复方苦参凝胶杀精避孕及抗炎的实验研究

发布时间：2018-03-05 17:00

本文选题：苦参　切入点：白头翁　出处：《华中科技大学》2014年博士论文　论文类型：学位论文

【摘要】：第一部分不同配比苦参白头翁乙醇提取物体外杀精的实验研究 [目的]研究不同配比苦参白头翁乙醇提取物的体外杀精能力,并探讨其杀精原理。 [方法] 1.收集15例正常男性的精液标本,经密度梯度离心法处理。浓度为0.5mg/mL的9个不同配比苦参白头翁乙醇提取物药液作用精液标本20s、2min、4min后,用计算机辅助精子分析系统(computer-assisted sperm analysis, CAS A)来评价药液作用后其对精子活力(A+B级)及精子各种运动参数的影响,包括精子曲线运动速度(curvilinear velocity, VCL)、直线运动速度(straight-line velocity, VSL)、平均路径速度(average path velocity,VAP)、精子头侧摆幅度(amplitude of lateral head displacement, ALH)、摆动性(wobble, WOB)及平均移动角度(mean angular displacement, MAD)等； 2.使用标本同上,分别检测9种不同配比的苦参白头翁乙醇提取物具有瞬间(20s)杀精能力时的最低有效浓度； 3.收集3例正常男性的精液标本用于电镜观察。通过对上述两个步骤的分析找出体外杀精能力最佳的苦参白头翁配比,对其作用后的精液标本进行透射电镜观察,观察精子形态学的变化。 [结果] 1.以3：1配比的苦参白头翁乙醇提取物,浓度为0.5mg/mL时能明显抑制精子的活力,并且对精子运动参数VCL、VSL、VAP、ALH、WOB及MAD等指标均产生了明显的抑制作用,与对照组比较有统计学意义； 2.9个不同配比的乙醇提取物中以3：1配比时,其具有瞬间杀精能力时的最低有效浓度最低,为3.5mg/mL; 3.以3：1配比的乙醇提取物作用精液标本后,通过电镜观察发现其精子细胞膜及顶体被破坏,而对照组精子未见明显异常。 [结论]苦参白头翁3：1配比时,其乙醇提取物具有最佳的体外杀精能力,且其杀精原理可能与其破坏了精子细胞膜及顶体有关。第二部分复方苦参凝胶节育的体内实验研究 [目的]研究复方苦参凝胶体内节育的效果并进行阴道刺激性实验。 [方法] 1.8周龄雌性SD大鼠60只,随机分为生理盐水组、空白凝胶组、复方苦参凝胶低(0.05g生药/g)、中(0.10g生药/g)、高剂量组(0.15g生药/g)以及壬苯醇醚组(阳性对照)。每晚雌鼠阴道内各给予相应药物200μL,然后与雄鼠按照1：1合笼进行交配,在交配成功后的第12天剖腹观察雌鼠孕株数,并计算避孕率； 2.8周龄雌性SD大鼠15只,随机分为生理盐水组、空白凝胶组及复方苦参凝胶高剂量组。每组每晚阴道内给予相应药物各200μL,连续2周,末次给药24h后处死大鼠,取出阴道组织,观察大鼠阴道有无充血、红肿,并进一步行病理组织学检查。 [结果] 1.与生理盐水组比较,复方苦参凝胶高剂量(0.15g生药/g)组能显著抑制雌鼠的孕株数(0.00±0.00vs11.00±：2.00,P0.05)；而与阳性对照组比较无统计学意义(0.00±0.00vs0.00±0.00,P0.05),复方苦参凝胶高剂量组避孕率为100%； 2.雌鼠阴道连续给药2周后,各组大鼠阴道肉眼观均未见充血红肿,阴道病理组织检查示结构正常无炎症细胞浸润。 [结论]复方苦参凝胶有较好的节育作用,并且对阴道局部无不良反应。第三部分复方苦参凝胶抗炎的体内实验研究 [目的]研究复方苦参凝胶体内抗炎的效果。 [方法]采用小鼠耳廓双面均匀涂抹二甲苯的致炎模型,将60只昆明种小鼠随机分为6组：生理盐水组、空白凝胶组、复方苦参凝胶低(0.05g生药/g)、中(0.10g生药/g)、高剂量组(0.15g生药/g)以及阳性对照组(丙酸氯倍他索乳膏)。各组于每只小鼠右耳双侧均匀涂抹相应药物各50μL,每天1次,连续4天,末次给药后1h,于右耳双侧均匀涂抹100%二甲苯50μL,然后计算小鼠单侧耳的肿胀度及肿胀抑制率。 [结果]小鼠耳廓二甲苯致炎模型中,与生理盐水组比较,高剂量组能显著抑制其肿胀程度(10.17+2.56vs21.32±±3.17,P0.01),与阳性对照组比较无明显统计学差异(10.17±2.56vs8.53±1.89,P0.05),其肿胀抑制率为52.3%。 [结论]复方苦参凝胶有较好的抗炎作用。
[Abstract]:The first part of the study of different proportions of Baitouweng Kushen ethanol extracts of spermicide
[Objective] the effects of different ratio of ethanol extract of Sophora Pulsatilla chinensis in vitro spermicidal ability, and to explore its spermicidal principle.
[method]
1. collect 15 normal semen samples by density gradient centrifugation method. The concentration of 0.5mg/mL 9 with different ratio of ethanol extract of Sophora Pulsatillae liquid medicine effect of semen samples were 20s, 2min, 4min, using computer assisted sperm analysis system (computer-assisted sperm analysis, CAS A) to evaluate the effect of the drug on sperm motility (A+B) and sperm motion parameters, including sperm velocity curve (curvilinear velocity, VCL), straight line velocity (straight-line velocity, VSL), average path velocity (average path, velocity, VAP), the sperm head side swing (amplitude of lateral head displacement, ALH), (wobble, wobble WOB (mean) and the average angle of angular displacement, mobile MAD) etc.;
2. specimens were detected as above, 9 different kinds of Baitouweng Kushen ethanol extract (20s) has the lowest effective concentration of the spermicidal ability;
3. semen specimens were collected from 3 cases of normal male for electron microscopy. The in vitro spermicidal ability best found through the analysis of the ratio of Baitouweng Kushen of the above two steps, observed by transmission electron microscopy of the semen samples after, to observe the changes of sperm morphology.
[results]
1. to 3:1 ratio of ethanol extract of Baitouweng Kushen, when the concentration of 0.5mg/mL can inhibit the activity of sperm, and sperm motility parameters VCL, VSL, VAP, ALH, WOB and MAD were inhibited significantly, compared with the control group;
The minimum effective concentration of 2.9 ethanol extracts with 2.9 different ratios was the lowest when it had the ability of instantaneous spermatogenic ability, which was 3.5mg/mL.
3., after ethanol extracts from 3:1 were used to semen samples, the sperm cell membrane and acrosome were destroyed by electron microscope, while the sperm in control group was not abnormal.
[Conclusion] Baitouweng Kushen 3:1 ratio, the ethanol extract has the best in vitro spermicidal ability and its spermicidal principle may be related to damage the sperm cell membrane and acrosome.
Experimental study on the second part of compound Sophora flavescens gel in vivo
[Objective] to study the effect of the compound Kushen gel in vivo and to carry out the vaginal irritation test.
[method]
1.8 week old female SD 60 rats were randomly divided into saline group, blank gel group, matrine gel low (0.05g crude drug /g), medium (0.10g crude drug /g), high dose (0.15g crude drug /g) and nonoxinol group (positive control). The corresponding drugs 200 L in the night female rats in the vagina, and then according to 1:1 male rats mated female rats were mated to observe the pregnancy number in mating success on the twelfth day after laparotomy, and calculate the rate of contraception;
2.8 week old female SD 15 rats were randomly divided into saline group, blank gel group and high dose group of compound matrine gel. Each night the vagina with corresponding drugs for 200 L, for 2 weeks, 24h after the last administration the rats were sacrificed, remove the vaginal tissue, observe the rat vagina without full of blood, swelling, and further histopathological examination.
[results]
1. compared with normal saline group, high dose matrine gel (0.15g crude drug /g) group can significantly inhibit the number of pregnant female rats (0 + 0.00vs11.00 + 2, P0.05); and compared with the positive control group had no statistical significance (0 + 0.00vs0.00 + 0, P0.05), the high dose group of compound Matrine gel contraceptive the rate is 100%;
After 2 weeks of continuous vaginal administration of 2. female rats, no hyperemia and swelling of the vagina were found in all rats. The pathological examination of the vagina showed that the structure was normal and no inflammatory cells were infiltrated.
[Conclusion] compound Sophora flavescens gel has good effect on birth control and has no adverse reaction to the vagina.
The third part
Experimental study on the anti-inflammatory effect of compound Sophora flavescens gel in vivo
[Objective] to study the anti inflammatory effect of compound Sophora flavescens gel in vivo.
[method] using the mouse ear inflammation model induced by xylene double face evenly, 60 Kunming mice were randomly divided into 6 groups: normal saline group, blank gel group, matrine gel low (0.05g crude drug /g), medium (0.10g crude drug /g), high dose (0.15g crude drug /g) and positive control group (Clobetasonl Propionate Cream). Each group in each mouse ear bilateral evenly corresponding drugs 50 L, 1 times a day, for 4 days after the last injection of 1H, on the right ear bilateral evenly 100% xylene 50 L, and then calculate the mouse single ear swelling and swelling inhibition rate.
[results] the mouse ear inflammation model induced by xylene, compared with the normal saline group, high dose group could significantly inhibit the swelling degree (10.17+2.56vs21.32 + + 3.17, P0.01), there was no significant difference between the positive control group (10.17 + 2.56vs8.53 + 1.89, P0.05), the inhibition rate was 52.3%.
[Conclusion] compound Sophora flavescens gel has good anti-inflammatory effect.

【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R698

【参考文献】