中文摘要：

目的探讨脾脏CD4^＋T细胞组蛋白乙酰化修饰与非肥胖型糖尿病（NOD）小鼠糖尿病肾病发生及进展的关系。方法SPF级雌性NOD小鼠24只，8周龄，随机分为4组，检测12、18、24、30周龄小鼠的随机血糖，酶联免疫吸附测定（ELISA）试剂盒检测尿白蛋白和尿肌酐。分离脾脏CD4^＋T细胞，H3、H4乙酰化试剂盒检测CIM^＋T细胞H3、H4总体乙酰化水平，实时定量PCR检测组蛋白乙酰化相关修饰酶的mRNA水平，Western印迹法验证实时定量PCR筛选出的阳性基因。结果与12周龄NOD小鼠相比，24和30周龄鼠血糖升高[（18．1±6．3）、（20．7±7．5）mmol／L比（7．2±3．1）mmol／L]，尿白蛋白／肌酐比值升高[（4．04±1．54）、（8．11±1．77）mg／g比（2．12±0．56）mg／g]，脾脏CD4^＋T细胞H3总体乙酰化水平降低[（0．068±0．023）、（0．043±0．017）比（0．127±0．036）]，H4总体乙酰化水平降低[（0．058±0．022）、（0．041±0．019）比（0．082±0．032）]，均P〈0．05。在组蛋白乙酰化修饰酶谱中，与12周龄NOD小鼠相比，24和30周龄鼠组蛋白乙酰转移酶P300的mRNA水平下降[（15．53±6．31）、（13．76±3．62）比（22．94±7．40）]，P300／CBP相关因子（PCAF）的mRNA水平也降低[（3．21±0．81）、（2．74±0．36）比（5．31±0．73）]，而HDAC5mRNA及蛋白表达水平升高（均P〈0．05）。24和30周龄中已发病的NOD小鼠H3（r=-0．590，P=0．043）、H4（r=-0．702，P=0．011）乙酰化水平与尿白蛋白／肌酐比值呈负相关，HDAC5表达水平与尿白蛋白／肌酐比值呈正相关（r=0．673，P=0．016）。结论随着NOD小鼠周龄及肾脏损伤的进展，其脾脏CD4^＋T细胞总体H3、H4乙酰化水平降低，组蛋白去乙酰化酶HDAC5表达水平升高。CD4^＋T细胞中HDAC5的异常升高与NOD小鼠肾脏损害相关。

英文摘要：

Objective To explore the relationship between histone aeetylation modification in spleen CD4^＋ T cells and diabetic nephropathy. Methods Non-obese diabetic （NOD） mice were randomly divided into 4 groups and random blood glucose at 12, 18, 24 and 30 weeks were detected. Urinary albumin and creatinine were detected by enzyme-linked immunosorbent assay （ELISA）. The global acetylation levels of H3 and H4 in CD4^＋T cells were measured by H3/H4 acetylation kit. The relevant mRNA expression level of histone acetylation modification enzymes was detected by real-time quantitative polymerase chain reaction （PCR） and the altered expression genes verified by Western blot. Results Compared with mice from 12 weeks, there were significant increases in blood glucose levels and urinary albumin/creatinine ratio （ACR） from 24 and 30 weeks （ glucose： （ 18. 1±6. 3 ） and （20. 7±7.5 ） vs （7.2±3.1 ） mmol/L, ACR： （4. 04±1. 54） and （8.11±1.77） vs （2. 12±0. 56）rag/g）. Conversely, the global acetylation levels of H3（0.068 ±0.023 and 0.043 ±0.017 vs 0. 127±0.036） and H4 （0.058 ±0.022 and 0.041 ±0.019 vs 0. 082 ± 0. 032） in spleen CD4＋ T cells from 24 and 30 weeks were obviously lower. The mRNA levels of such histone acetylases as P300 （ 15.53 ± 6. 31 and 13.76±3.62 vs 22. 94 ± 7.40 ） and P300/CBP- associated factor（PCAF） （3.21 ± 0. 81 and 2. 74 ± 0. 36 vs 5.31 ± 0.73 ） declined while the protein and mRNA of histone deacetylase 5 （ HDAC5 ） significantly increased from 24 and 30 weeks. All the above differences were statistically significant （ P 〈 0. 05 ）. The levels of H3 （ r = - 0. 590, P = 0. 043 ） and H4 （r = -0. 702, P = 0. 011 ）were negatively correlated with urinary ACR while HDAC5 level was positively correlated with urinary ACR from 24 and 30 weeks （r = 0. 673, P = 0. 016）. Conclusions With aging and progression of nephropathy in NOD mice, the global aeetylation levels of H3 and I-I4 decrease while the expression level of