本文選題：子宮內(nèi)膜異位癥　切入點(diǎn)：動(dòng)物模型　出處：《浙江大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：背景 子宮內(nèi)膜異位癥(以下簡稱內(nèi)異癥)是指具有生長功能的子宮內(nèi)膜組織(腺體和間質(zhì))出現(xiàn)在子宮腔被覆蓋內(nèi)膜及宮體肌層以外的其他部位的婦科常見疾病,嚴(yán)重影響婦女身心健康.疼痛(包括痛經(jīng)、性交痛和慢性盆腔痛等)和不孕是內(nèi)異癥的主要臨床癥狀,其患病率在育齡婦女中達(dá)10-15%。雖然內(nèi)異癥發(fā)病率呈現(xiàn)增高趨勢,但其病因和發(fā)病機(jī)制以及相應(yīng)內(nèi)異癥疼痛的發(fā)生機(jī)制至今仍然不清。近年大量研究發(fā)現(xiàn),內(nèi)異癥病灶內(nèi)出血刺激導(dǎo)致無菌性炎癥,免疫炎癥以及病灶神經(jīng)異常生長導(dǎo)致中樞與外周神經(jīng)敏化可能是內(nèi)異癥疼痛的主要發(fā)病機(jī)制。 肥大細(xì)胞(Mast cells,MC)是免疫炎癥反應(yīng)中的重要細(xì)胞之一,一些學(xué)者研究發(fā)現(xiàn)MC脫顆粒與偏頭痛、間質(zhì)性膀胱炎、腸易激綜合癥、異位性皮炎等疾病的疼痛癥狀有明顯的相關(guān)性。在神經(jīng)病理性疼痛中,分布于神經(jīng)和神經(jīng)周圍組織的MC即可脫顆粒釋放類胰蛋白酶、腫瘤壞死因子(Tumor necrosis factor-α, TNF-α)、神經(jīng)肽、P物質(zhì)(Substance P, SP)、神經(jīng)生長因子(Nerve growth factor, NGF)、細(xì)胞因子以及神經(jīng)遞質(zhì)如乙酰膽堿等活性介質(zhì)導(dǎo)致神經(jīng)敏化,而敏化的神經(jīng)可釋放大量神經(jīng)遞質(zhì)反過來激活MC促使其脫顆粒釋放致痛物質(zhì),進(jìn)一步促進(jìn)神經(jīng)敏化,從而導(dǎo)致神經(jīng)病理性疼痛的持續(xù)狀態(tài)。研究發(fā)現(xiàn),內(nèi)異癥病灶存在MC數(shù)量增加以及其活性增強(qiáng),且病灶中MC與神經(jīng)纖維之間密切相關(guān)。內(nèi)異癥疼痛目前認(rèn)為也是一種內(nèi)臟和神經(jīng)病理性疼痛,因而MC在內(nèi)異癥疼痛發(fā)生機(jī)制中發(fā)揮作用。 研究表明,雌激素可結(jié)合MC膜上ER-a啟動(dòng)快速的鈣離子流,促進(jìn)人及小鼠MC活化脫顆粒釋放相應(yīng)的介質(zhì)。色甘酸鈉是MC膜穩(wěn)定劑,能抑制其釋放介質(zhì),其作用機(jī)制可能與MC78-kDa蛋白磷酸化有關(guān)。78-kDa蛋白由兩段與MC膜突蛋白同源的染色體組成,在細(xì)胞表面與細(xì)胞內(nèi)構(gòu)架之間通過調(diào)節(jié)性功能的連接而參與信號(hào)傳導(dǎo),一旦膜突蛋白在磷酸化／脫磷酸化作用下結(jié)構(gòu)發(fā)生改變,將導(dǎo)致細(xì)胞膜和細(xì)胞內(nèi)部結(jié)構(gòu)的位置重排,并可能使MC分泌顆粒。 基于以上研究,我們?cè)O(shè)計(jì)了本實(shí)驗(yàn),首先建立SD大鼠內(nèi)異癥模型,在此基礎(chǔ)上我們給予MC脫顆粒促進(jìn)劑(雌激素)及抑制劑(色甘酸鈉)干預(yù)后,觀察模型病灶大小變化以及與MC脫顆粒的關(guān)系,探討MC在內(nèi)異癥發(fā)病中的作用,為臨床內(nèi)異癥疼痛的治療提供新思路。 目的 探討肥大細(xì)胞在子宮內(nèi)膜異位癥發(fā)病機(jī)理中的作用。 材料和方法 取健康雌性未孕SD大鼠60只,采用自體子宮內(nèi)膜移植法建立大鼠腹壁內(nèi)異癥模型,分兩部分進(jìn)行：第一部分(每組8只)大鼠造模后即每天臀部肌肉注射不同劑量雌激素(高劑量組：造模時(shí)同時(shí)切除雙側(cè)卵巢后給予外源性雌激素200μg/kg,低劑量組：造模時(shí)同時(shí)切除雙側(cè)卵巢后給予外源性雌激素100μg/kg,單純?cè)炷＝M：造模時(shí)不切除雙側(cè)卵巢不給予外源性雌激素),兩周后觀察其對(duì)大鼠內(nèi)異癥病灶的大小,組織形態(tài)的影響,每組處死4只大鼠取材,剩余大鼠繼續(xù)注射相應(yīng)劑量雌激素兩周,取材,比較病灶MC總數(shù)及其脫顆粒數(shù)、類胰蛋白酶、NGF的表達(dá),血清TNF-α、雌二醇(Estradiol, E2)水平；第二部分(分別10只、10只、8只、8只)大鼠造模兩周后觀察大鼠內(nèi)異癥病灶的大小,隨后每天腹腔注射不同劑量色甘酸鈉(高劑量組20mg/kg,低劑量組10mg/kg,溶劑組,空白對(duì)照組),連續(xù)用藥兩周處死取材,檢測指標(biāo)同上。 統(tǒng)計(jì)方法 SPSS20.0軟件進(jìn)行分析,結(jié)果以X±S表示,進(jìn)行正態(tài)性和方差齊性檢驗(yàn),多組間數(shù)據(jù)比較用單因素方差分析,兩組間數(shù)據(jù)比較采用t檢驗(yàn),檢驗(yàn)水準(zhǔn)取0.05. 結(jié)果 1.雌激素干預(yù)組兩周和四周時(shí)血清E2濃度均顯著高于單純?cè)炷＝M(P0.05),四周時(shí)高劑量雌激素干預(yù)組血清TNF-a濃度顯著高于單純?cè)炷＝M(P0.05)。 2.雌激素干預(yù)組兩周和四周病灶體積顯著大于單純?cè)炷＝M(P0.05)。 3.無論是兩周還是四周,低劑量雌激素干預(yù)組甲苯胺藍(lán)染色脫顆粒／總的MC數(shù)比值顯著高于單純?cè)炷＝M(P0.05)。 4.雌激素干預(yù)組與單純?cè)炷＝M間病灶類胰蛋白酶的表達(dá)未見明顯差異(P0.05),四周時(shí)高劑量雌激素組NGF的表達(dá)顯著高于單純?cè)炷＝M(P0.05)。 5.色甘酸鈉治療組與溶劑組、空白對(duì)照組間血清E2水平無明顯差異(P0.05),色甘酸鈉高劑量治療組血清TNF-a濃度顯著低于溶劑組和空白對(duì)照組(P0.05)。 6.色甘酸鈉干預(yù)組兩周和四周時(shí)病灶體積與溶劑組、空白對(duì)照組均無明顯差異(P0.05)。 7.色甘酸鈉高劑量治療組甲苯胺藍(lán)染色活化的MC數(shù)顯著顯著低于溶劑組(P0.05),脫顆粒／總的MC數(shù)比值顯著低于溶劑組和空白對(duì)照組(P0.05)。 8.色甘酸鈉高劑量治療組類胰蛋白酶表達(dá)顯著低于溶劑組和空白對(duì)照組(P0.05),色甘酸鈉治療組NGF的表達(dá)略低于溶劑組和空白對(duì)照組,但統(tǒng)計(jì)學(xué)上無明顯差異(P0.05)。 結(jié)論 1.雌激素可促進(jìn)內(nèi)異病灶的生長,可通過激活肥大細(xì)胞,促使其脫顆粒釋放TNF-α、NGF介導(dǎo)內(nèi)異癥的發(fā)病。 2.色甘酸鈉可通過穩(wěn)定肥大細(xì)胞抑制其脫顆粒,減少TNF-α、類胰蛋白酶的釋放,緩解內(nèi)異癥癥狀。
[Abstract]:background
Endometriosis (hereinafter referred to as EMS) is a function of the growth of endometrial tissue (glands and stroma) of common gynecological diseases occur in other parts of the uterine cavity is covered outside the endometrium and uterine muscle layer, seriously affecting the health of women. The pain (including dysmenorrhea, chronic pelvic pain and dyspareunia) and infertility is the main clinical symptoms of endometriosis, and its prevalence in women of childbearing age in 10-15%. although endometriosis incidence showed increasing trend, but its etiology and pathogenesis and mechanism of endometriosis pain is still unclear. In recent years, many studies found that endometriosis lesions bleeding stimulation leads to aseptic inflammation, immune inflammation and nerve lesions lead to abnormal growth in central and peripheral nerve sensitization may be the main pathogenesis of endometriosis pain.
Mast cells (Mast cells MC) is one of the important cells in the immune inflammatory reaction, some scholars found that MC degranulation and migraine, interstitial cystitis, irritable bowel syndrome, there is a significant correlation between the symptoms of atopic dermatitis and other diseases. In neuropathic pain, peripheral nerve and nerve distribution in tissue MC can degranulation tryptase release, tumor necrosis factor (Tumor necrosis factor- TNF- alpha, alpha), neuropeptide, substance P (Substance P SP), nerve growth factor (Nerve growth, factor, NGF), cytokines and neurotransmitters such as acetylcholine activity medium causes sensitization, and sensitization of nerve the MC in turn activates the degranulation to release pain caused by substances releasing large amounts of neurotransmitters, further promote nerve sensitization, leading to persistent neuropathic pain. The study found that endometriosis lesions M The number of C increased and its activity increased, and MC in lesions was closely related to nerve fibers. The pain of endometriosis is also considered as a visceral and neuropathic pain. Therefore, MC plays a role in the pathogenesis of pain in endometriosis.
Research shows that estrogen can be combined with the MC film on the ER-a promoter calcium fast flow, promote human and mouse MC activation and degranulation to release the corresponding medium. Cromolyn sodium is MC membrane stabilizer, could inhibit the release of mediators, and its mechanism may be related to MC78-kDa protein phosphorylation of.78-kDa protein is composed of two sections and chromosome MC moesin homology, between the cell surface and intracellular regulatory framework by connecting the function in signal transduction, once the moesin structural changes in phosphorylation / dephosphorylation, will cause the position of rearrangement of the cell membrane and the structure, and may make the MC secretory granules.
Based on the above research, we designed this experiment, we establish the SD rat model of endometriosis, on this basis, we give the degranulation of MC promoter (estrogen) and inhibitor (sodium cromoglycate) intervention, observation model of lesion size change and the relation with MC degranulation, discuss MC in the pathogenesis of endometriosis the role, to provide new ideas for clinical treatment of endometriosis pain.
objective
To investigate the role of mast cells in the pathogenesis of endometriosis.
Materials and methods
60 healthy female non pregnant SD rats to establish rat abdominal wall endometriosis model by autologous transplantation of endometrium was divided into two parts: the first part (n = 8) rats after every intramuscular injection of different dose of estrogen (high dose group: Modeling and removing the ovaries after the exogenous estrogen 200 g/kg, low dose group, at the same time after ovariectomy exogenous estrogen 100 g/kg, simple model: the model without ovariectomy not exogenous estrogen), observing the rat endometriosis lesion size after two weeks, the morphology each group, 4 rats were killed at the remaining rats injected corresponding dose of estrogen to two weeks, were compared and total number of lesions in MC degranulation, tryptase, NGF expression, serum TNF-, estradiol (Estradiol, E2) level; the second part ( Respectively 10, 10, 8, 8) were observed in rats of endometriosis lesion size in rats after two weeks, followed by intraperitoneal injection of different doses of color acid sodium (20mg/kg of high dose group, low dose group 10mg/kg, solvent group, blank control group), two weeks were sacrificed the detection index, ibid.
statistical method
SPSS20.0 software was used to analyze the results. The results were expressed by X + S. Normality and homogeneity of variance were tested. The data of multiple groups were compared by one-way ANOVA. The data between two groups were compared with t test, and the test level was 0.05..
Result
1. the serum E2 concentration in the estrogen intervention group was significantly higher than that in the simple model group (P0.05) at two weeks and 4 weeks, and the serum TNF-a concentration in the high dose estrogen intervention group was significantly higher than that in the simple model group (P0.05).
2. the volume of the lesion in the two week and four weeks of the estrogen intervention group was significantly greater than that of the simple model group (P0.05).
3. either two weeks or around, low dose estrogen intervention group with toluidine blue staining degranulation / total number of MC was significantly higher than those in the model group (P0.05).
4. there was no significant difference in the expression of tryptase between the estrogen intervention group and the simple model group (P0.05), and the expression of NGF in the high dose estrogen group was significantly higher than that in the simple model group (P0.05).
There was no significant difference in serum E2 level between the 5. color sodium glycyrrhizinate treatment group and the solvent group and blank control group (P0.05). The serum TNF-a concentration in the high-dose sodium glycyrrhizinate group was significantly lower than that in the solvent group and the blank control group (P0.05).
There was no significant difference between the focus volume and the solvent group at two weeks and four weeks in the 6. color sodium glycolate intervention group, and there was no significant difference between the control group (P0.05).
7. color display MC cromoglycate high dose treatment group was significantly lower than that with toluidine blue staining activated solvent group (P0.05), MC ratio was significantly lower than the total degranulation / solvent group and blank control group (P0.05).
Tryptase expression in high dose 8. sodium glycolate group was significantly lower than that in the solvent group and blank control group (P0.05). The expression of NGF in the sodium glycyrrhizinate treatment group was slightly lower than that in the solvent group and blank control group, but there was no significant difference between them (P0.05).
conclusion
1. estrogen can promote the growth of endometriosis, and can induce the release of TNF- a by activating mast cells, and NGF mediates the pathogenesis of endometriosis.
2. sodium glycine can inhibit the degranulation by stabilizing the mast cells, reduce the release of TNF- - A, trypsin like, and relieve the symptoms of endometriosis.

【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R711.71

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 盧韻碧,江波,周漢良;沙丁胺醇與色甘酸鈉對(duì)大鼠腹腔肥大細(xì)胞脫顆粒過程中磷脂酶D活性的影響[J];中國藥理學(xué)與毒理學(xué)雜志;2001年05期

，

本文編號(hào)：1567273