论文部分内容阅读
目的探讨外源核苷酸对小鼠胸腺细胞凋亡的影响。方法首先建立了体内细胞凋亡模型,25只4周龄雄性小鼠腹腔注射地塞米松(25mg/kg)4、8、16、24h后取胸腺淋巴细胞进行细胞凋亡检测。确定建立体内细胞凋亡模型的条件为注射地塞米松(25mg/kg)后16h取小鼠胸腺细胞。外源核苷酸对小鼠胸腺淋巴细胞凋亡的影响,60只小鼠分成阴性对照、阳性对照、核苷酸1组、核苷酸2组,共4组,每组15只小鼠,均饲喂半纯合饲料。对照组每天灌胃生理盐水,核苷酸1组和2组分别灌胃125mg和25mg的核苷酸,连续饲养14d后阳性对照组和核苷酸组小鼠腹腔注射地塞米松溶液,诱导小鼠胸腺淋巴细胞凋亡,阴性对照组注射等量的生理盐水,16h后分离脾脏和胸腺,测定脾脏及胸腺指数,并采用琼脂糖凝胶电泳和流式分析方法检测胸腺淋巴细胞凋亡情况以及细胞游离Ca2+浓度([Ca2+]i)。结果核苷酸1组和2组小鼠的增重分别为371、401g,阴性对照组小鼠增重为274g,注射地塞米松后阳性对照组和核苷酸组小鼠的脾脏、胸腺重量以及脏器指数低于阴性对照,添加核苷酸对这些脏器指数没有明显影响;注射地塞米松后小鼠胸腺细胞DNA出现梯形条带,注射地塞米松后阳性对照组和核苷酸组小鼠胸腺细胞内Ca2+水平分别升高至16737nmol/L、19116nmol/L、18078nmol/L,高于阴性对照(10376nm
Objective To investigate the effects of exogenous nucleotides on the apoptosis of thymocytes in mice. Methods Apoptosis model was established in vivo. 25 male 4-week-old male mice were injected with dexamethasone (25mg / kg) for 4, 8, 16 and 24 hours respectively to detect thymus lymphocytes for apoptosis. Determine the establishment of the apoptosis model in vivo conditions for injection of dexamethasone (25mg / kg) after 16h mouse thymocytes. Sixty mice were divided into negative control, positive control, nucleotides 1 and 2, a total of 4 groups, 15 mice in each group, All feed semi-homozygous feed. The control group was orally administered with normal saline, and nucleotides 1 and 2 were intragastrically administered 125 mg and 25 mg nucleotides respectively. After 14 days of continuous feeding, the positive control group and nucleotide group mice were injected intraperitoneally with dexamethasone solution to induce small Rat thymus lymphocyte apoptosis, negative control group were injected with the same amount of saline, after 16h spleen and thymus were separated, the spleen and thymus index were measured, and the apoptosis of thymus lymphocytes was detected by agarose gel electrophoresis and flow cytometry Cell free Ca2 + concentration ([Ca2 +] i). Results The gain of nucleotides group 1 and group 2 were 371 and 401 g, respectively. The weight of the negative control group was 274 g. After injection of dexamethasone, the positive control group and the nucleotides group had the same weight of spleen and thymus And the organ index was lower than that of the negative control. The addition of nucleotides had no obvious effect on these organ indexes. The DNA of thymus cells appeared trapezoidal band after injection of dexamethasone, and the positive control group and nucleotide group after injection of dexamethasone The levels of Ca2 + in mouse thymocytes were increased to 16737nmol / L, 19116nmol / L and 18078nmol / L, respectively, higher than the negative control (10376nm