本文選題：Klotho + 破骨細(xì)胞��； 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:探討轉(zhuǎn)染Klotho基因(KL)的表達(dá)對(duì)可溶性核因子κB配體的受體活化因子(sRANKL)誘導(dǎo)的RAW264.7單核巨噬細(xì)胞向破骨細(xì)胞分化的影響。方法:實(shí)驗(yàn)設(shè)計(jì)分為高表達(dá)KL組(Ad-KL組)、空載體組(AdGFP組)與空白對(duì)照組,倒置熒光顯微鏡觀察重組腺病毒轉(zhuǎn)染RAW264.7情況;實(shí)時(shí)熒光定量PCR(RT-PCR)和蛋白質(zhì)印跡法(Western blot)檢測(cè)KL在各組的表達(dá)情況;抗酒石酸酸性磷酸酶(TRAP)染色檢測(cè)KL的表達(dá)對(duì)sRANKL誘導(dǎo)RAW264.7向破骨細(xì)胞分化的形態(tài)學(xué)影響;RT-PCR和蛋白質(zhì)印跡法檢測(cè)各組破骨細(xì)胞成熟標(biāo)志物TRAP、組織蛋白酶K(CTSK)的相對(duì)表達(dá)情況。結(jié)果:Ad-KL成功轉(zhuǎn)染RAW264.7細(xì)胞,與Ad-GFP組和空白對(duì)照組比較,Ad-KL組細(xì)胞KL mRNA表達(dá)水平顯著升高且蛋白相對(duì)表達(dá)水平顯著增加(P0.01);TRAP染色顯示,Ad-KL組TRAP陽(yáng)性細(xì)胞數(shù)量顯少于Ad-GFP組和空白對(duì)照組,體積也更小;與Ad-GFP組和空白對(duì)照組比較,Ad-KL組TRAP和CTSK mRNA表達(dá)降低且蛋白相對(duì)表達(dá)水平也明顯降低(P0.01)。結(jié)論:KL的表達(dá)可以抑制s RANKL誘導(dǎo)RAW264.7向破骨細(xì)胞的分化過(guò)程。
[Abstract]:Aim: to investigate the effect of Klotho gene transfection on the differentiation of RAW264.7 mononuclear macrophages into osteoclasts induced by soluble nuclear factor 魏 B ligand receptor activating factor (sRANKL). Methods: the experimental design was divided into two groups: high expression KL group (Ad-KL group), empty vector group (AdGFP group) and blank control group (control group). The transfection of recombinant adenovirus into RAW264.7 was observed by inverted fluorescence microscope. The expression of KL in each group was detected by real-time quantitative PCR RT-PCR and Western blot. The expression of KL in osteoclasts induced by sRANKL was detected by tartrate-resistant acid phosphatase assay. RT-PCR and Western blotting were used to detect the relative expression of osteoclast maturation marker TRAPK and cathepsin CTSKK in osteoclasts. Results compared with Ad-GFP group and blank control group, the expression of KL mRNA in Ad-KL group was significantly higher than that in Ad-KL group, and the relative protein expression level was significantly increased in Ad-KL group. The results showed that the number of TRAP positive cells in Ad-KL group was significantly lower than that in Ad-GFP group and blank control group, and that in Ad-KL group was significantly higher than that in Ad-GFP group and blank control group. Compared with Ad-GFP group and blank control group, the expression of TRAP and CTSK mRNA in Ad-KL group was decreased and the relative expression level of protein was also significantly decreased in Ad-KL group. Conclusion the expression of KL can inhibit the differentiation of RAW264.7 into osteoclasts induced by s RANKL.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R580

【參考文獻(xiàn)】

相關(guān)期刊論文 前6條

1 蔣U喅，

本文編號(hào)：1977088