Jia Yu-Jie, Nie Kun, Zhang Xue-Zhu, Li Jing, Guo Lin
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2016 Dec;36(12):1474-1479.
Objective To observe the correlations between heat shock protein 84 ( Hsp84)/ Hsp86 and brain aging in senescence accelerated mouse prone 8 (SAMP8) and the regulation effects of acupuncture. Methods Ten senescence accelerated mouse resistant 1 (SAMR1) were recruited as a normal control group. Another 30 SAMP8 mice were divided into the blank control group, the acupuncture group, and the non-acupoint group by random digit table, 10 in each group. Mice in the acupuncture group received treatment with "Sanjiao" acupuncture method. Mice in the non-acupoint group were needled at two fixed non-acupoints located at bilateral hypochondrium of the body. Catching stimulus at equal volume was given to mice in the rest two groups. All intervention was performed once per day for 15 successive days. Neuromuscular coordination of mice was evaluated. Levels of oxidative stress and protein carbonyl were determined. mRNA and protein expression levels of Hsp84 and Hsp86 in the hippocampus of mice and Neuro-2a cells were detected using Real-time PCR and Western blot. Results Compared with the normal control group, the success rate of tight rope experiment was lowered (P <0. 01) , levels of super- oxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the hippocampus were reduced (P < 0. 01) , the levels of superoxide anion and protein carbonyl increased (P <0. 01, P <0. 05), mRNA and protein expression levels of Hsp84 and Hsp86 in the hippocampus decreased (P <0. 01) in the blank con- trol group. Compared with the blank control group and the non-acupoint group, the success rate of tight rope experiment was elevated (P <0. 05) , levels of SOD and GSH-Px were increased (P <0. 05, P < 0. 01) , the levels of superoxide anion and protein carbonyl decreased (P <0. 01 , P <0. 05), and expres- sion levels of Hsp84 and Hsp86 in the hippocampus increased (P <0. 01) in the acupuncture group. mR- NA expression levels of Hsp84 and Hsp86 were decreased in Neuro-2a cells after treated with AP21-35 (P <0. 01). Conclusions Increased oxidative damage of protein and decreased expression levels of Hsp84 and Hsp86 might be partial reasons for resulting in accumulation of denatured protein and brain aging in SAMP8. Acupuncture could delay brain aging by regulating the expressions of Hsp84 and Hsp86.
目的 观察热休克蛋白84（Hsp84）/热休克蛋白86（Hsp86）与快速老化小鼠8（SAMP8）脑老化的相关性及针刺的调节作用。方法 选取10只快速老化抗性小鼠1（SAMR1）作为正常对照组。另将30只SAMP8小鼠通过随机数字表法分为空白对照组、针刺组和非穴位组，每组10只。针刺组小鼠采用“三焦”针法治疗。非穴位组小鼠在双侧胁肋部两个固定非穴位处针刺。其余两组小鼠给予等量抓握刺激。所有干预均每天进行1次，连续进行15天。评估小鼠的神经肌肉协调性。测定氧化应激水平和蛋白质羰基含量。采用实时荧光定量聚合酶链反应（Real-time PCR）和蛋白质免疫印迹法（Western blot）检测小鼠海马及Neuro-2a细胞中Hsp84和Hsp86的mRNA和蛋白表达水平。结果 与正常对照组相比，空白对照组小鼠走钢丝实验成功率降低（P<0.01），海马中超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）水平降低（P<0.01），超氧阴离子和蛋白质羰基水平升高（P<0.01，P<0.05），海马中Hsp84和Hsp86的mRNA和蛋白表达水平降低（P<0.01）。与空白对照组和非穴位组相比，针刺组小鼠走钢丝实验成功率升高（P<0.05），SOD和GSH-Px水平升高（P<0.05，P<0.01），超氧阴离子和蛋白质羰基水平降低（P<0.01，P<0.05），海马中Hsp84和Hsp86的表达水平升高（P<0.01）。用AP21-35处理后，Neuro-2a细胞中Hsp84和Hsp86的mRNA表达水平降低（P<双0.01）。结论 蛋白质氧化损伤增加及Hsp84和Hsp86表达水平降低可能是导致SAMP8小鼠变性蛋白蓄积和脑老化的部分原因。针刺可通过调节Hsp84和Hsp86的表达延缓脑老化。
