【作者】 张淑二；

【导师】 章孝荣； 陶勇； 方富贵；

【作者基本信息】 安徽农业大学 ， 动物遗传育种与繁殖， 2008， 硕士

【摘要】 以山羊胎盘为原料,用现代工艺进行提取,并将提取物(GPE)与其他辅助原料按照不同的配方分别制备成羊胎盘复方制剂Ⅰ号(GPPⅠ)和羊胎盘复方制剂Ⅱ号(GPPⅡ),在两种复方制剂中,每500mg制成品的羊胎盘含量相当于鲜胎盘30g。以小鼠为实验动物,对山羊胎盘提取物及两种不同配方的复方制剂进行毒理学检测(包括急性毒性实验、30天喂养实验、鼠伤寒沙门氏菌/哺乳动物微粒体酶实验、睾丸染色体畸变实验、骨髓细胞微核实验)、免疫功能测定(包括ConA诱导小鼠脾淋巴细胞转化实验和小鼠NK细胞活性检测实验)和缺氧耐受力实验(包括常压缺氧实验、亚硝酸钠中毒存活实验和急性脑缺血性缺氧实验)。实验中,GPE、GPPⅠ和GPPⅡ在每个实验中各设一个单倍剂量组,以便在先前研究的基础上再进一步确认GPPⅡ的配方优势,并且再在每个实验中根据相关检测要求,对GPPⅡ设计低、中、高三个剂量组和对照组,每组小鼠数量为5～20只(按实验的具体要求而定),各实验组小鼠均采取人工灌胃方法投喂产品。研究结果如下:1、在急性毒性实验中,各剂量组小鼠采食及排便正常,精神状态良好,均未出现异常表现及死亡现象,。2、在30天喂养实验中,各剂量组小鼠精神状况良好,生理行为正常,未出现中毒迹象和死亡;在食物利用率、各项血液学和血液生化指标均无显著变化(P＞0.05),各器官组织病理检查未发现病变和细胞组织异样,绝大部分重要脏器的绝对重量与相对重量比差异不显著(P＞0.05),但GPPⅡ部分剂量组脾脏的绝对重量与相对重量比与对照组相比有显著差异(P＜0.05)。3、在鼠伤寒沙门氏菌/哺乳动物微粒体酶实验(Ames实验)实验中,GPE、GPPⅠ和GPPⅡ各剂量组的回变菌落数,不论加S-9与否,均未超过相应溶剂对照组值的两倍,同时也未发现剂量—反应关系。故羊胎盘各制剂的Ames实验结果为阴性,表明无致突变作用。4、在小鼠睾丸染色体畸变实验中,GPE、GPPⅠ及GPPⅡ的各剂量组小鼠睾丸染色体的断片数、易位数、染色体(包括常染色体和性染色体)单价数、畸变细胞数及其畸变率与阴性对照组相比差异不显著(P＞0.05),但与CTX处理的阳性对照组达到极显著差异(P＜0.01)。5、在小鼠骨髓细胞微核实验中,GPE、GPPⅠ及GPPⅡ的各剂量组小鼠骨髓PCE微核率与阴性对照组相比无显著性差异(P＞0.05)。而阳性对照组的小鼠骨髓PCE微核率则显著高于其他实验组(P＜0.01)。6、免疫功能检测中,小鼠脾淋巴细胞转化实验和NK细胞活性实验结果表明,GPPⅠ实验组与GPPⅡ单、中剂量组的小鼠的脾淋巴细胞增值能力显著增强(P＜0.05),GPPⅡ低、高剂量组小鼠脾淋巴细胞增值能力达到极显著水平(P＜0.01),GPE则差异未达到显著水平(P＞0.05)。GPPⅡ低、高剂量组小鼠脾淋巴细胞增殖能力也显著高于GPEⅠ号和GPE组(P＜0.05);GPPⅡ单、低、中剂量组小鼠的NK细胞活性显著增强(P＜0.05),且低剂量组差异极显著(P＜0.01),其他实验组差异不显著(P＞0.05)。7、缺氧耐受力实验结果表明,与对照组相比,GPPⅡ单、中、高剂量组小鼠的常压缺氧存活时间显著延长(P＜0.05),其他实验组虽差异不显著(P＞0.05),但缺氧存活时间也有所延长;在亚硝酸钠中毒存活实验中,GPE组的小鼠存活时间显著长于其他实验组(P＜0.05),且与对照组差异达到显著水平(P＜0.01),GPPⅠ组和GPPⅡ四个实验组小鼠存活时间也比对照组显著延长(P＜0.05),且实验各组间差异水平显著(P＜0.05);在急性脑缺血性缺氧实验中,GPPⅡ四个实验组小鼠的脑缺氧存活时间均显著长于对照组和GPPⅠ组(P＜0.01),且其高剂量组与GPE组相比也达到极显著水平(P＜0.01),GPPⅡ的另三个剂量组与GPPⅠ组相比差异极显著(P＜0.01),且各实验组间差异也极显著(P＜0.01)。以上结果可以的看出,GPPⅡ具有增强小鼠机体缺氧耐受力的作用,且增强作用明显优于GPPⅠ和GPE。综合以上研究结果,认为GPE、GPPⅠ和GPPⅡ三种羊胎盘制剂对动物机体均无毒副作用和致突变作用,且具有提高机体免疫力和缺氧耐受力的作用。在本实验设计的配方中,GPPⅡ的效果最佳。更多还原

【Abstract】 The goat placenta extracts were prepared by technological procedure and produced two kinds of goat placenta preparations(GPPⅠand GPPⅡ)which were respectively prepared by mixing the extracts of goat placentas with other kinds of supplementary materials according to two formulas designed.The amount of goat placenta extract in each capsule(net weight 500mg) of GPPⅠor GPPⅡis equivalent to 30g fresh placenta.The test of toxicity was based on the effects on immune function and hypoxia tolerance of the extracts and two kinds of different capsules of goat placenta were conducted in mice.These tests included acute toxicity test,30 days’ treatment test in healthy condition,testicle chromosome aberration test,and myeloid cell micronucleus test. The immune function tests included lymphocyte transformation induced by concanavalin A and NK cytoactive of mice.The hypoxia tolerance tests included normal pressure hypoxia, sodium nitrite intoxation and acute cerebral ischemia.The mice in every group for each test for GPE,GPPⅠand GPPⅡwere fed one dosage to determine the optimal prescription of GPPⅡon the base of the former study. The mice were classed into three experiment groups and one control group randomly in GPPⅡ,There are 5 to 20 mice in every group according to the schedule.These mice were fed by artificial intragastric administration.The results are as follows:1.The results of acute toxicity test of GPE,GPPⅠand GPPⅡshowed that all treated mice were in good healthiness and no treated mouse presented abnormality or death,2.After treatment for 30 days,the mice in all experiment groups presented normal physiobehaviors,no toxic or death cases and no significant differences between the groups in utilization rate of food and hematology and blood biochemical indicators(P＞0.05),no pathological changes in the organs which were pathos copy,and no significant differences between the absolute weight and relative weight in the majority organs(P＞0.05),but absolute weight and relative weight of the spleen in group of GPPⅡwas significantly different to the index of control group(P＜0.05).3.Ames test showed that colony did not cause double increase of revertants at all doses in every bacterial strains no matter add the active system S-9 or not,and the dose-response relationship was not found.The results of Ames tests showed that GPE、GPPⅠand GPPⅡhad no mutagenic action.4.In the experiment of chromosome aberration in testicle cells of the mice experiment, the fragment and interchange amounts in all experiment groups’ testicle chromosome and the chromosome(including euchromosome and sex chromosome) had the univalent number,the distortion cell number and the distortion factor had no significant differences compared with the negative control group(P＞0.05),but had significant differences from the positive control group treated CTX(P＜0.01).5.The micronuclear rates of the mice in all dosage groups had no significant differences from the negative control groups(P＞0.05),however,the micronuclear rates of the positive control group was significantly higher than the others(P＜0.01).6.The results of mouse blastisation of lymphocyte tests and NK cytoactive tests indicated that mice’s splenic lymphocyte reproductive ability was significantly strengthened in GPPⅠand GPPⅡat single and middle dosage groups(P＜0.05),mice’s splenic lymphocyte reproductive ability of GPPⅡwas low,middle dosage groups were most significantly(P＜0.01),but there were no significant difference in GPE group,and splenic lymphocyte reproductive ability of GPPⅡlow,high dosage groups were significantly different from the GPEⅠor GPE group(P＜0.05);NK cytoactive of GPPⅡsingle,low,middle dosage groups were significantly strengthened(P＜0.05),more over,the low dosage’s difference was the most significant,while the others’ were not significant(P＞0.05).7.The results of hypoxia tolerance tests indicated that the mice feed GPPⅡat single, middle,and high dosage groups’ survival time were longer than that of the control group(P＜0.05),the other groups’ survival time were not significantly longer.In the sodium nitrite toxic texts,the mice of GPE groups’ survival time were significantly longer than the others(P＜0.05),and was significantly longer than the control group(P＜0.01),the mice of four groups’ survival time in GPPⅠand GPPⅡgroups were significantly longer than the control group(P＜0.05),and the difference between groups were significant(P＜0.05);in Acute cerebral ischemia tests,the survival time of the mice of four groups in GPPⅡwere significantly longer than the control and GPPⅠ(P＜0.01),and the survival time of the mice of the high dosage group was significantly longer than GPE group(P＜0.01),and the difference between groups was also most significantly(P＜0.01).These results above indicated that GPPⅡcould strengthen the mice’s hypoxia tolerance,and the function of GPPⅡwas significantly superior to GPPⅠand GPE.To sum up,GPE、GPPⅠand GPPⅡhad innoculty to the body,and had obvious favorable effects on the abilities of immunity and hypoxia tolerance.And GPPⅡwas the best among them.更多还原