【作者】 唐双阳；

【导师】 万艳平；

【作者基本信息】 南华大学 ， 免疫学， 2008， 硕士

【摘要】 目的:通过构建慢性疲劳综合征(CFS)小鼠模型,研究运动量对CFS小鼠免疫功能的影响,为临床上CFS运动处方的研究提供免疫学实验依据。方法:取40只雄性昆明小鼠,随机分为4组,每组10只。Ⅰ组为正常对照组,Ⅱ组为CFS对照组,Ⅲ组为CFS运动1组,Ⅳ组为CFS运动2组。让Ⅱ组、Ⅲ组与Ⅳ组小鼠在每天同一时间进行6min的力竭性游泳,持续3周,观察小鼠一般情况,构建CFS小鼠模型。然后,分别让Ⅲ组与Ⅳ组小鼠每天进行1次或2次恢复运动,持续2周后,比较体重、力竭游泳时间以及悬挂静止时间。小鼠摘眼球放血,分离血清, -70℃保存备用。颈椎脱臼处死小鼠,无菌操作取脾脏称重,计算脾脏指数;利用噻唑蓝(MTT)法检测小鼠脾NK细胞活性和淋巴细胞刺激指数(SI);通过形态学检测法计算淋巴细胞转化率;利用微量聚乙二醇(PEG)半定量法检测血清循环免疫复合物(CIC); ELISA检测脾细胞经诱生后分泌的IL-2和IFN-γ量;RT-PCR检测脾细胞中TNF-α水平。利用统计软件SPSS13.0对数据进行分析处理,P<0.05代表差异有统计学意义。结果:(1)小鼠体重分别比较Ⅰ组、Ⅱ组、Ⅲ组、Ⅳ组的造模前、造模后、治疗后体重,差异无显著性(P>0.05)。(2)小鼠行为学指标计算治疗后各组力竭时间与悬挂静止时间,Ⅱ组、Ⅲ组、Ⅳ组的力竭时间与悬挂静止时间明显长于Ⅰ组(P<0.05);Ⅲ组、Ⅳ组力竭时间短于Ⅱ组(P<0.05),且Ⅳ组悬挂静止时间短于Ⅱ组(P<0.05)。(3)小鼠脾脏指数Ⅱ组、Ⅲ组、Ⅳ组脾脏指数明显高于Ⅰ组(P<0.05);Ⅲ组、Ⅳ组脾脏指数明显低于Ⅱ组(P<0.05)。(4)小鼠血清CIC值Ⅱ组和Ⅲ组小鼠血清CIC值明显高于Ⅰ组(P<0.05);Ⅲ组与Ⅱ组比较差异无显著性(P>0.05);Ⅳ组血清CIC值明显低于Ⅱ组(P<0.05)。(5)小鼠脾脏NK细胞活性Ⅱ组和Ⅲ组小鼠NK细胞活性明显低于Ⅰ组(P<0.05);Ⅲ组小鼠虽高于Ⅱ组,但与Ⅱ组比较差异无显著性(P>0.05);Ⅳ组NK细胞活性明显高于Ⅱ组(P<0.05)。(6)小鼠淋巴细胞增殖Ⅱ组小鼠淋巴细胞的SI明显低于Ⅰ组(P<0.05);Ⅲ组、Ⅳ组与Ⅱ组比较,有显著性差异(P<0.05)。Ⅱ组和Ⅲ组淋巴细胞转化率明显低于Ⅰ组(P<0.05);Ⅲ组和Ⅳ组的淋巴细胞转化率与Ⅱ组比较,Ⅳ组明显高于Ⅱ组(P<0.05),但Ⅲ组与Ⅱ组比较差异无显著性(P>0.05)。(7)小鼠脾细胞IL-2与IFN-γ水平Ⅱ组小鼠IL- 2和IFN-γ含量明显低于Ⅰ组(P<0.05);Ⅲ组IL- 2含量与Ⅱ组比较差异无显著性(P>0.05),Ⅲ组IFN-γ含量明显高于Ⅱ组(P<0.05);Ⅳ组IL- 2和IFN-γ含量明显高于Ⅱ组(P<0.05)。(8)小鼠TNF-α检测Ⅱ组、Ⅲ组、Ⅳ组的TNF-α与Ⅰ组比较,Ⅱ组、Ⅲ组条带亮度明显比Ⅰ组亮;Ⅲ组、Ⅳ组与Ⅱ组比较,Ⅳ组条带亮度明显比Ⅱ组暗,Ⅲ组与Ⅱ组条带亮度差不多。结论:(1)成功地建立了CFS小鼠模型。(2)适度的运动能改善CFS小鼠的行为学指标与脾脏指数,降低小鼠血清CIC量,增加小鼠脾脏NK细胞活性、淋巴细胞SI、转化率,增加小鼠脾细胞IL-2、IFN-γ含量,降低TNF含量。(3)适量且有规律运动有助于CFS小鼠恢复健康。(4)本实验结果可为临床上研究CFS的运动处方作参考。更多还原

【Abstract】 Objective: To provide the experimental basis for studying the clinical exercise prescription of patients with chronic fatigue syndrome(CFS),the murine models built were used to investigated the effects of amount of exercise on the immune functions of murine models with CFS.Methods: Forty KM male mice were randomly divided four groups. They were comparative normal group(GroupⅠ, 10 mice), comparative CFS group(GroupⅡ,10 mice), exercise 1 group(GroupⅢ,10 mice) or exercise 2 group(GroupⅣ,10 mice). GroupⅡ,GroupⅢor GroupⅣwere forced to have 6-minute exhaustive swimming at the same time every day for three weeks. Then, GroupⅢor GroupⅣhad once or twice exercise for recover every day for two weeks. Weight, exhausted time or immovable time in suspension of every group were compared. After the murine eyeballs were picked, sera were abstracted and reserved in the refrigerator of -70℃. After execution by cervical vertebra dislocation, murine spleens taken with sterile operation were weighted. The spleen index were compared. The NK cells activities and lymphocytes stimulation index(SI) were measured by methyl thiazolyl tetrazolium (MTT) assay .The lymphocyte transformation rates were calculated by morphology detection. The circulating immune complexes (CIC)in sera were detected with PEG semiquantitative method. The content of inducible splenic IL-2 or IFN-γwas determined by ELISA, respectively. The splenic TNF-αwas detected by RT-PCR. The data were analyzed by statistics software SPSS13.0. When P<0.05, the difference was statistical significance.Results:(1) Murine weight Compared the weight of each group before the building of model, after the model built or post-treatment , the differences were non-significance (P>0.05).(2) Murine ethology index Calculated the immovable time in suspension or exhausted time of each group, the exhausted time or immovable time in suspension of GroupⅡ, GroupⅢor GroupⅣwere obviously longer than that of GroupⅠ( P<0.05). The exhausted time of GroupⅢor GroupⅣwas obviously shorter than that of GroupⅡ( P<0.05). The immovable time in suspension of GroupⅣwas obviously shorter than that of GroupⅡ( P<0.05).(3) Murine spleen index The spleen indexs of GroupⅡ, GroupⅢor GroupⅣwere obviously higher than that of GroupⅠ(P<0.05). The spleen indexs of GroupⅢor GroupⅣwas obviously lower than that of GroupⅡ( P<0.05).(4)Murine CIC in sera The contents of CIC in sera of GroupⅡor GroupⅢwas obviously higher than that of GroupⅠ(P<0.05). Compared GroupⅢwith GroupⅡ, the differences were non-significance(P>0.05). The contents of CIC in sera of GroupⅣwere obviously fewer than those of GroupⅡ(P<0.05).(5)The NK cells activities of mice The NK cells activities of GroupⅡor GroupⅢwere obviously lower than those of GroupⅠ(P<0.05). The NK cell activities of GroupⅢraised certainly, but were non-significance differences by compared with GroupⅡ(P>0.05). The NK cell activities of GroupⅣwere obviously higher than those of GroupⅡ(P<0.05).(6) Lymphocyte proliferation of mice The SI of murine lymphocyte of GroupⅡwas obviously lower than that of GroupⅠ(P<0.05). Compared the lymphocyte SI of GroupⅢor GroupⅣwith GroupⅡ, the differences were statistical significance(P<0.05). The lymphocyte transformation rates of GroupⅡor GroupⅢwere obviously lower than those of GroupⅠ(P<0.05). Compared the lymphocyte transformation rates of GroupⅢor GroupⅣwith GroupⅡ, those of GroupⅣwere obviously higher than those of GroupⅡ(P<0.05), but the difference were non-significance between GroupⅡand GroupⅢ(P>0.05).(7)Expression level of murine splenic IL-2 and IFN-γThe contents of splenic IL-2 or IFN-γof GroupⅡwere obviously lower than those of GroupⅠ(P<0.05). The differences were non-significance by compared the contents of IL-2 of GroupⅢwith GroupⅡ(P>0.05). The contents of IFN-γof GroupⅢwas obviously higher than that of GroupⅡ(P<0.05). The contents of murine splenic IL-2 or IFN-γof Group Ⅳwere obviously higher than those of GroupⅡ(P<0.05).(8) TNF-αdetection of mice Compared the contents of TNF-αof GroupⅡ, GroupⅢor GroupⅣwith GroupⅠ, the strap brightness of GroupⅡor GroupⅢwere obviously brighter than that of GroupⅠ. Compared GroupⅢor GroupⅣwith GroupⅡ, the strap brightness of GroupⅣwas obviously more dimmed than that of GroupⅡ. The strap brightness of GroupⅢwas nearly the same as that of GroupⅡ.Conclusions:(1) Murine models with CFS were successfully built.(2) Appropriate exercise might improve murine ethology index and spleen index, lower the contents murine CIC in sera, and increase the NK cells activities , lymphocyte SI or transformation rates. Meanwhile, it might adjust murine splenic IL-2 , IFN-γ, TNF-α.(3) Appropriate and regular exercise could be benefit for the recovery of mice with CFS.(3) The experimental results might provide the experimental basis for studying the clinical exercise prescription of the patients with CFS.更多还原