【作者】 赵广高；

【导师】 苏全生；

【作者基本信息】 北京体育大学 ， 运动人体科学， 2012， 博士

【摘要】 研究目的：通过4周递增负荷训练诱发大鼠Thl/Th2失衡,并用足三里穴位针刺手段干预部分训练大鼠,观察和分析JAK2/STAT4通路及其上游因子在该失衡发展过程中的变化规律,探讨大运动量训练导致Th1/Th2失衡的分子机制。研究方法：清洁级16周龄雄性SD大鼠随机分为安静对照组(C组)、游泳训练组(T组)、训练+捆绑组(TB组)与训练+捆绑针刺组(TBA组),每组根据取材时间不同又随机分为24h组与7d组2个小组。训练采用4周递增负荷游泳训练方法,针刺部位选用大鼠双侧足三里穴位。ELISA法检测心脏血血浆IFN-γ、IL-4、IL-12含量,Western Blotting法测定心脏血淋巴细胞pJAK2、pSTAT4、 JAK2、STAT4的蛋白表达。实验数据采用单变量方差分析法与Pearson相关性进行统计分析。研究结果：(1)T组大鼠血浆IFN-γ (P<0.01). IFN-γ/IL-4(P<0.05)与IL-12(P<0.01)水平均显著低于C组；训练后7d三指标均低于训练后24h时相,但均无显著性差异。T组大鼠血浆IL-4含量相比C组无显著性变化。C组与T组大鼠血浆IL-12与IFN-γ的含量显著相关(P<0.01)。(2)T组大鼠血淋巴细胞pJAK2(P<0.05)、pSTAT4(P<0.01)蛋白表达均显著低于C组；训练后7d两指标均低于训练后24h时相,但均无显著性差异。T组大鼠血淋巴细胞JAK2、STAT4蛋白表达相比C组无显著性变化。(3)与T组、TB组相比,TBA组各指标均未见显著性差异。研究结论：(1)4周递增负荷训练可有效抑制Th1型免疫反应,诱发Th1/Th2失衡。该失衡作用可在训练后维持一段时间,并呈加重趋势。(2)4周递增负荷训练可通过减少IL-12的分泌,抑制JAK2/STAT4信号通路中关键因子JAK2、 STAT4的磷酸化过程,降低Th1类细胞因子IFN-γ的合成。(3)足三里穴位针刺对4周递增负荷训练所诱发的Th1/Th2失衡与JAK2/STAT4信号通路抑制未见明显的干预作用。更多还原

【Abstract】 Objective:Thl/Th2imbalance in rats was induced by means of incremental load training, while two groups of rats under training were stimulated through acupuncture treatments in ZuSanLi (St-36). During the process, the changes of JAK2/STAT4pathway and upstream cytokines were observed and analysed so as to investigate the molecule mechanism of strenuous training induced Thl/Th2imbalance.Methods:Sixteen-week-old male Sprague-Dawley rats of clean grade were randomly divided into four groups:1) Control (group C),2) Training and normal recovery (group T),3)3) Training and30minutes immobilization during recovery (group TB),4) Training and30minutes immobilization plus acupuncture at ST-36during recovery (group TBA). Each group was further randomly divided into two sub-groups according to the time of sampling:1)24h post training and2)7d post training. Group T, TB and TBA performed incremental load swimming training5sessions per week for four weeks. In Group TB and TBA,30minutes immobilization was applied30minutes post each training session in week3and4. Group TBA also received acupuncture at acupoint St-36in both legs. The concentrations of IFN-γ, IL-4, IL-12in plasma of heart blood were measured using ELISA, while the expression of pJAK2, pSTAT4, JAK2and STAT4in lymphocytes were measured using Western Blotting technique. Univariate analysis of variance was performed to compare the mean values and Pearson correlation analysis was performed to measure the correlation between the concentration of IFN-γ and that of IL-12.Results:(1) The level of plasma IFN-γ (P<0.01), IFN-γ/IL-4(P<0.05) and IL-12(P<0.01) in group T were all significantly lower than that in group C. The concentrations of these three cytokins in plasma collected at7d post-training were lower than that collected at24h post-training, but all of them were no significantly different to each other. The concentration of plasma IL-4in group T was not significantly different compared with that in group C. The concentration of plasma IL-12correlated positively with that of plasma IFN-γ in group C and group T (P<0.01).(2) The expression of protein of pJAK2(P<0.05) and pSTAT4 (P<0.01) within T lymphocytes in group T were significantly lower than that in group C. The two cytokins within T lymphocytes collected at7d post-training were lower than that collected at24h post-training. The protein concentration of JAK2and STAT4within T lymphocytes in group T showed no significant difference compared with that in group C.(3) All these cytokins in group TBA had no significant differences compared with that in group T and group TB.Conclusions:(1) Th1-type immune response was effectively suppressed and Th1/Th2imbalance was induced in response to four weeks of incremental load training. Th1/Th2imbalance induced by training could continue for some time and went worse.(2) The protein concentrations of pJAK2and pSTAT4, as the key factors of JAK2/STAT4signaling pathway, were suppressed and less IFN-γ were produced through the decreased production of IL-12after four weeks of incremental load training.(3) Acupuncture at ST-36post training did not show significant effect on Th1/Th2imbalance and JAK2/STAT4signal pathway.更多还原