TNF-α和IL-6在老年性聾動(dòng)物模型衰老過(guò)程中的表達(dá)
發(fā)布時(shí)間：2018-12-13 22:06
【摘要】：目的:研究促炎性因子TNF-α和IL-6在不同年齡組老年性聾動(dòng)物模型中的表達(dá)差異及其在老年性聾發(fā)病過(guò)程中的作用及意義。 方法: 1、取18只C57BL/6J小鼠按年齡不同分為4-、20-、36-40-周齡組三組。 2、應(yīng)用逆轉(zhuǎn)錄多聚酶鏈反應(yīng)(reverse transcription-polymerase chain reaction, RT-PCR)檢測(cè)TNF-α和IL-6在4-、20-、36-40-周齡組C57BL/6J小鼠耳蝸的表達(dá)。 3、應(yīng)用免疫組織化學(xué)技術(shù)分別檢測(cè)TNF-α和IL-6在4-、20-、36-40-周齡組C57BL/6J小鼠耳蝸的表達(dá)及分布。 4、應(yīng)用組織學(xué)HE (hematoxylin and eosin staining ,HE)染色觀察不同年齡小鼠耳蝸的組織形態(tài)差異。 結(jié)果: 1、HE染色結(jié)果顯示4-周齡組和20-周齡組C57BL/6J小鼠耳蝸形態(tài)大致正常,而36-40-周齡組小鼠耳蝸重要結(jié)構(gòu)形態(tài)明顯異常,如螺旋器細(xì)胞缺失、形態(tài)異常,血管紋萎縮和螺旋神經(jīng)元數(shù)目減少。 2、TNF-α半定量RT-PCR電泳結(jié)果顯示:TNF-αmRNA在4-周齡組C57BL/6J小鼠耳蝸表達(dá)微弱,但在20-周齡組和36-40-周齡組小鼠耳蝸表達(dá)明顯增強(qiáng)。凝膠電泳圖光密度分析結(jié)果:4-周齡組:0.107±0.014;20-周齡組:0.364±0.061;36-40-周齡組:0.584±0.042,三組之間差異均具有統(tǒng)計(jì)學(xué)意義(P0.001)。 3、IL-6 RT-PCR電泳結(jié)果顯示:IL-6 mRNA在4-周齡組C57BL/6J小鼠耳蝸幾乎沒有表達(dá),但在20-周齡組和36-40-周齡組小鼠耳蝸表達(dá)明顯增強(qiáng)。凝膠電泳圖光密度分析結(jié)果:4-周齡組:0.015±0.009;20-周齡組:0.358±0.027;36-40-周齡組:0.528±0.061,三組之間差異均具有統(tǒng)計(jì)學(xué)意義(P0.001)。 4、TNF-α免疫組化結(jié)果顯示: TNF-α在4-周齡組C57BL/6J小鼠耳蝸組織表達(dá)呈弱陽(yáng)性,20-周齡組和36-40-周齡組表達(dá)呈強(qiáng)陽(yáng)性,陽(yáng)性細(xì)胞主要分布在螺旋器、血管紋和螺旋神經(jīng)元。光密度值比值分析結(jié)果:4-周齡組:0.0034±0.0009;20-周齡組:0.1362±0.0241;36-40-周齡組:0.1920±0.0328,三組之間差異均具有統(tǒng)計(jì)學(xué)意義(P0.01)。 5、IL-6免疫組化結(jié)果顯示: IL-6在4-周齡組幾乎沒有表達(dá),20-齡組和36-40-齡組表達(dá)呈強(qiáng)陽(yáng)性,陽(yáng)性細(xì)胞的分布與TNF-α的分布相一致。光密度值比值分析結(jié)果:4-周齡組:0.0008±0.0002;20-周齡組:0.1459±0.0273;36-40-周齡組:0.1868±0.0356,三組間差異均具有統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論: 1、與4-周齡組和20-周齡組C57BL/6J小鼠相比,36-40-周齡組耳蝸組織形態(tài)發(fā)生明顯的退行性改變,包括螺旋器結(jié)構(gòu)紊亂、細(xì)胞缺失,血管紋萎縮、變薄,螺旋神經(jīng)元細(xì)胞數(shù)目減少。 2、TNF-α和IL-6在老年C57BL/6J小鼠耳蝸組織中表達(dá)明顯增加,陽(yáng)性表達(dá)主要分布在Corti器、血管紋、螺旋神經(jīng)元。 3、炎性衰老可能與老年性聾的發(fā)病機(jī)制密切相關(guān),而促炎性細(xì)胞因子TNF-α和IL-6在老年性聾的發(fā)病過(guò)程中可能發(fā)揮重要作用。
[Abstract]:Aim: to study the expression of proinflammatory cytokines TNF- 偽 and IL-6 in different age groups and their roles and significance in the pathogenesis of presbycusis. Methods: 1. Eighteen C57BL/6J mice were divided into 3 groups according to their age. 2. Reverse transcriptase polymerase chain reaction (reverse transcription-polymerase chain reaction, RT-PCR) was used to detect the expression of TNF- 偽 and IL-6 in cochlea of C57BL/6J mice in 4-20 ~ (36 ~ 40) weeks group. 3. The expression and distribution of TNF- 偽 and IL-6 in the cochlea of C57BL/6J mice were detected by immunohistochemistry. 4. Histological HE (hematoxylin and eosin staining, HE) staining was used to observe the histomorphology of cochlea in mice of different ages. Results: 1the results of HE staining showed that the cochlea morphology of 4-week-old and 20-week-old C57BL/6J mice was normal, while the cochlea morphology of 36-40-week-old mice was obviously abnormal, such as the absence of spiral organ cells and the abnormal morphology of cochlea. Atrophy of stria vascularis and reduction of the number of spiral neurons. 2TNF- 偽 semi-quantitative RT-PCR electrophoresis showed that the expression of TNF- 偽 mRNA was weak in the cochlea of the 4-week-old C57BL/6J mice, but increased significantly in the 20-week-old and 36-40-week-old groups. The results of optical density analysis of gel electrophoresis: 4- week group: 0.107 鹵0.01420- week group: 0.364 鹵0.061n36-40- week group: 0.584 鹵0.042. The differences among the three groups were statistically significant (P0.001). 3The results of IL-6 RT-PCR electrophoresis showed that the expression of IL-6 mRNA in the cochlea of 4-week-old C57BL/6J mice was almost unchanged, but the expression of IL-6 mRNA was significantly increased in 20-week-old and 36-40-week-old mice cochlea. The results of optical density analysis of gel electrophoresis showed that the difference among the three groups was statistically significant (0.015 鹵0.009) and 0.358 鹵0.027 ~ 36-40 ~ (-week), 0.528 鹵0.061, respectively (P0.001). The expression of TNF- 偽 was weakly positive in the cochlea of 4-week-old C57BL/6J mice, strongly positive in 20-week-old group and 36-40-week-old group, and the positive cells were mainly distributed in the spiral organ. Stria vascularis and spiral neurons. The results of optical density ratio analysis showed that there were significant differences among the three groups (0.0034 鹵0.0009 ~ 20- week: 0.1362 鹵0.0241), 36-40 ~ (-week): 0.1920 鹵0.0328 (P0.01). The results of immunohistochemistry showed that IL-6 was almost not expressed in 4-week-old group, but strongly positive in 20-year-old group and 36-40- age group. The distribution of positive cells was consistent with that of TNF- 偽. The results of optical density ratio analysis: 4- week group: 0.0008 鹵0.0002 + 20-week-age group: 0.1459 鹵0.0273c36-40-week-age group: 0.1868 鹵0.0356. the differences among the three groups were statistically significant (P0.05). Conclusion: 1.Compared with 4-week-old and 20-week-old C57BL/6J mice, the cochlear histomorphology in 36-40-week-old group showed obvious degenerative changes, including spiral organ structure disorder, cell loss, vascular stria atrophy and thinning. The number of spiral neurons decreased. 2the expression of TNF- 偽 and IL-6 in cochlea of aged C57BL/6J mice was significantly increased, and the positive expression was mainly distributed in the Corti organ, stria vascularis and spiral neurons. 3. Inflammatory aging may be closely related to the pathogenesis of presbycusis, and proinflammatory cytokines TNF- 偽 and IL-6 may play an important role in the pathogenesis of presbycusis.
【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】：2011
【分類號(hào)】：R764.43
本文編號(hào)：2377327
[Abstract]:Aim: to study the expression of proinflammatory cytokines TNF- 偽 and IL-6 in different age groups and their roles and significance in the pathogenesis of presbycusis. Methods: 1. Eighteen C57BL/6J mice were divided into 3 groups according to their age. 2. Reverse transcriptase polymerase chain reaction (reverse transcription-polymerase chain reaction, RT-PCR) was used to detect the expression of TNF- 偽 and IL-6 in cochlea of C57BL/6J mice in 4-20 ~ (36 ~ 40) weeks group. 3. The expression and distribution of TNF- 偽 and IL-6 in the cochlea of C57BL/6J mice were detected by immunohistochemistry. 4. Histological HE (hematoxylin and eosin staining, HE) staining was used to observe the histomorphology of cochlea in mice of different ages. Results: 1the results of HE staining showed that the cochlea morphology of 4-week-old and 20-week-old C57BL/6J mice was normal, while the cochlea morphology of 36-40-week-old mice was obviously abnormal, such as the absence of spiral organ cells and the abnormal morphology of cochlea. Atrophy of stria vascularis and reduction of the number of spiral neurons. 2TNF- 偽 semi-quantitative RT-PCR electrophoresis showed that the expression of TNF- 偽 mRNA was weak in the cochlea of the 4-week-old C57BL/6J mice, but increased significantly in the 20-week-old and 36-40-week-old groups. The results of optical density analysis of gel electrophoresis: 4- week group: 0.107 鹵0.01420- week group: 0.364 鹵0.061n36-40- week group: 0.584 鹵0.042. The differences among the three groups were statistically significant (P0.001). 3The results of IL-6 RT-PCR electrophoresis showed that the expression of IL-6 mRNA in the cochlea of 4-week-old C57BL/6J mice was almost unchanged, but the expression of IL-6 mRNA was significantly increased in 20-week-old and 36-40-week-old mice cochlea. The results of optical density analysis of gel electrophoresis showed that the difference among the three groups was statistically significant (0.015 鹵0.009) and 0.358 鹵0.027 ~ 36-40 ~ (-week), 0.528 鹵0.061, respectively (P0.001). The expression of TNF- 偽 was weakly positive in the cochlea of 4-week-old C57BL/6J mice, strongly positive in 20-week-old group and 36-40-week-old group, and the positive cells were mainly distributed in the spiral organ. Stria vascularis and spiral neurons. The results of optical density ratio analysis showed that there were significant differences among the three groups (0.0034 鹵0.0009 ~ 20- week: 0.1362 鹵0.0241), 36-40 ~ (-week): 0.1920 鹵0.0328 (P0.01). The results of immunohistochemistry showed that IL-6 was almost not expressed in 4-week-old group, but strongly positive in 20-year-old group and 36-40- age group. The distribution of positive cells was consistent with that of TNF- 偽. The results of optical density ratio analysis: 4- week group: 0.0008 鹵0.0002 + 20-week-age group: 0.1459 鹵0.0273c36-40-week-age group: 0.1868 鹵0.0356. the differences among the three groups were statistically significant (P0.05). Conclusion: 1.Compared with 4-week-old and 20-week-old C57BL/6J mice, the cochlear histomorphology in 36-40-week-old group showed obvious degenerative changes, including spiral organ structure disorder, cell loss, vascular stria atrophy and thinning. The number of spiral neurons decreased. 2the expression of TNF- 偽 and IL-6 in cochlea of aged C57BL/6J mice was significantly increased, and the positive expression was mainly distributed in the Corti organ, stria vascularis and spiral neurons. 3. Inflammatory aging may be closely related to the pathogenesis of presbycusis, and proinflammatory cytokines TNF- 偽 and IL-6 may play an important role in the pathogenesis of presbycusis.
【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】：2011
【分類號(hào)】：R764.43
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