Qin Gaofeng, Dong Yunfang, Liu Zhenhong, Gong Zhuoyan, Gao Chenyan, Zheng Mingcui, Tian Meijing, He Yannan, Zhong Liqun, Wang Pengwen
Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine (BUCM), Haiyuncang No. 5 in Dongcheng District, Beijing, China.
Binzhou Medical University Hospital, Shandong, China.
Chin Med. 2021 Dec 2;16(1):128. doi: 10.1186/s13020-021-00540-0.
Shen-Zhi-Ling oral liquid (SZL) is an herbal formula known for its efficacy of nourishing "heart and spleen", and is used for the treatment and prevention of middle- and early-stage dementia. This study investigated the effects of SZL on amelioration of AD, and examined whether the underlying mechanisms from the perspective of neuroprotection are related to brain glucose metabolism.
Firstly, LC-MS/MS was used to analysis the SZL mainly enters the blood component. Then, the effects of SZL on cognitive and behavioral ability of APP/PS1 double transgenic mice and amyloid protein characteristic pathological changes were investigated by behavioral study and morphological observation. The effects of SZL on the ultrastructure of mitochondria, astrocytes, and micrangium related to cerebral glucose metabolism were observed using transmission electron microscopy. Then, micro-PET was also used to observe the effects of SZL on glucose uptake. Furthermore, the effects of SZL on insulin signaling pathway InR/PI3K/Akt and glucose transporters (GLUT1 and GLUT3) were observed by immunohistochemistry, Western-blot and RT-qPCR. Finally, the effects of SZL on brain glucose metabolism and key enzyme were observed. In vitro, the use of PI3K and/or GSK3β inhibitor to observe the effects of SZL drug-containing serum on GLUT1 and GLUT3.
In vivo, SZL could significantly ameliorate cognitive deficits, retarded the pathological damage, including neuronal degeneration, Aβ peptide aggregation, and ultrastructural damage of hippocampal neurons, improve the glucose uptake, transporters and glucolysis. Beyond that, SZL regulates the insulin signal transduction pathway the insulin signal transduction pathway InR/PI3K/Akt. Furthermore, 15% SZL drug-containing serum increased Aβ-induced insulin signal transduction-pathway related indicators and GLUT1 and GLUT3 expression in SH-SY5Y cells. The improvement of GLUT1 and GLUT3 in the downstream PI3K/Akt/GSK3β signaling pathway was reversed by the use of PI3K and/or GSK3β inhibitor.
In summary, our results demonstrated that improving glucose uptake, transport, and glycolysis in the brain may underlie the neuroprotective effects of SZL, and its potential molecular mechanism may be related to regulate the insulin signal transduction pathway.
参知灵口服液（SZL）是一种以“养心健脾”功效著称的中药配方，用于治疗和预防中早期痴呆症。本研究调查了参知灵口服液对改善阿尔茨海默病（AD）的作用，并从神经保护的角度探讨其潜在机制是否与脑葡萄糖代谢有关。
首先，采用液相色谱-串联质谱法（LC-MS/MS）分析参知灵口服液主要进入血液的成分。然后，通过行为学研究和形态学观察，研究参知灵口服液对APP/PS1双转基因小鼠认知和行为能力以及淀粉样蛋白特征性病理变化的影响。利用透射电子显微镜观察参知灵口服液对与脑葡萄糖代谢相关的线粒体、星形胶质细胞和微血管超微结构的影响。此外，还采用微型正电子发射断层扫描（micro-PET）观察参知灵口服液对葡萄糖摄取的影响。此外，通过免疫组织化学、蛋白质免疫印迹法（Western-blot）和实时定量聚合酶链反应（RT-qPCR）观察参知灵口服液对胰岛素信号通路InR/PI3K/Akt和葡萄糖转运蛋白（GLUT1和GLUT3）的影响。最后，观察参知灵口服液对脑葡萄糖代谢和关键酶的影响。在体外，使用磷脂酰肌醇-3激酶（PI3K）和/或糖原合成酶激酶-3β（GSK3β）抑制剂观察含参知灵口服液血清对GLUT1和GLUT3的影响。
在体内，参知灵口服液可显著改善认知缺陷，延缓病理损伤，包括神经元变性、Aβ肽聚集和海马神经元超微结构损伤，提高葡萄糖摄取、转运和糖酵解。除此之外，参知灵口服液调节胰岛素信号转导通路InR/PI3K/Akt。此外，15%含参知灵口服液血清增加了Aβ诱导的SH-SY5Y细胞中胰岛素信号转导通路相关指标以及GLUT1和GLUT3的表达。使用PI3K和/或GSK3β抑制剂可逆转下游PI3K/Akt/GSK3β信号通路中GLUT1和GLUT3的改善。
总之，我们的结果表明，改善脑内葡萄糖摄取、转运和糖酵解可能是参知灵口服液神经保护作用的基础，其潜在分子机制可能与调节胰岛素信号转导通路有关。
