TNF-α和IL-6在老年性聋动物模型衰老过程中的表达
发布时间：2018-12-13 22:06
【摘要】：目的:研究促炎性因子TNF-α和IL-6在不同年龄组老年性聋动物模型中的表达差异及其在老年性聋发病过程中的作用及意义。 方法: 1、取18只C57BL/6J小鼠按年龄不同分为4-、20-、36-40-周龄组三组。 2、应用逆转录多聚酶链反应(reverse transcription-polymerase chain reaction, RT-PCR)检测TNF-α和IL-6在4-、20-、36-40-周龄组C57BL/6J小鼠耳蜗的表达。 3、应用免疫组织化学技术分别检测TNF-α和IL-6在4-、20-、36-40-周龄组C57BL/6J小鼠耳蜗的表达及分布。 4、应用组织学HE (hematoxylin and eosin staining ,HE)染色观察不同年龄小鼠耳蜗的组织形态差异。 结果: 1、HE染色结果显示4-周龄组和20-周龄组C57BL/6J小鼠耳蜗形态大致正常,而36-40-周龄组小鼠耳蜗重要结构形态明显异常,如螺旋器细胞缺失、形态异常,血管纹萎缩和螺旋神经元数目减少。 2、TNF-α半定量RT-PCR电泳结果显示:TNF-αmRNA在4-周龄组C57BL/6J小鼠耳蜗表达微弱,但在20-周龄组和36-40-周龄组小鼠耳蜗表达明显增强。凝胶电泳图光密度分析结果:4-周龄组:0.107±0.014;20-周龄组:0.364±0.061;36-40-周龄组:0.584±0.042,三组之间差异均具有统计学意义(P0.001)。 3、IL-6 RT-PCR电泳结果显示:IL-6 mRNA在4-周龄组C57BL/6J小鼠耳蜗几乎没有表达,但在20-周龄组和36-40-周龄组小鼠耳蜗表达明显增强。凝胶电泳图光密度分析结果:4-周龄组:0.015±0.009;20-周龄组:0.358±0.027;36-40-周龄组:0.528±0.061,三组之间差异均具有统计学意义(P0.001)。 4、TNF-α免疫组化结果显示: TNF-α在4-周龄组C57BL/6J小鼠耳蜗组织表达呈弱阳性,20-周龄组和36-40-周龄组表达呈强阳性,阳性细胞主要分布在螺旋器、血管纹和螺旋神经元。光密度值比值分析结果:4-周龄组:0.0034±0.0009;20-周龄组:0.1362±0.0241;36-40-周龄组:0.1920±0.0328,三组之间差异均具有统计学意义(P0.01)。 5、IL-6免疫组化结果显示: IL-6在4-周龄组几乎没有表达,20-龄组和36-40-龄组表达呈强阳性,阳性细胞的分布与TNF-α的分布相一致。光密度值比值分析结果:4-周龄组:0.0008±0.0002;20-周龄组:0.1459±0.0273;36-40-周龄组:0.1868±0.0356,三组间差异均具有统计学意义(P0.05)。 结论: 1、与4-周龄组和20-周龄组C57BL/6J小鼠相比,36-40-周龄组耳蜗组织形态发生明显的退行性改变,包括螺旋器结构紊乱、细胞缺失,血管纹萎缩、变薄,螺旋神经元细胞数目减少。 2、TNF-α和IL-6在老年C57BL/6J小鼠耳蜗组织中表达明显增加,阳性表达主要分布在Corti器、血管纹、螺旋神经元。 3、炎性衰老可能与老年性聋的发病机制密切相关,而促炎性细胞因子TNF-α和IL-6在老年性聋的发病过程中可能发挥重要作用。
[Abstract]:Aim: to study the expression of proinflammatory cytokines TNF- 伪 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- 伪 and IL-6 in cochlea of C57BL/6J mice in 4-20 ~ (36 ~ 40) weeks group. 3. The expression and distribution of TNF- 伪 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- 伪 semi-quantitative RT-PCR electrophoresis showed that the expression of TNF- 伪 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 卤0.01420- week group: 0.364 卤0.061n36-40- week group: 0.584 卤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 卤0.009) and 0.358 卤0.027 ~ 36-40 ~ (-week), 0.528 卤0.061, respectively (P0.001). The expression of TNF- 伪 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 卤0.0009 ~ 20- week: 0.1362 卤0.0241), 36-40 ~ (-week): 0.1920 卤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- 伪. The results of optical density ratio analysis: 4- week group: 0.0008 卤0.0002 + 20-week-age group: 0.1459 卤0.0273c36-40-week-age group: 0.1868 卤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- 伪 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- 伪 and IL-6 may play an important role in the pathogenesis of presbycusis.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R764.43
本文编号：2377327
[Abstract]:Aim: to study the expression of proinflammatory cytokines TNF- 伪 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- 伪 and IL-6 in cochlea of C57BL/6J mice in 4-20 ~ (36 ~ 40) weeks group. 3. The expression and distribution of TNF- 伪 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- 伪 semi-quantitative RT-PCR electrophoresis showed that the expression of TNF- 伪 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 卤0.01420- week group: 0.364 卤0.061n36-40- week group: 0.584 卤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 卤0.009) and 0.358 卤0.027 ~ 36-40 ~ (-week), 0.528 卤0.061, respectively (P0.001). The expression of TNF- 伪 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 卤0.0009 ~ 20- week: 0.1362 卤0.0241), 36-40 ~ (-week): 0.1920 卤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- 伪. The results of optical density ratio analysis: 4- week group: 0.0008 卤0.0002 + 20-week-age group: 0.1459 卤0.0273c36-40-week-age group: 0.1868 卤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- 伪 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- 伪 and IL-6 may play an important role in the pathogenesis of presbycusis.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R764.43
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