本文選題：苦參　切入點：白頭翁　出處：《華中科技大學(xué)》2014年博士論文　論文類型：學(xué)位論文

【摘要】：第一部分不同配比苦參白頭翁乙醇提取物體外殺精的實驗研究 [目的]研究不同配比苦參白頭翁乙醇提取物的體外殺精能力,并探討其殺精原理。 [方法] 1.收集15例正常男性的精液標(biāo)本,經(jīng)密度梯度離心法處理。濃度為0.5mg/mL的9個不同配比苦參白頭翁乙醇提取物藥液作用精液標(biāo)本20s、2min、4min后,用計算機(jī)輔助精子分析系統(tǒng)(computer-assisted sperm analysis, CAS A)來評價藥液作用后其對精子活力(A+B級)及精子各種運(yùn)動參數(shù)的影響,包括精子曲線運(yùn)動速度(curvilinear velocity, VCL)、直線運(yùn)動速度(straight-line velocity, VSL)、平均路徑速度(average path velocity,VAP)、精子頭側(cè)擺幅度(amplitude of lateral head displacement, ALH)、擺動性(wobble, WOB)及平均移動角度(mean angular displacement, MAD)等； 2.使用標(biāo)本同上,分別檢測9種不同配比的苦參白頭翁乙醇提取物具有瞬間(20s)殺精能力時的最低有效濃度； 3.收集3例正常男性的精液標(biāo)本用于電鏡觀察。通過對上述兩個步驟的分析找出體外殺精能力最佳的苦參白頭翁配比,對其作用后的精液標(biāo)本進(jìn)行透射電鏡觀察,觀察精子形態(tài)學(xué)的變化。 [結(jié)果] 1.以3：1配比的苦參白頭翁乙醇提取物,濃度為0.5mg/mL時能明顯抑制精子的活力,并且對精子運(yùn)動參數(shù)VCL、VSL、VAP、ALH、WOB及MAD等指標(biāo)均產(chǎn)生了明顯的抑制作用,與對照組比較有統(tǒng)計學(xué)意義； 2.9個不同配比的乙醇提取物中以3：1配比時,其具有瞬間殺精能力時的最低有效濃度最低,為3.5mg/mL; 3.以3：1配比的乙醇提取物作用精液標(biāo)本后,通過電鏡觀察發(fā)現(xiàn)其精子細(xì)胞膜及頂體被破壞,而對照組精子未見明顯異常。 [結(jié)論]苦參白頭翁3：1配比時,其乙醇提取物具有最佳的體外殺精能力,且其殺精原理可能與其破壞了精子細(xì)胞膜及頂體有關(guān)。 第二部分復(fù)方苦參凝膠節(jié)育的體內(nèi)實驗研究 [目的]研究復(fù)方苦參凝膠體內(nèi)節(jié)育的效果并進(jìn)行陰道刺激性實驗。 [方法] 1.8周齡雌性SD大鼠60只,隨機(jī)分為生理鹽水組、空白凝膠組、復(fù)方苦參凝膠低(0.05g生藥/g)、中(0.10g生藥/g)、高劑量組(0.15g生藥/g)以及壬苯醇醚組(陽性對照)。每晚雌鼠陰道內(nèi)各給予相應(yīng)藥物200μL,然后與雄鼠按照1：1合籠進(jìn)行交配,在交配成功后的第12天剖腹觀察雌鼠孕株數(shù),并計算避孕率； 2.8周齡雌性SD大鼠15只,隨機(jī)分為生理鹽水組、空白凝膠組及復(fù)方苦參凝膠高劑量組。每組每晚陰道內(nèi)給予相應(yīng)藥物各200μL,連續(xù)2周,末次給藥24h后處死大鼠,取出陰道組織,觀察大鼠陰道有無充血、紅腫,并進(jìn)一步行病理組織學(xué)檢查。 [結(jié)果] 1.與生理鹽水組比較,復(fù)方苦參凝膠高劑量(0.15g生藥/g)組能顯著抑制雌鼠的孕株數(shù)(0.00±0.00vs11.00±：2.00,P0.05)；而與陽性對照組比較無統(tǒng)計學(xué)意義(0.00±0.00vs0.00±0.00,P0.05),復(fù)方苦參凝膠高劑量組避孕率為100%； 2.雌鼠陰道連續(xù)給藥2周后,各組大鼠陰道肉眼觀均未見充血紅腫,陰道病理組織檢查示結(jié)構(gòu)正常無炎癥細(xì)胞浸潤。 [結(jié)論]復(fù)方苦參凝膠有較好的節(jié)育作用,并且對陰道局部無不良反應(yīng)。 第三部分 復(fù)方苦參凝膠抗炎的體內(nèi)實驗研究 [目的]研究復(fù)方苦參凝膠體內(nèi)抗炎的效果。 [方法]采用小鼠耳廓雙面均勻涂抹二甲苯的致炎模型,將60只昆明種小鼠隨機(jī)分為6組：生理鹽水組、空白凝膠組、復(fù)方苦參凝膠低(0.05g生藥/g)、中(0.10g生藥/g)、高劑量組(0.15g生藥/g)以及陽性對照組(丙酸氯倍他索乳膏)。各組于每只小鼠右耳雙側(cè)均勻涂抹相應(yīng)藥物各50μL,每天1次,連續(xù)4天,末次給藥后1h,于右耳雙側(cè)均勻涂抹100%二甲苯50μL,然后計算小鼠單側(cè)耳的腫脹度及腫脹抑制率。 [結(jié)果]小鼠耳廓二甲苯致炎模型中,與生理鹽水組比較,高劑量組能顯著抑制其腫脹程度(10.17+2.56vs21.32±±3.17,P0.01),與陽性對照組比較無明顯統(tǒng)計學(xué)差異(10.17±2.56vs8.53±1.89,P0.05),其腫脹抑制率為52.3%。 [結(jié)論]復(fù)方苦參凝膠有較好的抗炎作用。
[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.

【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R698

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