温度敏感性离子通道TRPV3在人类膝关节软骨及滑膜组织中的表达

发布时间：2018-06-15 10:02

本文选题：温度敏感性离子通道TRPV3 + 软骨　；参考：《山西医科大学》2015年硕士论文

【摘要】：目的:检测温度敏感性离子通道TRPV3(thermosensitive ion channnel TRPV3)在人的膝关节滑膜及软骨标本中的分布,对人滑膜及软骨组织中TRPV3所起的作用进行研究。方法:集取各种年龄人正常的膝关节滑膜及软骨分别10例和5例,膝骨关节炎软骨及滑膜组织各5例,每例软骨组织均取自胫骨平台,滑膜组织标本均取自关节囊内层,分装在-70℃冰箱中。番红“O”染色证实软骨标本有无退行性变。首先分出5例OA软骨标本为OA组,取5例60-69岁为正常对照组,运用Western blot、RT-PCR检测正常和OA软骨、滑膜中TRPV3有无表达分布及趋势,再用免疫荧光和免疫组化方法测得TRPV3在正常及OA软骨中分布表达以及趋势。然后Western blot检测正常软骨标本,所得条带测灰度值,分30-39岁组和60-69岁组各5例,最后采取Western blot检测TRPV3位于两组软骨中的趋势。结果:番红“O”染色OA软骨标本显微镜观察异常,色度及分层模糊,细胞之间着色较低。正常软骨标本染色均匀,浅层到深层染色由浅至深,显示无退行性改变;RT-PCR、Western blot、免疫荧光与免疫组化方法在软骨及滑膜组织中都有TRPV3表达,免疫荧光检测TRPV3在软骨细胞膜上有表达,且OA较正常软骨表达较强;考马斯亮蓝染色控制上样量,检测TRPV3条带灰度值分析,结果显示TRPV3的表达量OA组高于正常对照组,t检验有统计意义(P0.05)。测得两个年龄组正常软骨标本TRPV3的表达量,结果表明TRPV3的表达随年龄的增加而升高,t检验有统计意义(p0.05)。结论:TRPV3与骨关节炎有着密切的联系,在骨关节炎的发生发展当中起一定作用,为关节软骨损伤的防治提供新靶点。
[Abstract]:Objective: to detect the distribution of temperature sensitive ion channel TRPV3 (thermosensitive ion channnel TRPV3) in human knee joint synovium and cartilage specimens, and to study the role of TRPV3 in human synovium and cartilage tissue. Methods: 10 cases of normal knee joint and 5 cases of articular cartilage of the knee and osteoarthritis cartilage of all ages were collected. And 5 cases of synovial tissue, each case of cartilage tissue was taken from the tibial plateau, and the synovial tissue specimens were taken from the inner layer of the joint capsule. It was installed in the -70 C fridge. "O" staining showed no degeneration of cartilage specimens. First, 5 cases of OA cartilage specimens were divided into group OA, 5 cases were 60-69 years old as normal control group, and Western blot, RT-PCR was used to detect normal. The expression distribution and trend of TRPV3 in OA cartilage and synovial membrane were observed, and the distribution and trend of TRPV3 in normal and OA cartilage were measured by immunofluorescence and immunohistochemistry. Then Western blot was used to detect normal cartilage specimens, and the gray value of the strip was measured, and 5 cases were divided into 30-39 years old group and 60-69 year old group. Finally, Western blot was used to detect TRPV3. The trend in the two groups of cartilage. Results: the "O" stained OA cartilage specimens were observed by microscope, the chromaticity and stratification were blurred, the coloring between cells was low. The normal cartilage specimens were dyed evenly, the superficial to deep deep staining was from shallow to deep, showing no degenerative changes; RT-PCR, Western blot, immunofluorescence and immunohistochemical methods in cartilage and synovial groups TRPV3 expression was found in all the tissues, and the expression of TRPV3 on the chondrocyte membrane was expressed by immunofluorescence, and the expression of OA was stronger than that of the normal cartilage. Coomassie brilliant blue staining was used to control the sample quantity and to detect the gray value of the TRPV3 strip. The results showed that the OA group of TRPV3 was higher than the normal control group, and the t examination was statistically significant (P0.05). The two age groups were normal soft. The expression of TRPV3 in bone specimen shows that the expression of TRPV3 increases with the increase of age and t test has statistical significance (P0.05). Conclusion: TRPV3 has a close relationship with osteoarthritis, which plays a role in the development of osteoarthritis and provides new targets for the prevention and treatment of articular cartilage injury.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R684.3

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