本文選題：振動刺激　切入點(diǎn)：IgA腎病　出處：《中南大學(xué)》2014年碩士論文

【摘要】：目的：研究振動對體外培養(yǎng)IgA腎病和慢性扁桃體炎患者腭扁桃體單個核細(xì)胞產(chǎn)生BAFF、IgAl及其異常糖基化的影響。 方法：收集14例IgA腎病組(IgA nephropathy, IgAN組)患者和12例慢性扁桃體炎組(Chronic tonsillitis,CT組)患者腭扁桃體組織,其中慢性扁桃體炎組包括睡眠呼吸暫停綜合征患者,本組均無尿檢異常及腎臟病變。分離人腭扁桃體單個核細(xì)胞,培養(yǎng)24小時以穩(wěn)定細(xì)胞恢復(fù)細(xì)胞活性狀態(tài),然后分為6組進(jìn)行實(shí)驗(yàn)。A組直接分離培養(yǎng)上清及留取細(xì)胞作為基礎(chǔ)值,B組不給予振動刺激作為對照組,C組給予1分鐘振動刺激,D組給予3分鐘振動刺激,E組給予5分鐘振動刺激,F組給予10分鐘振動刺激,B-F組繼續(xù)培養(yǎng)72小時后收集培養(yǎng)上清及細(xì)胞。用ELISA法檢測培養(yǎng)上清中IgAl蛋白的含量及異常糖基化IgAl的水平,Real time-PCR檢測細(xì)胞B淋巴細(xì)胞刺激因子(B-cell-activation factor, BAFF) BAFF、βl,3-半乳糖基轉(zhuǎn)移酶(core β1,3-galactosyltransferase, C1GALT1)、分子伴侶(core β1,3GalT-specific molecular chaperone, Cosmc)的基因表達(dá),分別比較振動對IgAN組和CT組的影響以及不同振動時間的差異。 結(jié)果： 1.未給予振動刺激時,IgAN組患者腭扁桃體單個核細(xì)胞IgAl含量、異常糖基化IgAl的水平和BAFF mRNA的表達(dá)水平均高于CT組,差異有統(tǒng)計學(xué)意義(p0.001)；IgAN組患者腭扁桃體單個核細(xì)胞ClGALT1和Cosmc mRNA的表達(dá)水平均低于CT組,差異有統(tǒng)計學(xué)意義(p0.001)。 2.IgAN組患者腭扁桃體單個核細(xì)胞經(jīng)振動刺激后,IgA1含量、異常糖基化IgA1的水平和BAFF mRNA的表達(dá)水平均高于未刺激組,差異有統(tǒng)計學(xué)意義(p0.05),C1GALT1和Cosmc mRNA的表達(dá)水平低于未刺激組,差異有統(tǒng)計學(xué)意義(p值均0.05)。CT組患者腭扁桃體單個核細(xì)胞經(jīng)振動刺激后,BAFF mRNA的表達(dá)水平升高(p0.05),但I(xiàn)gA1含量、異常糖基化IgA1的水平及C1GALT1和Cosmc mRNA的表達(dá)水平無明顯差異(p0.05)。 3.給予振動刺激1分鐘后,IgAN組和CT組腭扁桃體單個核細(xì)胞BAFF基因的表達(dá)均有一定程度的升高,且升高程度無差異,這種升高可引起IgAN組IgA1分泌的增加,而CT組IgA1分泌無變化。隨著振動時間的延長,CT組BAFF的表達(dá)不再升高,且IgA1含量無變化,而IgAN組BAFF的表達(dá)再次明顯升高,且升高程度大于CT組,IgA1的分泌也顯著增加。但I(xiàn)gAN組腭扁桃體單個核細(xì)胞C1GALT1和Cosmc mRNA的表達(dá)在不同時間振動刺激各組間無明顯差異。 4. Pearson相關(guān)性分析顯示：IgAN組腭扁桃體單個核細(xì)胞C1GALT1和Cosmc mRNA表達(dá)量與IgA1異常糖基化水平均呈負(fù)相關(guān)關(guān)系(r=-0.5867, p=0.001; r=-0.5031, p=0.003), BAFF mRNA的表達(dá)量與IgA1含量呈正相關(guān)關(guān)系(r=0.5037,p=0.012)。 結(jié)論：給予體外培養(yǎng)的人腭扁桃體單個核細(xì)胞振動刺激,可導(dǎo)致IgAN組和CT組BAFF基因的表達(dá)升高,振動超過1分鐘后IgAN組BAFF升高的程度大于CT組。IgAN組振動可引起IgAl分泌增加,并通過抑制C1GALT1和Cosmc的基因表達(dá)來干擾IgAl的糖基化過程,使IgAl異常糖基化水平升高,即IgAl低糖基化。而CT組振動僅引起B(yǎng)AFF基因的表達(dá)升高,IgAl分泌無變化。給予體外培養(yǎng)的人腭扁桃體單個核細(xì)胞振動刺激,在一定程度上模擬了說話時腭扁桃體在咽喉部共振腔中受到聲帶產(chǎn)生的振動刺激。本實(shí)驗(yàn)表明,振動可對腭扁桃體單個核細(xì)胞產(chǎn)生BAFF及低糖基化IgAl造成一定影響,從而參與IgA腎病的發(fā)病及進(jìn)展。圖8幅,表9個,參考文獻(xiàn)45篇。
[Abstract]:Objective: To study the effects of vibration on the production of BAFF, IgAl and abnormal glycosylation of the palatine tonsil mononuclear cells in patients with IgA nephropathy and chronic tonsillitis in vitro.
Methods: 14 cases with IgA nephropathy group (IgA nephropathy, IgAN group) and 12 cases of patients with chronic tonsillitis group (Chronic tonsillitis, CT group) in patients with tonsillar tissue, including chronic tonsillitis group included patients with sleep apnea syndrome, the group had no abnormal urine and renal lesions. The separation of human tonsillar mononuclear cells. 24 hours of training in order to stabilize cells to restore the activity of cells, and then divided into 6 groups group.A directly from the culture supernatant and cells taken as basic value, B group does not give the vibration stimulation as control group, C group was given 1 minutes of vibration stimulation, D group was given 3 minutes of vibration stimulation, E group was given 5 minutes vibration stimulation, F group was given 10 minutes of vibration stimulation, B-F group after incubated for 72 hours. Cell culture supernatants were collected and detected by ELISA in cultured IgAl protein content in the supernatant and the abnormal glycosylation level of IgAl, Real time-PCR. Measured cell B lymphocyte stimulator (B-cell-activation factor, BAFF BAFF), beta L, 3- galactosyltransferase (core beta 1,3-galactosyltransferase, C1GALT1), molecular chaperones (core beta 1,3GalT-specific molecular chaperone, Cosmc) gene expression, effects were compared vibration of IgAN group and CT group and the difference of vibration time.
Result錛，

本文編號：1688280