本文選題：NF-κB　切入點(diǎn)：二甲雙胍　出處：《福建醫(yī)科大學(xué)》2014年碩士論文

【摘要】：[目的]1、建立可靠的PCOS大鼠模型，并設(shè)計(jì)二甲雙胍的給藥干預(yù)，研究NF-κB、IL-6、TNF-a在正常組、PCOS組、二甲雙胍干預(yù)組大鼠的表達(dá)，及與終體質(zhì)量、T、LH、HOMA-IR、E2表達(dá)的相關(guān)性，了解NF-κB及炎癥因子在PCOS中的作用；2、研究二甲雙胍對(duì)NF-κB、IL-6、TNF-a及終體質(zhì)量、T、LH、HOMA-IR、E2表達(dá)的影響，為臨床治療提供實(shí)驗(yàn)依據(jù)；3、探討二甲雙胍對(duì)多囊卵巢大鼠的促排卵作用及可能的機(jī)制。 [方法]將32只雌性S/D大鼠隨機(jī)分為三組，正常對(duì)照組（A組，n=10）：自由攝取食物和水31d；PCOS組(B組，n=12)：來曲唑1mg/kg灌胃21d后予以2.6mL/kg生理鹽水灌胃10d；二甲雙胍干預(yù)組（C組，n=10）：來曲唑1mg/kg灌胃21d后予以6%的二甲雙胍2.6mL/kg灌胃10d。每日陰道涂片及稱重，測(cè)定各組大鼠血清E2、T、LH、FPG、FINS水平，通過HE染色觀察卵巢形態(tài)，通過免疫組化、熒光定量PCR方法監(jiān)測(cè)各組大鼠卵巢NF-κB p65蛋白及mRNA的表達(dá)，通過Elisa測(cè)定各組大鼠循環(huán)中IL-6、TNF-a的表達(dá)，通過RT-PCR測(cè)定各組大鼠卵巢IL-6、TNF-a mRNA的相對(duì)表達(dá)。 [結(jié)果] 1.用來曲唑1mg/kg灌胃共21天，模型組大鼠于造模第9天均失去動(dòng)情周期，卵巢病理多囊改變，并具有高雄激素血癥和胰島素抵抗的特點(diǎn)，建立近似于人類PCOS的動(dòng)物模型； 2. PCOS組大鼠血清TNF-a顯著高于正常對(duì)照組（P0.05），二甲雙胍干預(yù)組血清TNF-a較PCOS組顯著下降（P0.05）；血清IL-6水平在三組大鼠間無顯著性差異（P0.05）；PCOS大鼠二甲雙胍干預(yù)后血清INS、T、LH顯著下降； 3.各指標(biāo)之間進(jìn)行Pearson相關(guān)性分析，很多各指標(biāo)之間存在相關(guān)，形成惡性循環(huán)；在相關(guān)的基礎(chǔ)上進(jìn)行多元線性回歸分析：TNF-a和體質(zhì)量是引起胰島素抵抗的獨(dú)立影響因素,可能是胰島素抵抗的機(jī)制之一；T和IL6對(duì)體質(zhì)量具有顯著性影響； IL-6是除HOMA-IR外，對(duì)T的獨(dú)立影響因子； 4.在二甲雙胍藥物干預(yù)的第6天，二甲雙胍組大鼠均出現(xiàn)動(dòng)情周期，恢復(fù)排卵；而PCOS組持續(xù)處于動(dòng)情間期，提示無排卵，直到第10天12只中僅1只恢復(fù)動(dòng)情周期。二甲雙胍干預(yù)組大鼠的卵巢形態(tài)均恢復(fù)正常，而PCOS組大鼠的卵巢形態(tài)仍為多囊改變； 5. PCOS大鼠卵巢局部TNF-a、IL-6mRNA的表達(dá)較正常對(duì)照組增高（P0.05），二甲雙胍干預(yù)組大鼠卵巢局部炎癥因子mRNA的表達(dá)較PCOS組顯著減少（P0.05）； 6.卵巢組織免疫組化顯示：NF-κB P65均一致性的表達(dá)在卵巢顆粒細(xì)胞的細(xì)胞核，但各組之間的表達(dá)無明顯統(tǒng)計(jì)學(xué)差別（P0.05），熒光定量PCR檢測(cè)卵巢組織NF-κB P65mRNA的表達(dá)，各組之間未發(fā)現(xiàn)顯著性差別（P0.05）。 [結(jié)論] 1. S/D大鼠來曲唑建�？勺鳛檠芯縋COS的理想動(dòng)物模型； 2.本PCOS大鼠模型存在慢性低度炎癥，可能由于臨床異質(zhì)性，，主要反映在TNF-a水平的升高，二甲雙胍的干預(yù)顯示有益的作用； 3. TNF-a是引起PCOS胰島素抵抗的獨(dú)立影響因素， IL-6是PCOS體質(zhì)量的獨(dú)立影響因素，可用于預(yù)測(cè)發(fā)生的風(fēng)險(xiǎn)性； 4.二甲雙胍具有良好的促排卵作用，能恢復(fù)卵巢正常形態(tài)，卵巢局部炎癥因子的改變可能直接參與發(fā)病機(jī)制； 5.炎癥因子的變化并不是通過NF-κB介導(dǎo)，可能存在其他信號(hào)通路，或者與PCOS臨床表現(xiàn)的異質(zhì)性有關(guān)，具體機(jī)制有待進(jìn)一步研究。
[Abstract]:Objective to establish the PCOS rat model of]1, reliable, and the design of metformin administration intervention, IL-6 of NF- kappa B, TNF-a, in the normal group, PCOS group, the expression of the intervention group rats and metformin, and the final body weight, T, LH, HOMA-IR, correlation between the expression of E2, to understand the role of B and inflammation factor NF- K in PCOS; 2, to study the effects of metformin on NF- kappa B, IL-6, TNF-a and final body mass, T, LH, HOMA-IR, E2 expression, and provide experimental basis for clinical treatment; 3, to investigate the effect of metformin on ovulation of polycystic ovary in rats and its possible mechanism.
[Methods] 32 female S/D rats were randomly divided into three groups, normal control group (group A, n=10): free access to food and water 31d; PCOS group (group B, n=12): letrozole intragastric administration of 1mg/kg 21d to 2.6mL/kg saline 10d; metformin intervention group (C group, n=10): letrozole intragastric Administration of 1mg/kg 21d to 6% 2.6mL/kg after intragastric administration of metformin 10d. daily vaginal smears and weighing, determination of serum E2, the rats of T, LH, FPG, FINS, observation of ovarian morphology by HE staining, immunohistochemistry, fluorescence quantitative PCR method for monitoring the expression of rats ovaries NF- p65 kappa B protein and mRNA, the rats were measured by Elisa TNF-a in the IL-6 cycle, the expression of ovarian IL-6 rats were measured by RT-PCR, the relative expression of TNF-a mRNA.
[results]
1., for 21 days, the rats in the model group were treated with trazole 1mg/kg for 21 days. The rats in the model group lost estrous cycle on the ninth day of modeling, ovarian pathology and polycystic changes, and had the characteristics of Kaohsiung hormone and insulin resistance. A animal model similar to human PCOS was established.
The serum TNF-a in 2. PCOS group was significantly higher than that in the normal control group (P0.05). The serum TNF-a in metformin intervention group was significantly lower than that in the PCOS group (P0.05), and there was no significant difference in serum IL-6 level between the three groups (P0.05). After PCOS metformin treatment, the serum INS, T and PCOS decreased significantly.
Pearson correlation analysis was performed between the 3. indexes, there are correlation between many indexes form a vicious spiral; multiple linear regression analysis in related on the basis of TNF-a and body mass is caused by the independent influencing factors of insulin resistance may be one of the mechanisms of insulin resistance; T and IL6 had significant effect on body weight; IL-6 is in addition to HOMA-IR, the independent impact factor of T;
4. in the metformin drug intervention for sixth days, the metformin group was observed in rat estrous cycle, ovulation; and PCOS Group continued in diestrus, suggesting no ovulation, until the tenth day of 12 in only 1 recovery of estrous cycle. Ovarian morphology in rats intervention group metformin were return to normal, and ovarian morphology in PCOS group the rat is polycystic change;
5., the expression of TNF-a and IL-6mRNA in the ovary of PCOS rats was higher than that in the normal control group (P0.05). The expression of mRNA in the ovary of the metformin treated group was significantly lower than that in the PCOS group (P0.05).
6. ovarian tissue immunohistochemistry showed that: NF- kappa B P65 were similar in the granulosa cell nuclei of expression, but the expression was no significant difference between the groups (P0.05), the expression of fluorescence quantitative PCR detection of ovarian tissue NF- kappa B P65mRNA, significant differences were found between the groups (P0.05).
[Conclusion]
1. S/D rat model of letrozole can be used as an ideal animal model for the study of PCOS.
2., there is chronic low grade inflammation in the PCOS rat model. It may be due to the clinical heterogeneity, which is mainly reflected in the increase of TNF-a level. Metformin intervention has a beneficial effect.
3. TNF-a is an independent factor causing PCOS insulin resistance, and IL-6 is an independent factor in the mass of PCOS body, which can be used to predict the risk of occurrence.
4. metformin has good ovulation stimulating effect and can restore the normal morphology of the ovary. The changes of local inflammatory factors of the ovary may be directly involved in the pathogenesis.
5., the change of inflammatory factors is not mediated by NF- kappa B. There may be other signaling pathways, or are related to the heterogeneity of PCOS clinical manifestations. The specific mechanism needs further study.

【學(xué)位授予單位】：福建醫(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

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 翟軍;孫瑩璞;;二甲雙胍對(duì)PCOS患者血清TNF-α水平及內(nèi)分泌代謝的影響[J];鄭州大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2011年02期

2 李巍巍;侯麗輝;郝松莉;李勤;吳效科;;多囊卵巢綜合征動(dòng)物模型構(gòu)建與評(píng)價(jià)[J];世界中西醫(yī)結(jié)合雜志;2011年10期

3 張娟;朱桂金;王昕榮;徐蓓;胡琳莉;;Apoptosis and Expression of Protein TRAIL in Granulosa Cells of Rats with Polycystic Ovarian Syndrome[J];Journal of Huazhong University of Science and Technology(Medical Sciences);2007年03期

4 章常華;胡憲紅;張忠偉;劉玉暉;余日躍;徐國(guó)良;;腫瘤壞死因子-α誘導(dǎo)3T3-L1脂肪細(xì)胞胰島素抵抗模型的建立[J];中國(guó)實(shí)驗(yàn)方劑學(xué)雜志;2012年18期

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