本文選題：多囊卵巢綜合征 + 大鼠�。� 參考：《山西醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的：多囊卵巢綜合征(PCOS)是常見的女性排卵障礙性疾病，臨床表現(xiàn)為月經(jīng)稀發(fā)、不孕、多毛、肥胖等。但迄今病因未明，臨床治療效果欠佳。 Cx43是顆粒細(xì)胞膜表面的連接蛋白。GATA-4是一種轉(zhuǎn)錄調(diào)節(jié)因子。June等動(dòng)物研究發(fā)現(xiàn)Cx43與顆粒細(xì)胞的正常發(fā)育有關(guān)。White等發(fā)現(xiàn)GATA-4與顆粒細(xì)胞的增殖分化密切相關(guān)。二者均與卵泡的正常發(fā)育有關(guān)。探討Cx43和GATA-4在PCOS模型大鼠卵巢的表達(dá)及復(fù)方甘芍貼劑干預(yù)作用機(jī)制。本課題組前期研究結(jié)果證實(shí)，復(fù)方甘芍貼劑治療PCOS模型大鼠，在血清性激素水平、光鏡和電鏡下的組織學(xué)變化均與目前臨床PCOS推薦治療藥物達(dá)英-35的效果類似。本研究擬通過Cx43和GATA-4在研究組與對(duì)照組中的變化來探討復(fù)方甘芍貼劑可能的治療機(jī)制。 方法：將48只6周齡的SD雌性大鼠,首先隨機(jī)分為兩組,正常對(duì)照組(12只)和實(shí)驗(yàn)對(duì)照組(36只),正常對(duì)照組同期灌服1%CMC(1ml/100g),1次/d,連續(xù)3周。實(shí)驗(yàn)對(duì)照組用來曲唑建立多囊卵巢綜合征（PCOS）大鼠模型,采用化學(xué)發(fā)光法檢測(cè)大鼠血清性激素水平,符合PCOS模型的大鼠進(jìn)入下一步研究,將36只大鼠隨機(jī)分為PCOS模型組（對(duì)照）、達(dá)英-35（炔雌醇環(huán)丙孕酮）對(duì)照組和外用復(fù)方甘芍貼劑研究組。達(dá)英-35（炔雌醇環(huán)丙孕酮）組為自9周齡起，按人與大鼠公斤體重折算系數(shù)給藥，將藥物溶于1%CMC(1ml/100g)給藥，1次/d，連續(xù)3周；復(fù)方甘芍外用貼劑組64d齡時(shí)，脫毛后并給予貼劑，1貼/d，并且同期灌服1%CMC(1ml/100g),1次/d，連續(xù)3周；PCOS模型組和正常對(duì)照組大鼠同期灌服同體積1%CMC (1ml/100g)，1次/d，共3周。其中模型組和達(dá)英-35組因操作不當(dāng)，，灌死兩只，各剩下11只。剩余46只于84日齡取血清測(cè)睪酮(T),卵泡生成素(FSH),黃體生成素(LH),取雙側(cè)卵巢,左側(cè)置于10%的甲醛液中固定24小時(shí)，用免疫組織化學(xué)觀察Cx43和GATA-4在PCOS模型大鼠的表達(dá)情況;右側(cè)卵巢放于-70度冰箱凍存,采用Western blot方法觀察Cx43和GATA-4在實(shí)驗(yàn)大鼠卵巢顆粒細(xì)胞中的蛋白表達(dá)情況。采用SPSS17.0統(tǒng)計(jì)軟件進(jìn)行分析;數(shù)據(jù)均以均數(shù)±標(biāo)準(zhǔn)差表示,多樣本間比較采用單因素方差分析，兩兩比較用LSD-t檢驗(yàn)兩組間均數(shù)間相關(guān)性分析采用Pearson相關(guān)。以P＜0.05為差異具有統(tǒng)計(jì)學(xué)意義。結(jié)果： 1.與正常對(duì)照組比較,正常對(duì)照組大鼠血清FSH水平為（2.41±0.23）IU/L，LH水平為（8.11±0.85）IU/L，T水平為（2.3±0.96）nmol/L,36只來曲唑誘導(dǎo)的多囊卵巢綜合征模型大鼠血清FSH、LH、T水平分別為FSH（2.95±0.21）IU/L，LH（8.72±0.99）IU/L，T（3.18±1.46）nmol/L，提示造模成功。 2.免疫組織化學(xué)染色:（1）與正常組大鼠卵巢Cx43灰度值(149.59±9.25)比較, PCOS模型組大鼠卵巢Cx43灰度值(98.19±6.51)，差異具有顯著性（P＜0.05）；與PCOS模型組大鼠卵巢Cx43灰度值(98.19±6.51)比較，復(fù)方甘芍貼劑組大鼠卵巢顆粒細(xì)胞膜中Cx43棕黃色顆粒陽性表達(dá)灰度值（144.28±4.72），差異具有顯著性（P＜0.05）；與達(dá)英-35（炔雌醇環(huán)丙孕酮）對(duì)照組（143.79±9.81）相比，復(fù)方甘芍貼劑組大鼠卵巢顆粒細(xì)胞膜中Cx43棕黃色顆粒陽性表達(dá)灰度值（144.29±4.72），沒有顯著性差異（P0.05）。（2）與正常組大鼠卵巢灰度值GATA-4(129.37±9.99)比較, PCOS模型組大鼠卵巢GATA-4灰度值(162.91±9.38)，差異具有顯著性（P＜0.05）；與PCOS模型組大鼠卵巢GATA-4灰度值(162.91±9.38)比較，復(fù)方甘芍貼劑組大鼠卵巢GATA-4棕黃色顆粒陽性表達(dá)灰度值（115.13±7.04），差異具有顯著性（P＜0.05）；與達(dá)英-35（炔雌醇環(huán)丙孕酮）治療組（112.37±7.90），復(fù)方甘芍貼劑組大鼠卵巢GATA-4棕黃色顆粒陽性表達(dá)灰度值（115.13±7.04），沒有顯著性差異（P0.05）。 3.采用Western blot方法檢測(cè)結(jié)果:（1）與正常組大鼠卵巢（0.86±0.10）比較，PCOS模型組大鼠卵巢Cx43蛋白表達(dá)（0.66±0.15），差異具有顯著性（P＜0.05）；與PCOS模型組大鼠卵巢Cx43(0.66±0.15)比較，復(fù)方甘芍貼劑組大鼠卵巢（0.86±0.10），差異具有顯著性（P＜0.05）；與達(dá)英-35（炔雌醇環(huán)丙孕酮）治療組（0.88±0.12），復(fù)方甘芍貼劑組大鼠卵巢GATA-4蛋白表達(dá)（0.86±0.10），沒有顯著性差異（P0.05）。（2）與正常組大鼠卵巢GATA-4（0.90±0.10）比較，PCOS模型組大鼠卵巢GATA-4蛋白表達(dá)（1.64±0.16），差異具有顯著性（P＜0.05）；與PCOS模型組大鼠卵巢GATA-4比較，復(fù)方甘芍貼劑組大鼠卵巢GATA-4（1.00±0.09），差異具有顯著性（P＜0.05）；與達(dá)英-35（炔雌醇環(huán)丙孕酮）治療組（1.15±0.32）比較，復(fù)方甘芍貼劑組大鼠卵巢GATA-4蛋白表達(dá)（1.00±0.09），沒有顯著性差異（P0.05）。 結(jié)論：1. PCOS模型大鼠卵巢顆粒細(xì)胞Cx43表達(dá)減少, GATA-4表達(dá)增加與多囊卵巢綜合征的發(fā)病可能有關(guān).2.復(fù)方甘芍貼劑治療后, PCOS模型大鼠卵巢顆粒細(xì)胞Cx43表達(dá)增加,GATA-4表達(dá)下降,提示該貼劑可能通過調(diào)節(jié)Cx43與GATA-4水平,改善卵泡的發(fā)育,抑制其凋亡而發(fā)揮治療作用。
[Abstract]:Objective: polycystic ovary syndrome (PCOS) is a common female ovulation disorder. The clinical manifestations of polycystic ovary syndrome are dilute menstruation, infertility, hairy, obesity and so on. But so far the etiology is not clear and the effect of clinical treatment is not good. Cx43 is a connexin.GATA-4 on the surface of granular cell membrane, a transcription regulator.June and other animal studies found Cx43 and granular cells The normal development related to.White found that GATA-4 was closely related to the proliferation and differentiation of granulosa cells. All two were related to the normal development of follicle. The expression of Cx43 and GATA-4 in the ovary of the rat model of PCOS and the intervention mechanism of compound Gump peony patch were discussed. The results of the previous study in this group confirmed that the compound Gump peony patch was used to treat PCOS model rats, The level of serum sex hormone, the histological changes under light and electron microscopy are similar to that of the current clinical PCOS recommended medications, Da Ying -35. This study intends to explore the possible therapeutic mechanism of compound Gump peony patch by the changes of Cx43 and GATA-4 in the study group and the control group.
Methods: 48 SD female rats of 6 weeks old were randomly divided into two groups, the normal control group (12 rats) and the experimental control group (36 rats). The normal control group was given 1%CMC (1ml/100g), 1 /d and 3 weeks for 3 weeks. The experimental control group was used to establish the rat model of polycystic ovary syndrome (PCOS), and the serum sex hormone was detected by chemiluminescence. 36 rats were randomly divided into PCOS model group (control), Da Ying -35 (estriol proprop progesterone) control group and foreign compound Gump peony patch group. The group of Da British -35 (estriol proprop) group was from 9 weeks of age, and the drug was dissolving in 1%CMC by human and rat kg body weight conversion coefficient. (2) 36 rats were randomly divided into the model group (control), the control group of Da Ying (estriol proprop) and the compound Gump peony patch group. 1ml/100g) administration, 1 times /d, for 3 weeks. When the compound Gump peony external use patch group was 64D, it was depilated and given a patch, 1 /d, and 1%CMC (1ml/100g) and 1 /d for 3 weeks for the same period; PCOS model group and normal control group were given the same volume 1%CMC (1ml/100g), 1 /d, for 3 weeks. The model group and Da Ying -35 group were irrigated because of improper operation. The remaining 11 were left 11. The remaining 46 were serum testosterone (T), follicular hormone (FSH), luteinizing hormone (LH), bilateral ovary and 24 hour in the 10% formaldehyde solution. The expression of Cx43 and GATA-4 in PCOS model rats was observed by immunohistochemistry; the right ovary was stored in -70 degree refrigerator, and Western BL was used. Ot method was used to observe the protein expression of Cx43 and GATA-4 in the ovarian granulosa cells of experimental rats. The data were analyzed with SPSS17.0 statistical software. The data were expressed with mean standard deviation, and a single factor analysis of variance was used for the comparison. The correlation analysis between the two groups was compared with the LSD-t test, and the correlation analysis was correlated with Pearson. P < 0.05 was 0.05. The difference was statistically significant.
1. compared with the normal control group, the serum FSH level of the normal control group was (2.41 + 0.23) IU/L, the level of LH was (8.11 + 0.85) IU/L, the level of T was (2.3 + 0.96) nmol/L. The serum FSH, LH, T level of the rat model of polycystic ovary syndrome induced by letrozole was FSH (2.95 + 0.21) IU/L, LH (8.72 + 0.99), indicating the formation of the model. Work.
2. immunohistochemical staining: (1) compared with the Cx43 gray value (149.59 + 9.25) in the normal group, the Cx43 gray value of the ovary in the PCOS model group was significantly (98.19 + 6.51), and the difference was significant (P < 0.05). Compared with the Cx43 gray value (98.19 + 6.51) in the rat ovary of the PCOS model group (98.19 + 6.51), the Cx43 brown and yellow in the granulosa cell membrane of the compound Gump patch group was Cx43 brown and yellow. The gray value of the positive expression of color particles was (144.28 + 4.72), and the difference was significant (P < 0.05). Compared with the control group (143.79 + 9.81) of -35 (estriol proproparone) in the control group (143.79 + 9.81), the positive expression of Cx43 brown yellow granules in the granulosa cell membrane of the compound Gump patch group was (144.29 + 4.72), and there was no significant difference (P0.05). (2) and the normal group The gray value of rat ovaries was GATA-4 (129.37 + 9.99). The GATA-4 gray value of the ovary in the PCOS model group was (162.91 + 9.38), and the difference was significant (P < 0.05). Compared with the GATA-4 gray value (162.91 + 9.38) of the rat ovary of the PCOS model group (162.91 + 9.38), the positive expression of the GATA-4 brown yellow granules in the ovary of the compound Gump patch group was (115.13 + 7.04), and the difference was different. It was significant (P < 0.05); with the treatment group (112.37 + 7.90) of Da Ying -35 (estriol propropyl progesterone), the positive expression of GATA-4 brown yellow granules in the ovary of the compound Gump peony patch group was (115.13 + 7.04), and there was no significant difference (P0.05).
3. Western blot method was used to detect the results of (1) compared with the normal group of rat ovary (0.86 + 0.10), the expression of Cx43 protein in the ovary of the PCOS model group was (0.66 + 0.15), the difference was significant (P < 0.05). Compared with the Cx43 (0.66 + 0.15) of the rat ovary (0.66 + 0.15) in the model group, the difference was significant (P < 0) (P < 0) (P < 0). 5): with the treatment group (0.88 + 0.12) of -35 (0.88 + 0.12), there was no significant difference in the expression of GATA-4 protein (P0.05). (2) compared with the normal group of rat ovary (0.90 + 0.10), the expression of GATA-4 protein in the ovary of the PCOS model group was (1.64 + 0.16), and the difference was significant (P < 0.05). Compared with the rat ovarian GATA-4 in the PCOS model group, the ovary was GATA-4 (1 + 0.09) in the compound Gump patch group (P < 0.05). Compared with the treatment group (1.15 + 0.32) of Da Ying -35 (1.15 + 0.32), the expression of GATA-4 egg white in the ovary of the compound Gump patch group was (1 + 0.09), and there was no significant difference (P0.05).
Conclusion: 1. PCOS model rat ovarian granulosa cells Cx43 expression decreased, GATA-4 expression increased and the pathogenesis of polycystic ovary syndrome may be related to the.2. compound glycyrrhizin patch treatment, PCOS model rat ovarian granulosa cells Cx43 expression increased, GATA-4 expression decreased, suggesting that the patch can improve the follicle by regulating the level of Cx43 and GATA-4. Development, inhibition of its apoptosis and play a therapeutic role.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R711.75

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉茂如;鎘生產(chǎn)時(shí)混合性粉塵致病作用的評(píng)價(jià)[J];中國(guó)職業(yè)醫(yī)學(xué);1982年05期

2 王|蘭;芳香族烴類經(jīng)皮膚的滲透量[J];國(guó)外醫(yī)學(xué).衛(wèi)生學(xué)分冊(cè);1982年01期

3 張寅恭;新殺蟲劑Пиримор的衛(wèi)生毒理學(xué)[J];國(guó)外醫(yī)學(xué).衛(wèi)生學(xué)分冊(cè);1982年01期

4 蔡海英,張金生,崔小邢,鄭國(guó)墚,黃孔威;國(guó)產(chǎn)維甲酸衍生物對(duì)大鼠食管上皮癌變的抑制作用[J];癌癥;1983年04期

5 高強(qiáng);大鼠直腸給心得安以避免首過消除效應(yīng)[J];國(guó)外醫(yī)學(xué).藥學(xué)分冊(cè);1983年01期

6 Seigo SHUMIYA;Sumi NAGASE;蔡華琬;;大鼠(Rattus Norvegicus)無白蛋白基因(Aualbumia)和蓋帽基因(Hooded)的連鎖[J];實(shí)驗(yàn)動(dòng)物與比較醫(yī)學(xué);1983年01期

7 陳國(guó)志,黃淑英,王廣義,劉志紅;豚鼠和大鼠小腸移行性肌電復(fù)合波(MMC)的觀察[J];基礎(chǔ)醫(yī)學(xué)與臨床;1984年03期

8 橫路謙次郎;不同類型輻射照射復(fù)合催乳素誘發(fā)大鼠乳腺腫瘤的效果[J];輻射防護(hù);1984年03期

9 溫志永;;幼鼠夜間PR 峰的出現(xiàn)對(duì)孕酮的依賴作用[J];國(guó)外醫(yī)學(xué).婦產(chǎn)科學(xué)分冊(cè);1989年05期

10 趙學(xué)增;鄭富穩(wěn);夏燁;;普萘洛爾對(duì)雌性大鼠生殖機(jī)能的影響[J];河北醫(yī)科大學(xué)學(xué)報(bào);1989年03期

相關(guān)會(huì)議論文 前10條

1 程井軍;吳其愷;彭銳;孫國(guó)杰;;電針對(duì)非酒精性脂肪性肝炎大鼠肝組織氧化及抗氧化狀態(tài)的影響[A];中華中醫(yī)藥學(xué)會(huì)第十三屆內(nèi)科肝膽病學(xué)術(shù)會(huì)議論文匯編[C];2008年

2 楊s

本文編號(hào)：1911676