太子参是一种可用于保健食品的传统中药,具有抗疲劳和调节免疫等作用,但神经保护和改善记忆作用报道较少。为探究太子参改善记忆和认知障碍的有效成分及作用机制,利用Aβ1-42脑室显微注射斑马鱼模型初步探究太子参醇提物对记忆的改善作用;CCK8试剂盒测定太子参水提物、醇提物、多糖、皂苷、环肽对Aβ25-35诱导皮层神经元存活率的影响;荧光定量PCR测定太子参环肽B(heterophyllin B, HB)对神经元中凋亡相关基因的表达,免疫细胞化学染色探究HB对神经元的保护作用;利用APP/PS1转基因小鼠探究HB对痴呆样行为和记忆的改善作用。太子参醇提物给药后,斑马鱼新臂和奖励臂探寻的潜伏期显著降低,在新臂和奖励臂游动时间和总路程增加。HB显著增加神经元存活率及β3-tubulin、MAP2阳性突起的表达,并改善APP/PS1转基因小鼠记忆障碍。综上,太子参的神经保护作用可能通过HB减少突起萎缩,从而改善认知功能障碍和记忆缺陷。
Pseudostellaria heterophylla is a kind of traditional medicine as well as healthy food, which exerts anti-fatigue and immune-regulative effects, however, the neuroprotective and memory ameliorative effects are rarely reported.To explore the effective components and their mechanism in AD, the highly efficient zebrafish AD model was performed to evaluate the neuroprotective effects of Pseudostellaria heterophylla ethanol extract, cell counting kit-8 (CCK8) was used to determine the effects of aqueous extracts, ethanol extracts, polysaccharides, saponins, and cyclopeptides of Pseudostellaria heterophylla on cortical neuronal survival.Fluorescent quantitative PCR was used to detect the expression of apoptosis-related genes in neurons after intervention with heterophyllin B (HB), and immunocytochemical staining was used to investigate the neuroprotective effects.Behavioral tests were conducted to validate the memory ameliorative effects in APP/PS1 transgenic mice.Compared with Aβ-cerebroventricular microinjection zebrafish model (CVMI), Pseudostellaria heterophylla ethanol extract treatment reduced the latency to the new arm and reward arm, increased the swimming time and total distance in the new arm and reward arm, and significantly increased the survival rate in Aβ-induced primary cultured neurons.HB significantly improved memory function in APP/PS1 transgenic mice, as well as enhanced neuronal survival and elongated neurites in cultured neurons.Collectively, HB was identified as one of the effective constituents of Pseudostellaria heterophylla, which exerts neuroprotective effects, thus leading to neural network reconstruction and memory improvement.
[1] WENG Q X, CAI X X, ZHANG F, et al.Fabrication of self-assembled Radix Pseudostellariae protein nanoparticles and the entrapment of curcumin[J].Food Chemistry, 2019, 274:796-802.
[2] HU D J, SHAKERIAN F, ZHAO J, et al.Chemistry, pharmacology and analysis of Pseudostellaria heterophylla:A mini-review[J].Chinese Medicine, 2019, 14:21.
[3] HU J, PANG W S, CHEN J L, et al.Hypoglycemic effect of polysaccharides with different molecular weight of Pseudostellaria heterophylla[J].BMC Complementary and Alternative Medicine, 2013, 13:267.
[4] YANG Q, CAI X X, HUANG M C, et al.Immunomodulatory effects of Pseudostellaria heterophylla peptide on spleen lymphocytes via a Ca2+/CaN/NFATc1/IFN-γ pathway[J].Food & Function, 2019, 10(6):3 466-3 476.
[5] CHEN C, LIANG H, WANG J Y, et al.Heterophyllin B an active cyclopeptide alleviates dextran sulfate sodium-induced colitis by modulating gut microbiota and repairing intestinal mucosal barrier via AMPK activation[J].Molecular Nutrition & Food Research, 2022, 66(17):e2101169.
[6] 汪宝军, 王和鸣, 王竹风.复方太子参颗粒促进周围神经损伤后再生的实验研究[J].福建中医学院学报, 2002, 12(4):28-30;44.
WANG B J, WANG H M, WANG Z F.Experimental study of composite TAIZISHEN granule on promoting regeneration of post-injury peripheral nerve[J].Journal of Fujian College of Traditional Chinese Medicine, 2002, 12(4):28-30;44.
[7] YANG Z Y, ZHANG C, LI X H, et al.Heterophyllin B, a cyclopeptide from Pseudostellaria heterophylla, enhances cognitive function via neurite outgrowth and synaptic plasticity[J].Phytotherapy Research, 2021, 35(9):5 318-5 329.
[8] DENG J H, FENG X Y, ZHOU L J, et al.Heterophyllin B, a cyclopeptide from Pseudostellaria heterophylla, improves memory via immunomodulation and neurite regeneration in i.c.v.Aβ-induced mice[J].Food Research International, 2022, 158:111576.
[9] SCHELTENS P, DE STROOPER B, KIVIPELTO M, et al.Alzheimer's disease[J].Lancet, 2021, 397(10284):1577-1590.
[10] WELLER J, BUDSON A.Current understanding of Alzheimer's disease diagnosis and treatment[J].F1000Research, 2018, 7:1161.
[11] THOE E S, FAUZI A, TANG Y Q, et al.A review on advances of treatment modalities for Alzheimer's disease[J].Life Sciences, 2021, 276:119129.
[12] KEVADIYA B D, OTTEMANN B M, THOMAS M B, et al.Neurotheranostics as personalized medicines[J].Advanced Drug Delivery Reviews, 2019, 148:252-289.
[13] SORIA LOPEZ J A, GONZÁLEZ H M, LÉGER G C.Alzheimer's disease[J].Handbook of Clinical Neurology, 2019, 167:231-255.
[14] BEKRIS L M, YU C G, BIRD T D, et al.Genetics of Alzheimer disease[J].Journal of Geriatric Psychiatry and Neurology, 2010, 23(4):213-227.
[15] ARMSTRONG R A.Risk factors for Alzheimer's disease[J].Folia Neuropathologica, 2019, 57(2):87-105.
[16] HOLCOMB L, GORDON M N, MCGOWAN E, et al.Accelerated Alzheimer-type phenotype in transgenic mice carrying both mutant amyloid precursor protein and presenilin 1 transgenes[J].Nature Medicine, 1998, 4(1):97-100.
[17] WENGENACK T M, WHELAN S, CURRAN G L, et al.Quantitative histological analysis of amyloid deposition in Alzheimer's double transgenic mouse brain[J].Neuroscience, 2000, 101(4):939-944.
[18] BHATTARAI P, THOMAS A K, COSACAK M I, et al.IL4/STAT6 signaling activates neural stem cell proliferation and neurogenesis upon amyloid-β42 aggregation in adult zebrafish brain[J].Cell Reports, 2016, 17(4):941-948.
[19] BASHIRZADE A A, ZABEGALOV K N, VOLGIN A D, et al.Modeling neurodegenerative disorders in zebrafish[J].Neuroscience and Biobehavioral Reviews, 2022, 138:104679.
[20] LU F, YANG H, LIN S D, et al.Cyclic peptide extracts derived from Pseudostellaria heterophylla ameliorates COPD via regulation of the TLR4/MyD88 pathway proteins[J].Frontiers in Pharmacology, 2020, 11:850.
[21] BHATTARAI P, THOMAS A K, COSACAK M I, et al.Modeling amyloid-β42 toxicity and neurodegeneration in adult zebrafish brain[J].Journal of Visualized Experiments, 2017(128):56014.
[22] KIZIL C, ILTZSCHE A, KASLIN J, et al.Micromanipulation of gene expression in the adult zebrafish brain using cerebroventricular microinjection of morpholino oligonucleotides[J].Journal of Visualized Experiments, 2013(75):e50415.
[23] WONG K H, RIAZ M K, XIE Y N, et al.Review of current strategies for delivering Alzheimer's disease drugs across the blood-brain barrier[J].International Journal of Molecular Sciences, 2019, 20(2):381.
[24] SRIVASTAVA S, AHMAD R, KHARE S K.Alzheimer's disease and its treatment by different approaches:A review[J].European Journal of Medicinal Chemistry, 2021, 216:113320.
