振动对体外培养IgA肾病人腭扁桃体细胞产生BAFF及低糖基化IgA1的影响

发布时间：2018-03-31 00:12

  本文选题：振动刺激　切入点：IgA肾病　出处：《中南大学》2014年硕士论文

【摘要】：目的：研究振动对体外培养IgA肾病和慢性扁桃体炎患者腭扁桃体单个核细胞产生BAFF、IgAl及其异常糖基化的影响。 方法：收集14例IgA肾病组(IgA nephropathy, IgAN组)患者和12例慢性扁桃体炎组(Chronic tonsillitis,CT组)患者腭扁桃体组织,其中慢性扁桃体炎组包括睡眠呼吸暂停综合征患者,本组均无尿检异常及肾脏病变。分离人腭扁桃体单个核细胞,培养24小时以稳定细胞恢复细胞活性状态,然后分为6组进行实验。A组直接分离培养上清及留取细胞作为基础值,B组不给予振动刺激作为对照组,C组给予1分钟振动刺激,D组给予3分钟振动刺激,E组给予5分钟振动刺激,F组给予10分钟振动刺激,B-F组继续培养72小时后收集培养上清及细胞。用ELISA法检测培养上清中IgAl蛋白的含量及异常糖基化IgAl的水平,Real time-PCR检测细胞B淋巴细胞刺激因子(B-cell-activation factor, BAFF) BAFF、βl,3-半乳糖基转移酶(core β1,3-galactosyltransferase, C1GALT1)、分子伴侣(core β1,3GalT-specific molecular chaperone, Cosmc)的基因表达,分别比较振动对IgAN组和CT组的影响以及不同振动时间的差异。 结果： 1.未给予振动刺激时,IgAN组患者腭扁桃体单个核细胞IgAl含量、异常糖基化IgAl的水平和BAFF mRNA的表达水平均高于CT组,差异有统计学意义(p0.001)；IgAN组患者腭扁桃体单个核细胞ClGALT1和Cosmc mRNA的表达水平均低于CT组,差异有统计学意义(p0.001)。 2.IgAN组患者腭扁桃体单个核细胞经振动刺激后,IgA1含量、异常糖基化IgA1的水平和BAFF mRNA的表达水平均高于未刺激组,差异有统计学意义(p0.05),C1GALT1和Cosmc mRNA的表达水平低于未刺激组,差异有统计学意义(p值均0.05)。CT组患者腭扁桃体单个核细胞经振动刺激后,BAFF mRNA的表达水平升高(p0.05),但IgA1含量、异常糖基化IgA1的水平及C1GALT1和Cosmc mRNA的表达水平无明显差异(p0.05)。 3.给予振动刺激1分钟后,IgAN组和CT组腭扁桃体单个核细胞BAFF基因的表达均有一定程度的升高,且升高程度无差异,这种升高可引起IgAN组IgA1分泌的增加,而CT组IgA1分泌无变化。随着振动时间的延长,CT组BAFF的表达不再升高,且IgA1含量无变化,而IgAN组BAFF的表达再次明显升高,且升高程度大于CT组,IgA1的分泌也显著增加。但IgAN组腭扁桃体单个核细胞C1GALT1和Cosmc mRNA的表达在不同时间振动刺激各组间无明显差异。 4. Pearson相关性分析显示：IgAN组腭扁桃体单个核细胞C1GALT1和Cosmc mRNA表达量与IgA1异常糖基化水平均呈负相关关系(r=-0.5867, p=0.001; r=-0.5031, p=0.003), BAFF mRNA的表达量与IgA1含量呈正相关关系(r=0.5037,p=0.012)。 结论：给予体外培养的人腭扁桃体单个核细胞振动刺激,可导致IgAN组和CT组BAFF基因的表达升高,振动超过1分钟后IgAN组BAFF升高的程度大于CT组。IgAN组振动可引起IgAl分泌增加,并通过抑制C1GALT1和Cosmc的基因表达来干扰IgAl的糖基化过程,使IgAl异常糖基化水平升高,即IgAl低糖基化。而CT组振动仅引起BAFF基因的表达升高,IgAl分泌无变化。给予体外培养的人腭扁桃体单个核细胞振动刺激,在一定程度上模拟了说话时腭扁桃体在咽喉部共振腔中受到声带产生的振动刺激。本实验表明,振动可对腭扁桃体单个核细胞产生BAFF及低糖基化IgAl造成一定影响,从而参与IgA肾病的发病及进展。图8幅,表9个,参考文献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.
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