论文部分内容阅读
目的:运用D-半乳糖建立亚急性衰老小鼠模型,在对其进行生物学鉴定的基础上,分析脾脏B细胞表面重要膜型分子的改变及其意义。方法:健康昆明鼠,颈后部皮下注射100 g/L D-半乳糖溶液,0.25 mL/20 g,1次/d,连续42 d。逐日观察动物的生存状态和生物学行为并定期称量其体质量动态变化。小鼠衰老模型的生物学鉴定采用血清中超氧化物歧化酶(SOD)活力、丙二醛(MDA)浓度的测定;脾脏细胞表面与活化、记忆相关的重要膜型分子的分析采用免疫荧光和流式细胞术;血清中细胞因子IL-4的分析采用ELISA法。结果:成功建立亚急性衰老小鼠模型。免疫荧光和流式细胞术分析模型组小鼠脾脏细胞上CD20+为56.8%,CD40+为43.6%,CD20+CD45RA+B为14.04%,CD40+CD45RA+B为35.4%,CD20+CD86+B为2.25%。CD40+CD86+B为4.38%,CD20+CD196+B为10.68%,CD40+CD196+B为10.98%。ELISA法检测结果显示模型组小鼠血清中IL-4的平均水平为7.93 ng/L。经统计学分析模型组小鼠脾脏细胞CD20、CD40、CD45RA、CD86的表达及血清中IL-4的水平均低于对照组(P<0.05),CD196高于对照组(P<0.01)。结论:在机体衰老过程中,脾脏B细胞表面与活化、记忆相关的重要膜型分子的表达发生改变,活化细胞减少,记忆细胞增多,血清中IL-4的表达下降,导致免疫系统功能紊乱。
OBJECTIVE: To establish a sub-acute aging mouse model by using D-galactose, and to analyze the changes of important membrane-type molecules on the surface of splenic B cells and its significance on the basis of biological identification. Methods: Healthy Kunming mice were injected subcutaneously with 100 g / L D-galactose solution 0.25 mL / 20 g once daily for 42 days. Observed daily animal survival and biological behavior and regularly weighed dynamic changes in body mass. The biological characteristics of the mouse model of aging were determined by measuring the activity of superoxide dismutase (SOD) and the concentration of malondialdehyde (MDA) in the serum. The analysis of important membrane-type molecules related to activation and memory of spleen cells was performed by immunofluorescence and flow cytometry Cytometry; Serum IL-4 cytokine analysis by ELISA. Results: Subacute aging mice model was successfully established. Immunofluorescence and flow cytometry analysis showed that the spleen cells in model group were 56.8% of CD20 +, 43.6% of CD40 +, 14.04% of CD20 + CD45RA + B, 35.4% of CD40 + CD45RA + B and 2.25 of CD20 + CD86 + B %. 4.38% for CD40 + CD86 + B, 10.68% for CD20 + CD196 + B and 10.98% for CD40 + CD196 + B. The results of ELISA showed that the average level of IL-4 in the serum of mice in model group was 7.93 ng / L. The expression of CD20, CD40, CD45RA and CD86 in spleen cells and the levels of IL-4 in serum in the model group were lower than those in the control group (P <0.05) and CD196 in the model group (P <0.01). CONCLUSION: During the aging process, the expression of important membrane-type molecules related to activation and memory changes in spleen B cells. The number of activated cells, memory cells and the expression of IL-4 in serum are decreased, resulting in dysfunction of the immune system.