used to observe its effect of anti-allergic; Mice trachea phenol red excretion experimental method was used to observe the role of expelling phlegm; minimal inhibitory concentration and minimal bactericidal concentration of Yanshuyin to beta-hemolytic streptococcus, diplococcus pneumoniae , staphylococcus aureus, E.coli, pseudomonas aeruginosa, five commonly susceptible bacteria in vitro were also observed. The mice with the largest concentration and the largest volume of oral administration of Yanshuyin reduced their activities, ate less and gave thin faeces or even diarrhea, but two or three days later after stoping ig they returned to the normal. Seven days later, no death of the animals was found, the average body weight of mice had no significant difference compared with the normal group, and there were no other toxic reactions.Conclusion: Yanshuyin have little toxicity, it has a good anti-inflammatory, anti-allergic, anti-bacteria effect and some expelling phlegm pharmacological effects. It is suitable for the treatment of pharyngitis. Objectives: Establish a new kind of EID model induced by COH, which can really close to the clinical, easy to operate and for drug screening, and explored its mechanisms. Then observed the effect of improving implantation of Bangdeyu on EID mice.Methods: Following the clinical method, treated with HCG 48h after PMSG treatment. Using 5IU, 7.5IU, 10IU dose of PMSG and HCG, observe the implantation rate and number of average implanted blastocyst spot to decide a best method and dose for the model. The doses decided, explored the probable mechanisms of the EID induced by COH. Using HE stain view the endometrium morphological change on day 4 to 6 of pregnancy; radioimmunoassay method test the contents of estrogens and progestin in mice serum; immunohistochemical method observe the expression of ER, PR, NF-κB, CD31 of endometrium; use evans blue dying view the vasopermeability of the endometrium. Using the EID model induced by COH, on the day of impregnation ig low, medium, high does 6 g·kg-1,12 g·kg-1,24 g·kg-1, till the day 8 of pregnancy, to observe the improving effect of different does Bangdeyun on implantation.Results: In different doses of 5IU,7.5IU,10IU PMSG and HCG, the 10IU doses which injected in different days could both reduce the implantation rate and average implanted blastocyst spot, thus chose it for the EID model dose induced by COH. The mechanisms of EID by this COH maybe: 1) COH affects the concentration of estrogens and progestin in mice serum and their receptors. Compared with normal mice serum concentration of estrogens and progestin peaked at day 4 of pregnancy, COH group peaked at day 5 of pregnancy, ratio of estrogens/progestin, normal mice peaked at day 6 of pregnancy, COH mice peaked at day 7, ER expression was delayed in time and space, weaked or lack, but PR had no obvious change; 2) COH delayed endometrium decidualization; 3) COH may interfere the mother’s immune tolerance to blastocyst, mainly manifested NF-κB expressed higher at day 6 of pregnancy than in normal group, NF-κB is a nuclear transcription fator which induce inflammatory factor express and transcription; 4) COH may affect the early placenta angiogenesis. It manifested mainly the evans blue dying of mice endometrium lighter on day 4 to 6 of pregnancy in COH group than in normal group, which means COH may affect the vascular permeability; and in endometrium of normal mice vessels, CD31 began highly expressed around the blastocyst, but COH mice expressed lower. Bangdeyun in 6 g·kg-1 to 24 g·kg-1 doses all showed obvious promoting effect on embryo implantation.Conclusion: 10IU does PMSG and HCG inject in different day can be used for inducing EID model in COH method, which is close to the clinical, easy to operate and can be replicated well. The mechanisms of EID model induced by COH may associated with leading to estrogens/progestin ratio out of control, endometrium decidualization delayed, interfering the mother’s immune tolerance to blastocyst and affecting the early placenta angiogenesis. Bangdeyun can enhance the mice implantation rate of EID induced by COH.

节点文献中: 

本文链接的文献网络图示:

本文的引文网络