Fujiwara Hironori, Yoshida Jun, Dibwe Dya Fita, Awale Suresh, Hoshino Haruka, Kohama Hiroshi, Arai Hiroyuki, Kudo Yukitsuka, Matsumoto Kinzo
Institute of Natural Medicine, University of Toyama, Toyama, Japan.
Department of Geriatric and Respiratory Medicine, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan.
J Tradit Complement Med. 2018 Dec 27;9(4):328-335. doi: 10.1016/j.jtcme.2018.12.001. eCollection 2019 Oct.
The aging-dependent activation of glycogen synthase kinase-3β (GSK-3β) has been suggested to be important in the onset of dementia. To discover novel therapeutic Kampo medicines for dementia, we examined the effects of orengedokuto (OGT; huáng lián jiědú tāng) and san'oshashinto (SST; sān huáng xiè xīn tāng) on memory deficits and GSK-3β activity in senescence-accelerated prone mice (SAMP8).
The object recognition test (ORT) and conditioned fear memory test (CFT) were employed to elucidate short-term working memory and long-term fear memory. The activity of GSK-3β and the phosphorylation of related molecules were measured using a kinase assay and Western blotting.
OGT and SST attenuated memory deficits in SAMP8 in ORT, but not in CFT. In experiments, cortical GSK-3β activity was significantly stronger in SAMP8 than in SAMR1. The enhanced cortical GSK-3β activity in SAMP8 was accompanied by a significant increase in the level of phosphorylated collapsin response mediator protein-2 (CRMP2), an important factor that is involved in the regulation of microtubule stability. OGT and SST attenuated not only increases in cortical GSK-3β activity, but also the levels of phosphorylated CRMP2 in SAMP8. In vitro experiments, flavonoids contained in these kampo medicines, inhibited GSK-3β activity in concentration-dependent manners. These results suggest that OGT and SST prevent aging-induced short-term working memory deficits by inhibiting aging-dependent elevations in the cortical GSK-3β activity and subsequent CRMP2 phosphorylation.
糖原合酶激酶-3β（GSK-3β）的衰老依赖性激活被认为在痴呆症的发病过程中起重要作用。为了发现治疗痴呆症的新型汉方药，我们研究了黄连解毒汤（OGT）和三黄泻心汤（SST）对衰老加速易患小鼠（SAMP8）记忆缺陷和GSK-3β活性的影响。
采用物体识别测试（ORT）和条件性恐惧记忆测试（CFT）来阐明短期工作记忆和长期恐惧记忆。使用激酶测定法和蛋白质印迹法测量GSK-3β的活性和相关分子的磷酸化水平。
OGT和SST减轻了SAMP8在ORT中的记忆缺陷，但在CFT中没有。在实验中，SAMP8皮质中的GSK-3β活性明显强于SAMR1。SAMP8中增强的皮质GSK-3β活性伴随着磷酸化的塌陷反应介导蛋白-2（CRMP2）水平的显著增加，CRMP2是参与微管稳定性调节的重要因子。OGT和SST不仅减轻了SAMP8皮质中GSK-3β活性的增加，还降低了磷酸化CRMP2的水平。在体外实验中，这些汉方药中含有的黄酮类化合物以浓度依赖性方式抑制GSK-3β活性。这些结果表明，OGT和SST通过抑制皮质GSK-3β活性的衰老依赖性升高以及随后的CRMP2磷酸化来预防衰老诱导的短期工作记忆缺陷。
