本文選題：卵巢儲備功能　切入點：免疫　出處：《南京中醫(yī)藥大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：通過觀察雷公藤多甙(GTW)致卵巢儲備功能下降(DOR)大鼠免疫細(xì)胞因子白細(xì)胞介素-2(IL-2)的水平和卵巢腫瘤壞死因子-α(TNF-α)的表達,進一步探討補腎活血方改善卵巢儲備功能的作用機制,為臨床治療DOR提供科學(xué)依據(jù)。 方法：選取SD雌性健康大鼠48只,根據(jù)體重隨機分為6組(n=8)：空白對照組(灌胃同體積的生理鹽水)、GTW模型組(GTW50mg/kg/d)、六味地黃湯組(GTW50mg+六味地黃湯7.50g生藥/kg/d)、補腎活血方低劑量組(GTW50mg+補腎活血方3.25g生藥/kg/d)、補腎活血方中劑量組(GTW50mg+補腎活血方7.50g生藥/kg/d)、補腎活血方高劑量組(GTW50mg+補腎活血方15.00g生藥/kg/d)。連續(xù)15天灌胃給藥,給藥結(jié)束后1-4日內(nèi)結(jié)合陰道細(xì)胞學(xué)涂片,將未出現(xiàn)明顯動情周期的大鼠乙醚麻醉,予股動脈取血后頸椎脫臼處死,并立刻摘取大鼠雙側(cè)卵巢。觀察各組大鼠動情周期、體重、卵巢質(zhì)量、卵巢系數(shù)；采用放射免疫法檢測大鼠血清中FSH、E2的變化；采用放射免疫法檢測大鼠血清中免疫細(xì)胞因子IL-2的變化；HE染色檢測大鼠卵巢組織形態(tài)學(xué)的改變；免疫組化法檢測大鼠卵巢組織TNF-α水平的表達。 結(jié)果：(1)模型組大鼠卵巢質(zhì)量及卵巢系數(shù)較空白對照組降低；模型組大鼠血清中FSH明顯高升,E2明顯下降,與空白對照組比較均有顯著差異(P值均為p0.001)；模型組大鼠血清中免疫細(xì)胞因子IL-2下降,與空白對照組比較均有顯著差異(p0.01)；模型組大鼠卵巢組織中各期卵泡數(shù)均明顯減少,卵泡間質(zhì)內(nèi)有不同程度的炎細(xì)胞浸潤及纖維化；模型大鼠TNF-α呈低表達與空白對照組有顯著差異(p0.05)。 (2)補腎活血方低、中、高劑量組與六味地黃湯組均可不同程度改善GTW大鼠的卵巢質(zhì)量及卵巢系數(shù)；均能明顯降低GTW大鼠血清FSH水平,與模型組比較均有顯著差異(p0.001)；補腎活血方中、高劑量組與六味地黃湯組均可升高血清E2水平,與模型組比較均有差異(P值分別為p0.01、p0.05、p0.01);補腎活血方中、高劑量組與六味地黃湯組均可升高血清中免疫細(xì)胞因子IL-2水平,與模型組比較均有差異(均為p0.05)；補腎活血方中、高劑量組卵泡顆粒細(xì)胞和卵母細(xì)胞中的TNF-α表達較模型組改善,存在統(tǒng)計學(xué)差異(P0.05),與空白組比較無明顯差異。 結(jié)論：(1)補腎活血方可降調(diào)GTW大鼠血清中FSH水平,上調(diào)血清中E2水平,表明補腎活血方可改善卵巢儲備功能,上調(diào)血清中免疫細(xì)胞因子IL-2,拮抗卵巢中卵泡的損傷,升高卵泡顆粒細(xì)胞和卵母細(xì)胞TNF-α的表達,可能是其防治DOR的又一作用機制之一。 (2)補腎活血方(補腎活血法)和六味地黃湯(補腎法)均能改善卵巢儲備功能,抑制卵泡的損傷,但補腎活血方在改善卵泡顆粒細(xì)胞和卵母細(xì)胞TNF-α的表達方面優(yōu)于六味地黃湯,與前期研究基礎(chǔ)一致,反映出補腎活血方可通過多方面作用機制來改善DOR。
[Abstract]:Objective: to observe the level of interleukin-2 (IL-2) and the expression of tumor necrosis factor- 偽 (TNF- 偽) in rat ovarian reserve dysfunction induced by GTW (Tripterygium wilfordii). To explore the mechanism of Bushen Huoxue recipe in improving ovarian reserve function and provide scientific basis for clinical treatment of DOR. Methods: 48 SD female healthy rats were selected. According to body weight, they were randomly divided into six groups: blank control group (fed with normal saline of the same volume and GTW model group), GTW 50 mg / kg / kg / d group, Liuwei Dihuang decoction group with GTW 50 mg / kg / kg / kg / d, low dose group with low dose of Bushen Huoxue recipe (GTW 50 mg / kg / kg / kg / d), low dose group with low dose of Bushen Huoxue recipe (GTW 50 mg / kg / kg / kg / d), and the group with tonifying kidney. GTW 50mg Bushen Huoxue prescription 7.50g crude drug / kg 路kg 路d ~ (-1 / d) in medium dose group and 15.00 g / kg / kg / d GTW _ (50mg) Bushen / Huoxue prescription / high dose group were administrated by stomach for 15 days. Within 1-4 days after administration, combined with vaginal cytological smear, the rats without obvious estrous cycle were anesthetized with ether, and were killed by cervical dislocations after femoral artery blood extraction, and bilateral ovaries were removed immediately. The estrus cycle and body weight of the rats in each group were observed. The changes of serum FSHE _ 2 in rats were detected by radioimmunoassay, the changes of immunocyte factor IL-2 in serum by radioimmunoassay and the morphological changes of ovary by HE staining. The expression of TNF- 偽 in rat ovary was detected by immunohistochemistry. Results the ovarian weight and ovarian coefficient in the model group were lower than those in the blank control group, and the serum FSH in the model group was significantly higher than that in the control group. Compared with the blank control group, there were significant differences (P < 0.05) between the model group and the blank control group, the decrease of serum immunocyte factor IL-2 in the model group and the significant difference between the model group and the blank control group, the number of follicles in the ovarian tissue of the model group were significantly decreased, and the number of follicles in the ovarian tissue of the model group was significantly lower than that of the blank control group. There were different degrees of inflammatory cell infiltration and fibrosis in the interstitial follicle, and the low expression of TNF- 偽 in the model rats was significantly different from that in the blank control group (p 0.05). (2) Bushen Huoxue recipe was low, medium and high dose groups and Liuwei Dihuang decoction group could improve ovary quality and ovarian coefficient of GTW rats in different degree, and could obviously reduce the level of serum FSH in GTW rats. There were significant differences between the model group and the model group (P 0.001), in the high dosage group and the Liuwei Dihuang decoction group, the serum E2 level was increased, and the difference P values were respectively p0.01 (P < 0.05) and 0.05% (P < 0.01), respectively, in the Bushen Huoxue recipe, and in the Bushen Huoxue recipe, the serum E _ 2 level was significantly higher in the high dose group than in the model group. The levels of IL-2 in serum of high dose group and Liuwei Dihuang decoction group were significantly higher than those of model group (all p0.05), and the expression of TNF- 偽 in follicular granulosa cells and oocytes in high dose group was better than that in model group (all p0.05), and the expression of TNF- 偽 in follicular granulosa cells and oocytes in high dose group was better than that in model group. There was statistical difference (P 0.05), and there was no significant difference between the control group and the control group. Conclusion (1) Bushen Huoxue decoction can decrease the level of FSH in serum and increase the level of E2 in serum of GTW rats, indicating that Bushen Huoxue decoction can improve the function of ovarian reserve, upregulate the immune cytokine IL-2 in serum, and antagonize the damage of follicle in ovary. The increased expression of TNF- 偽 in follicular granulosa cells and oocytes may be one of its mechanisms in the prevention and treatment of DOR. (2) both Bushen Huoxue recipe (Bushen Huoxue method) and Liuwei Dihuang decoction (Kidney tonifying method) can improve ovarian reserve function and inhibit follicle injury, but Bushen Huoxue recipe is superior to Liuwei Dihuang decoction in improving the expression of TNF- 偽 in follicular granulosa cells and oocytes. Consistent with the previous study, Bushen Huoxue decoction can improve DORs through various mechanisms.
【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R285.5

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