糖尿病的发生与免疫指标的相关性研究

The correlation between diabetes mellitus and immune index

【目的】观察在糖尿病发生过程中体液免疫与细胞免疫的相关性.【方法】通过高脂饮食来制备小鼠前期糖尿病(Pre-DM)模型,及高脂饮食联合小剂量多次注射链脲佐菌素(STZ)制备小鼠Ⅱ型糖尿病(T2DM)模型.将80只C57BL/6J试验小鼠随机分为4组:根据预试验结果,饲喂高糖高脂饮食,选取0 d(A组)、40 d(B组)、80 d的(C组),为Pre-DM组;在80 d之后用高脂饲喂联合小剂量多次注射STZ制备T2DM模型组(D组).进行空腹血糖腹腔注射糖耐量实验(IPGTT);测定相关血清生化指标,用ELISA法测定胰岛素水平及免疫指标.用比浊法作hs-CRP,应用流式细胞术检测CD_3^+、CD_3^+CD_8^+、CD_3^+CD_4^+等T淋巴细胞亚群的比例.【结果】IPGTT结果显示,D组与其他3组曲线下面积差异极显著(P<0.01).A～D组血糖存在依次上升趋势,组间均有显著差异(P<0.05),与糖尿病相关的生化指标组间均有差异性(P<0.05),CD_3^+CD_8^+与CD_3^+CD_4^+组间差异均显著(P<0.05),CD_3^+A组与D组的差异显著(P<0.05),其余各组间差异不显著(P>0.05).HE染色结果发现,T2DM的发生过程会伴随器官的损伤.【结论】Pre-DM模型组已经存在胰岛素抵抗与胰岛受损,且伴随着肝脏不同程度的损伤;Pre-DM模型组相关的免疫指标异常及T淋巴细胞亚群的比例失衡与T2DM的发生风险有一定的相关性.

【Objective】Pre-DM(pre-DM)model and high-fat diet combined with streptozotocin(STZ)at low doses were used to prepare type 2 diabetes mellitus(T2DM)models for observation the relevance of humors immunity and cellular immunity during the development of diabetes.【Method】80 C57BL/6J mice were randomly divided into 4 groups:Pre-DM model preparation,according to the results of pre-test analysis,the mouse during the modeling process,select 0 days(A group),40 days(B group),80 days(C group),Pre-DM.The T2DM model(D group)was prepared after 80 days of high-fat feeding combined with low-dose multiple injections of streptozotocin.Weekly fasting glucose glucose tolerance test(IPGTT)was performed;serum was used as a biochemical indicator,and insulin levels and immune parameters such as IL-6 and TNF-αwere determined by ELISA.The proportion of T lymphocyte subsets such as CD3^+,CD3^+CD8^+,and CD3^+CD4^+was measured by turbidimetric method for hs-CRP and flow cytometry.【Result】IPGTT results showed that the area under the curves of D group and the first three groups was significantly different(P<0.01).The blood glucose in the A^D group increased in turn and there was a difference between groups(P<0.05).Diabetes-related biochemical indicators were significantly different between groups(P<0.05),and there was a significant difference between CD3^+CD8^+and CD3^+CD4^+groups(P<0.05).There was a significant difference between CD 3+A and D groups(P<0.05).The difference between the other groups was not significant(P>0.05).HE results showed that the occurrence of T2DM was accompanied by organ damage.【Conclusion】Pre-DM already has insulin resistance and islet damage,and is accompanied by varying degrees of liver damage.The imbalance of Pre-DM-related immune parameters and the imbalance of T-lymphocyte subsets are associated with the risk of T2DM.