【作者】 王贤喜；

【导师】 明亮； 杨解人；

【作者基本信息】 安徽医科大学 ， 药理学， 2004， 硕士

【摘要】 目的： 探讨晕可定颗粒的主要药效学，为临床提供药效学资料及治疗学基础。 方法： 1．豚鼠随机分组：①模型对照组；②给药组(低剂量组、中剂量组和高剂量组)，分别ig给药晕可定4、8、16g／kg：③阳性对照组，给予抑眩宁口服液。三组动物均采用三氯甲烷破坏豚鼠一侧膜迷路感受器模型，从注射入三氯甲烷后20min、40min和60min观察、记录眼震颤和头左右摇摆次数，每次记录1min为准，旋转只记录1h内总次数。探讨受试药对眼震颤、摆头及旋转的影响； 2．实验动物随机分为：①积水模型组，行内淋巴囊和内淋巴管阻塞手术，制造豚鼠膜迷路实验性内淋巴积水模型；②给药模型组，在造模的基础上给予ig给药晕可定8g／kg：③假手术组。取内耳组织，采用石蜡和火棉胶两种包埋和切片方法，观察造模后及用药后对内耳组织SMA增加率等形态学的影响； 3．选用家鸽随机分组：①空白对照组；②给药组(低剂量组、中剂量组和高剂量组)分别ig给药晕可定4、8、16g／kg，连续3d给药；③阳性药组(抑眩宁0.63g／kg)。各组均采用硫酸铜和顺铂致家鸽呕吐的模型，观察受试药的镇吐作用； 4．小鼠随机分组：①空白对照组；②YKD给药组(低剂量组、中剂量组和高剂量组)分别ig给药晕可定5、10、20g／kg；③阳性药组(抑眩宁0.9g／kg)。采用阈剂量戊巴比妥钠(50mg／kg)致小鼠睡眠法，研究受试药的催眠作用；以翻正反射消失为入睡时间，从翻正反射消失至恢复时间为睡眠持续时间。 5．小鼠分组同上，YKD给药组和阳性药组连续给药7d，末次给药后1小时，尾静脉注射0.5％伊文思蓝生理盐水溶液，然后冰醋酸(Hac)—生理盐水腹腔注射诱导毛细血管渗出，再用生理盐水分数次洗涤腹腔，最后洗涤液上清在590nm波长进行比色测定，研究受试药对小鼠腹腔毛细血管通透性的影响； 6．SD大鼠随机分组：①空白对照组；②YKD给药组(低剂量组、中剂量组和高剂量组)分别ig给药晕可定4、8、16g／kg；③阳性药组。采用大鼠代谢笼法，每只安徽医科大学硕士学位论文代谢笼放一只大鼠，收集并记录给药后12h尿量，观察受试药的利尿作用;7.小鼠随机分组基本同上:①空白对照组;②YKD给药组(低剂量组、中剂量组和高剂量组)分别19给药晕可定5、10、209/kg;③阳性药组。末次给药后1小时，把小鼠投入箱中适应5分钟后，记录2分钟内小鼠走动时间及双前肢向上抬举次数，观察小鼠自发活动的改变，研究受试药的镇静作用;8.采用中药血清药理学方法，c57BL/6)小鼠按5、10、209众g体重灌胃晕可定颗粒，于给药后Zh取血分离含药血清。蒸馏水灌胃取小鼠空白血清。另取C57BL/6)小鼠股骨，细胞因子分离骨髓前体细胞，分设3组进行培养:①空白对照组，培液内加入GM一CSF和IL一4常规培养。②在空白对照的基础上，加入空白血清。③在空白对照的基础上，加入晕可定颗粒含药血清。用倒置相差显微镜及Gimesa’染色、免疫组织化学观察三组体外培养细胞向树突状细胞分化情况及MHCH类分子(I一Ab)的表达变化，探讨晕可定颗粒含药血清对免疫抗原呈递细胞的影响。结果:1.三氯甲烷微量注射可破坏右侧膜迷路感受器，模型空白对照组豚鼠出现步态不稳，眼球震颤，身体向患侧旋转，头向左右摇摆;抑眩宁灌胃阳性对照组、晕可定颗粒(8、169/kg)灌胃模型给药组豚鼠眼球震颤次数减少，与对照组比较，差异有显著意义(尸<0.05);但晕可定模型给药组豚鼠头摆动和身体旋转变化无显著差别;2.通过内淋巴囊和内淋巴管阻塞手术，成功复制不同程度内淋巴积水豚鼠模型，与假手术组比较，表现为前庭膜重度膨出，前庭阶缩小，膜蜗管增大，SMA增加率变大，差异有显著意义(尸<0.01);晕可定颗粒灌胃后可减轻豚鼠实验性内淋巴积水的程度，前庭膜轻度膨出，与造模组比较，SMA增加率差异有显著意义(P<0.01);但与空白对照组比较SMA增加率差异无显著意义(尸>.05)。3.硫酸铜溶液给家鸽灌胃及翼下静脉注射顺铂，都可导致家鸽呕吐，晕可定颗粒 (4、8、169/kg)可明显减少硫酸铜致家鸽呕吐的次数，晕可定颗粒(4、89/kg)灌胃可明显延长硫酸铜致吐的潜伏期(P<0.05或尸<0.01);4.晕可定颗粒(5、10、209/kg)可明显缩短戊巴比妥钠致小鼠睡眠的入睡时间，安徽医科大学硕士学位论文按10、209/kg晕可定颗粒灌胃还可延长小鼠睡眠持续时间(尸<0.05或尸<0.01);5.晕可定颗粒(10、209/kg)可以抑制腹腔注射冰醋酸所致的毛细血管通透性增加，与空白对照组比较，差异显著(尸<0.01);6.用代谢笼法记录12小时大鼠尿量，观察到晕可定颗粒无明显利尿作用;7.按5、10、20眺g体重给小鼠灌胃不同剂量晕可定颗粒，均可显著减少小鼠走路时间和举前肢次数，对小鼠自发活动有抑制作用;8.加入血清后可促进骨髓前体细胞向树突状细胞分化、成熟，表现为DC集落增多增大，细胞有许多明显的突起，I一A”表达增加;但晕可定颗粒含药血清组与空白血清组比较差别不显著。结论: YKD颗粒可以降低毛细血管的通透性，减轻内淋巴囊积水程度，还具有镇静催眠镇吐作用，因此对梅尼埃病有对抗治疗作用。但对免疫性梅尼埃病有无对因治疗作用，仍需进一更多还原

【Abstract】 Objective:To observe and assess the pharmacological effect of Yunkeding Granule(YKD) on Meniere’s disease (MD). Methods:1 .Using the damage model of membranous labyrinth sensor induced by chloroform to observe the effect of YKD Granule on nystagmus as well as head swing and circumrotating.2.Using the model of Endolymphatic hydrops induced by surgery, to study the therapeutical function of YKD Granule on the morphology of inner ear. SMA increasing rate was also examined.3.The effect of YKD Granules on the pigeon vomiting induced by bluestone as well as by cisplation was also studied.4.The effect of YKD Granules on the mouse dormancy induced by low dose sodium pentobarbital was observed too.5.To study the influence of YKD Granules on the abdominal capillary permeability, Evan’s blue injection was adopted.6.Using rat metabolizing baskets, the effect of YKD Granules on the urine output was observed.7.The effect of YKD Granules on mouse spontaneous activities was analysised. 8.To look into the effect of YKD Granules on the cloning of BMDC in vitro and its MHC Class II molecule expressin, we adopted a new method of the serum pharmacology. BMDCs were isolated from C57BL/6J mice and cultured in RPMI1640 complete medium with GM-CSF plus IL-4. BMDCs were grouped:(1)normal control;(2) control serum, additionally adding blank serum; (3) Drugs containing serum,additionally adding blank serum. The different morphology characteristics of BMDC indifferent stages were observed with microscope and Gimesa’ dyeing as well asimmunocytochemistry analysis.Results:1 .Chloroform damaged the right membranous labyrinth sensor.The Guinea-pigs appeared nystagmus and circumgyration as well as head swing.Compared to control group animals ,YKD Granule (8, 16g/kg) could obviouslyreduce nystagmus degree (P<0.05), but had no effect on circumrgyration and headswing.2.The model of, Endolymphatic hydrops was successfully made by surgery.Compared to control, Guinea-pigs of surgery model group were found that theirvestibular membranes were obviously protruding and stria vascularis were abnormitytoo.However the vestibular membranes of animals i.g.with YKD Granule were slightlyprotruding and the morphological characteristics of stria vascularis was also resumed.Moreover it was founded that the difference of SMA increasing rate between groupswas remarkable (P<0.01).3.Both bluestone and cisplation could induce pigeon vomiting. YKD Granules couldobviously reduce vomiting degree and prolong the delitescence4.YKD Granules ( 5 10 20g/kg)could shorten the time span before dormancy inducedby sodium pentobarbital and prolonged dormancy lasting period at the dose of 1020g/kg i.g. (compared to control: P<0.05 or P<0.01).5.After Evan’s blue injection , the absorbency of abdominal washing fluid wasexamined at 590nm. Compared to control we found that YKD Granules (10 20g/kg )could restrain the increase of abdominal capillary permeability (P<0.0l).6.YKD Granules was observed to have no effect on the urine output by using ratmetabolizing basket, The difference in groups was not statistically significant. . (compared tocontrol: P>0.05)7.YKD Granules (5 , 10, 20g/kg ) could significantly cut down mouse spontaneousactivities such as walking and lifting forelimb.8.Both control serum and drugs containing serum could accelerate the differentiationand maturing of BMDC, inducing BMDC with many more long processes and I-Aexpression by immunocytochemistry analysis. But there was no obvious differencebetween control serum and drugs containing serum.Conclusion:YKD Granules had obvious effect of calmness and suppressive function on vomiting.ltcould counterwork some symptoms of model animals with Endolymphatic hydrops byreducing abdominal capillary permeability and lighting the degree of endolymphatichydrops and so on. But it perhaps had no function to the immunological mechanisms in thepathogenesis of Menie更多还原