本文選題：胎球蛋白-A　切入點(diǎn)：腎結(jié)石　出處：《廣西醫(yī)科大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：探討胎球蛋白-A(Fetuin-A)干預(yù)對(duì)結(jié)石晶體磷酸鈣（CaP）誘導(dǎo)人巨噬細(xì)胞表達(dá)及釋放晚期炎癥因子高遷移率蛋白（HMGB1）的抑制作用。 方法：分別采用100ug/ml CaP、100ug/ml CaP+1000ng/ml Fetuin-A、100ug/ml CaP+2500ng/ml Fetuin-A、100ug/ml CaP+5000ng/ml Fetuin-A刺激已成功誘導(dǎo)為巨噬細(xì)胞的U937，于刺激后24h分別提取各組培養(yǎng)細(xì)胞總RNA，，RT-PCR檢測(cè)細(xì)胞HMGB1mRNA表達(dá)水平；提取各組細(xì)胞總蛋白、胞漿蛋白、胞核蛋白及細(xì)胞培養(yǎng)上清蛋白，Western Blot檢測(cè)細(xì)胞HMGB1總蛋白、胞漿蛋白、胞核蛋白及細(xì)胞培養(yǎng)上清蛋白含量;干預(yù)4h后分別收集每組細(xì)胞培養(yǎng)液，ELISA法檢測(cè)早期炎癥因子IL-1β、IL-6、TNF-α及單核細(xì)胞趨化蛋白MCP-1含量； 結(jié)果：CaP+1000ng/ml Fetuin-A組、CaP+2500ng/ml Fetuin-A、CaP+5000ng/ml Fetuin-A組巨噬細(xì)胞HMGB1mRNA的表達(dá)均低于CaP晶體組，和CaP晶體組比較，差異有統(tǒng)計(jì)學(xué)意義（P0.05）；CaP+1000ng/mlFetuin-A組、CaP+2500ng/ml Fetuin-A、CaP+5000ng/ml Fetuin-A組細(xì)胞內(nèi)總HMGB1蛋白、胞漿HMGB1蛋白以及細(xì)胞上清HMGB1蛋白含量均少于CaP晶體組，而胞核HMGB1蛋白含量均高于CaP晶體組，和CaP晶體組比較，差異有統(tǒng)計(jì)學(xué)意義(P 0.05)；CaP+1000ng/ml Fetuin-A組、CaP+2500ng/ml Fetuin-A、CaP+5000ng/ml Fetuin-A組細(xì)胞培養(yǎng)上清內(nèi)早期炎癥因子IL-1β、IL-6、TNF-α及單核/巨噬細(xì)胞趨化蛋白MCP-1含量均少于CaP晶體組，和CaP晶體組比較，差異有統(tǒng)計(jì)學(xué)意義（P 0.05）； 結(jié)論：1. Fetuin-A可以降低CaP晶體誘導(dǎo)巨噬細(xì)胞HMGB1mRNA轉(zhuǎn)錄水平，抑制HMGB1蛋白的合成；2. Fetuin-A可以抑制CaP晶體誘導(dǎo)巨噬細(xì)胞HMGB1胞核向胞漿轉(zhuǎn)位及釋放，降低CaP晶體干預(yù)后細(xì)胞培養(yǎng)液中HMGB1的含量；3. Fetuin-A可以抑制CaP晶體誘導(dǎo)巨噬細(xì)胞釋放早期炎癥因子IL-1β、IL-6、TNF-α及單核/巨噬細(xì)胞趨化蛋白MCP-1，降低CaP晶體刺激后細(xì)胞培養(yǎng)上清中早期炎癥因子IL-1β、IL-6、TNF-α及單核/巨噬細(xì)胞趨化蛋白MCP-1含量。
[Abstract]:Aim: to investigate the inhibitory effect of fetuin-A on the expression of human macrophages and the release of advanced inflammatory factor high mobility protein (HMGB1) induced by calcium phosphate (CaP). Methods: the U937 cells were successfully induced to macrophage by 100ugr / ml CaPU 100ug / ml CaP 1000ng / ml Fetuin-Ang-1 / ml CaP 2500ng / ml Fetuin-Ang-1 ml CaP 5000ng / ml Fetuin-A. After 24 hours of stimulation, the total RNA-RNA RT-PCR was extracted from the cultured cells of each group to detect the expression of HMGB1mRNA, and the total protein, cytosolic protein, cytosolic protein of the cells were extracted. The contents of total protein, cytoplasmic protein, nuclear protein and supernatant protein in cell culture were determined by Western Blot. Four hours after intervention, the levels of IL-1 尾 -IL-6 TNF- 偽 and monocyte chemoattractant protein (MCP-1) were detected by Elisa in each group. Results the HMGB1mRNA expression of macrophages in the cap 2500ng / ml Fetuin-Ang-1 Fetuin-A group was lower than that in the CaP crystal group. Compared with the CaP crystal group, there was a significant difference in the total HMGB1 protein in the CaP2500ng-1 / ml Fetuin-A group and in the CAP2500ng-1 / ml Fetuin-Atuin-CaP5000ng-1 Fetuin-A group. The content of cytoplasmic HMGB1 protein and supernatant HMGB1 protein was lower than that of CaP crystal group, but the content of HMGB1 protein in nucleus was higher than that in CaP crystal group, which was higher than that in CaP crystal group. There was significant difference in the content of IL-1 尾 -IL-6TNF- 偽 and monocyte / macrophage chemoattractant protein (MCP-1) in the supernatant of the cell culture supernatant in the Fetuin-A group (P < 0.05). The difference was statistically significant compared with that in the CaP crystal group (P 0.05). Conclusion Fetuin-A can decrease the level of HMGB1mRNA transcription in macrophages induced by CaP crystal and inhibit the synthesis of HMGB1 protein. Fetuin-A can inhibit the translocation and release of HMGB1 nucleus into cytoplasm induced by CaP crystal. Fetuin-A could inhibit the release of early inflammatory factor IL-1 尾 -IL-6TNF- 偽 and monocyte / macrophage chemoattractant protein MCP-1 by CaP crystal, and decrease the supernatant of cell culture stimulated by CaP crystal, and decrease the content of HMGB1 in the cell culture medium after the intervention of CaP crystal. Fetuin-A can inhibit the release of the early inflammatory factor IL-1 尾 -tNF- 偽 and monocyte / macrophage chemotactic protein MCP-1 by CaP crystal. The content of IL-1 尾 -IL-6 TNF- 偽 and monocyte / macrophage chemoattractant protein (MCP-1).
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R692.4

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