本文選題：基質(zhì)金屬蛋白酶　切入點：組織抑制劑　出處：《華中農(nóng)業(yè)大學》2015年碩士論文

【摘要】：基質(zhì)金屬蛋白酶2(MMP-2)是一種蛋白水解酶;它能夠水解細胞外基質(zhì)中的多種成分,其主要功能是組織和重塑ECM。MMP-2的自然組織抑制劑為TIMP-2,它可以結合不管是有活性的還是無活性的MMP-2,從而使MMP-2的活動中止,繼而有效的使MMP-2的膠原和明膠水解的活性降低了,最終抑制了癌細胞或子宮內(nèi)膜細胞的轉移和侵襲。試驗調(diào)查顯示,在正常情況下,在MMP-2和TIMP-2的相互作用下,它們是處于平衡協(xié)調(diào)的關系。白細胞介素-10是經(jīng)過Th2分泌的大分子,能夠降低Th1的功能,具有很強的抗炎作用,為抗炎因子。前列腺素E2(PGE2)是一種促炎細胞因子,它不僅能促進Th1細胞和Th1型T細胞的分化增加,進而促進炎癥反應;而且可以抑制免疫反應和細胞吞噬、中性粒細胞聚集與遷移。研究顯示,IL-10和PGE2與哺乳動物體內(nèi)MMP-2、TIMP-2存在密切的聯(lián)系,奶牛子宮內(nèi)膜ECM的降解和重組受到MMP-2與TIMP-2的密切調(diào)控。因此,研究MMP-2、TIMP-2對奶牛子宮內(nèi)膜炎的影響,可以調(diào)控奶牛子宮內(nèi)膜炎的發(fā)生發(fā)展及其疾病的防治提供新的理論基礎和實驗依據(jù)。本實驗選用健康荷斯坦奶牛的對照組和患有子宮內(nèi)膜炎荷斯坦奶牛的試驗組各10頭,采取血液以及子宮內(nèi)膜組織。用ELISA方法檢測血清白細胞介素-10與前列腺素E2。所取的子宮內(nèi)膜組織一部分用于提取總RNA,并在反轉錄之后采用熒光定量PCR法檢測基質(zhì)金屬蛋白酶-2與基質(zhì)金屬蛋白酶組織抑制劑-2的m RNA表達水平;另一部分是用于免疫組化法測定基質(zhì)金屬蛋白酶-2與基質(zhì)金屬蛋白酶組織抑制劑-2在蛋白水平上的分泌。運用SPSS Statistics 17.0分析實驗結果。實驗結果顯示:1.健康荷斯坦奶牛組中PGE2濃度為1.473103±80.29ng/L,IL-10含量為48.41±5.45 ng/L;而患有子宮內(nèi)膜炎的荷斯坦奶牛組中PGE2的濃度為0.933103±21.99ng/L,IL-10含量為78.45±2.55 ng/L。實驗組中PGE2濃度極顯著低于對照組(P0.01);而IL-10含量顯著高于對照組(P0.01)。2.試驗組子宮內(nèi)膜組織中基質(zhì)金屬蛋白酶-2與基質(zhì)金屬蛋白酶組織抑制劑-2m RNA水平均極顯著低于對照組(P0.01)。3.健康組子宮內(nèi)膜組織中基質(zhì)金屬蛋白酶-2與基質(zhì)金屬蛋白酶組織抑制劑-2的蛋白水平用IOD值表達,分別為(10.452±1.231)3103、(8.367±1.324)3103;而患有子宮內(nèi)膜炎組分別為(5.432±0.587)3103、(4.287±0.536)3103。因此,健康組中MMP-2與TIMP-2在蛋白水平表達都高于對照組,差異極顯著(P0.01)。實驗結果表明,子宮內(nèi)膜炎病牛中IL-10的含量升高、PGE2的含量降低,子宮內(nèi)膜中MMP-2、TIMP-2低表達,從而引起ECM供給奶牛子宮內(nèi)膜細胞的平衡穩(wěn)態(tài)得到了破壞,促進了對子宮內(nèi)膜炎的產(chǎn)生和發(fā)展。說明它們之間存在著密切的聯(lián)系,這還需要進一步的實驗予以證明。
[Abstract]:Matrix metalloproteinase 2 (MMP-2) is a proteolytic enzyme that hydrolyzes many components of extracellular matrix.Its main function is that the natural tissue inhibitor of tissue remodeling of ECM.MMP-2 is TIMP-2, which can combine both active and inactive MMP-2, thus suspending the activity of MMP-2 and effectively reducing the hydrolysis activity of collagen and gelatin in MMP-2.It eventually inhibits the metastasis and invasion of cancer cells or endometrial cells.The experimental results show that MMP-2 and TIMP-2 are in a balanced and coordinated relationship under normal conditions.Interleukin-10 (IL-10) is a macromolecule secreted by Th2, which can reduce the function of Th1. It has a strong anti-inflammatory effect and is an anti-inflammatory factor.Prostaglandin E _ 2 (PGE _ 2) is a pro-inflammatory cytokine, which can not only increase the differentiation of Th1 cells and Th1 T cells, but also inhibit the immune response, phagocytosis, neutrophil aggregation and migration.The results showed that IL-10 and PGE2 were closely related to MMP-2 and TIMP-2 in mammalian tissues. The degradation and recombination of ECM in the endometrium of dairy cows were closely regulated by MMP-2 and TIMP-2.Therefore, to study the effect of MMP-2 and TIMP-2 on endometritis in dairy cows can provide a new theoretical basis and experimental basis for regulating the occurrence and development of endometritis and the prevention and treatment of the disease in dairy cows.Blood and endometrial tissue were collected from 10 healthy Holstein cows and 10 endometritis Holstein cows respectively.Serum interleukin-10 and prostaglandin E _ 2 were detected by ELISA.Part of the endometrial tissue was used to extract total RNAs, and the m RNA expression levels of matrix metalloproteinase-2 and tissue inhibitor of matrix metalloproteinase-2 were detected by fluorescence quantitative PCR after reverse transcription.The other part is used to detect the secretion of matrix metalloproteinase-2 and tissue inhibitor of matrix metalloproteinase-2 at the protein level by immunohistochemical method.The experimental results were analyzed by SPSS Statistics 17.0.The result of the experiment shows that 1: 1.The concentration of PGE2 in the experimental group was significantly lower than that in the control group, while the content of IL-10 was significantly higher than that in the control group.The levels of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase (MMPs) -2m RNA in endometrium of the experimental group were significantly lower than those in the control group (P 0.01 路3).Therefore, the expression of MMP-2 and TIMP-2 in the healthy group was significantly higher than that in the control group (P 0.01).The results showed that the content of IL-10 in endometritis cattle increased and the content of PGE2 decreased, and the expression of MMP-2 and TIMP-2 in endometrium decreased, which resulted in the disruption of the homeostasis of ECM supply to the endometrial cells of dairy cows.Promote the production and development of endometritis.This indicates that there is a close relationship between them, which needs to be proved by further experiments.
【學位授予單位】：華中農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S858.23

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本文編號：1716782