吲哚美辛對(duì)大鼠創(chuàng)傷后異位骨化形成中Sclerostin及COX-2表達(dá)的影響
發(fā)布時(shí)間:2018-02-23 16:53
本文關(guān)鍵詞: 骨硬化蛋白 異位骨化 COX-2 吲哚美辛 免疫組化 出處:《遵義醫(yī)學(xué)院》2017年碩士論文 論文類(lèi)型:學(xué)位論文
【摘要】:目的:探討吲哚美辛對(duì)大鼠創(chuàng)傷后異位骨化(Heterotopic ossification,HO)形成中骨硬化蛋白(Sclerostin,SO)及COX-2的表達(dá)特點(diǎn)及其對(duì)HO形成的影響,為臨床診斷、預(yù)防和治療HO提供依據(jù)。方法:1.選取30只雄性SPF級(jí)SD(Sprague dawley)大鼠行左小腿X線(xiàn)檢查并隨機(jī)抽取5只大鼠抽血行ELISA測(cè)量血清SO水平后放回籠中,適應(yīng)性喂養(yǎng)1周后隨機(jī)分為實(shí)驗(yàn)組10只、模型組10只、對(duì)照組10只。無(wú)菌條件下,實(shí)驗(yàn)組和模型組行左后肢跟腱切斷誘導(dǎo)HO模型,對(duì)照組僅切開(kāi)皮膚未切斷跟腱。實(shí)驗(yàn)組吲哚美辛10mg/(kg·d)濃縮液灌胃4周,模型組及對(duì)照組分別給予同等容積生理鹽水灌胃。2.造模后第5、第10周,每組攝X線(xiàn)片觀(guān)察左跟腱部位密度變化及異位骨化評(píng)級(jí);每組取5只抽血行ELISA測(cè)量血清SO含量;每組處死5只取左側(cè)跟腱行大體形態(tài)觀(guān)察、HE染色及免疫組化檢測(cè)COX-2表達(dá)。數(shù)據(jù)以均數(shù)±標(biāo)準(zhǔn)差((?)±s)表示,應(yīng)用SPSS16.0統(tǒng)計(jì)軟件進(jìn)行統(tǒng)計(jì)分析,檢驗(yàn)水準(zhǔn)α=0.05。結(jié)果:1.第5周X線(xiàn)檢查結(jié)果其中模型組7只、實(shí)驗(yàn)組2只出現(xiàn)骨化影,對(duì)照組骨化影未現(xiàn)。模型組中COX-2陽(yáng)性表達(dá),與對(duì)照組比較有統(tǒng)計(jì)學(xué)意義(P0.05);而實(shí)驗(yàn)組COX-2表達(dá)弱陽(yáng)性,與模型組比較有統(tǒng)計(jì)學(xué)意義(P0.05);對(duì)照組COX-2呈現(xiàn)陰性或者弱陽(yáng)性,無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。模型組SO含量較造模前降低,且均低于對(duì)照組及實(shí)驗(yàn)組(P0.05);實(shí)驗(yàn)組SO濃度升高,與對(duì)照組比較有統(tǒng)計(jì)學(xué)意義(P0.05);對(duì)照組血清SO濃度變化無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。2.第10周X線(xiàn)各組檢查結(jié)果表明模型組5只、實(shí)驗(yàn)組2只出現(xiàn)骨化影;對(duì)照組未現(xiàn)骨化影。模型組COX-2表達(dá)強(qiáng)陽(yáng)性,與對(duì)照組比較有統(tǒng)計(jì)學(xué)意義(P0.05);實(shí)驗(yàn)組COX-2表達(dá)弱陽(yáng)性,與模型組比較有統(tǒng)計(jì)學(xué)意義(P0.05);對(duì)照組COX-2表達(dá)陰性或者弱陽(yáng)性。模型組血清SO含量較造模前降低,且低于對(duì)照組及實(shí)驗(yàn)組(P0.05);實(shí)驗(yàn)組血清SO濃度升高,與對(duì)照組比較有統(tǒng)計(jì)學(xué)意義(P0.05);對(duì)照組血清SO濃度變化無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:1.HO形成伴隨著血清SO水平下調(diào)。2.HO形成伴隨COX-2表達(dá)升高。3.吲哚美辛可抑制HO發(fā)生并可同時(shí)上調(diào)SO及下調(diào)COX-2表達(dá)。
[Abstract]:Objective: to investigate the effects of indomethacin on the expression of bone sclerosing protein (Sclerostinin so) and its effect on Ho formation in rats with heterotopic ossification (HOO) after trauma. Methods 30 male SPF grade SD(Sprague dawley rats were selected for X-ray examination of left calf, and 5 rats were randomly drawn to measure serum so level by ELISA and put back into cage. After one week of adaptive feeding, the rats were randomly divided into experimental group (n = 10), model group (n = 10) and control group (n = 10). Indomethacin (10 mg / kg 路d) concentrated solution was perfused for 4 weeks in the control group, and the model group and the control group were given the same volume of saline respectively. The rats in the model group were given the same volume of saline for 5 and 10 weeks after the establishment of the model, and the rats in the control group were given the same volume of saline for 4 weeks. The density and ectopic ossification of the left Achilles tendon were observed by X-ray film in each group, and the serum so content was measured by ELISA in 5 rats in each group. In each group, 5 left Achilles tendons were sacrificed for gross morphological observation, HE staining and immunohistochemical analysis of COX-2 expression. 鹵s): using SPSS16.0 statistical software for statistical analysis, the test level is 偽 0.05.Results: 1. The results of X-ray examination at the fifth week included 7 rats in the model group, 2 in the experimental group and 2 in the control group. The positive expression of COX-2 was found in the model group, but no ossification shadow was found in the control group. Compared with the control group, the expression of COX-2 in the experimental group was weakly positive, which was significantly higher than that in the model group, while the COX-2 in the control group was negative or weakly positive, and there was no significant difference between the control group and the control group (P 0.05). The content of so in the model group was lower than that in the model group. All of them were lower than those of the control group and the experimental group (P 0.05), the concentration of so in the experimental group was higher than that in the control group (P 0.05), and the serum so concentration in the control group had no statistical significance (P 0.05) .2.The results of X-ray examination in each group at the 10th week showed that there were 5 rats in the model group. Ossification was found in 2 rats in the experimental group, no ossification shadow in the control group. The expression of COX-2 was strongly positive in the model group, which was significantly higher than that in the control group (P 0.05), and the expression of COX-2 was weakly positive in the experimental group. Compared with the model group, the expression of COX-2 in the control group was negative or weakly positive. The serum so content in the model group was lower than that in the model group and lower than that in the control group and the experimental group, and the serum so concentration in the experimental group was higher than that in the control group. Compared with the control group, there was statistically significant difference in serum so concentration between the control group and the control group. Conclusion: 1. Ho formation is accompanied by down-regulation of serum so level. 2.Ho formation accompanied by increased COX-2 expression. Indomethacin can inhibit Ho occurrence and simultaneously. Upregulation of so and down-regulation of COX-2 expression.
【學(xué)位授予單位】:遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2017
【分類(lèi)號(hào)】:R681
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