title: Neurotrophic Effects of Mu Bie Zi (Momordica cochinchinensis) Seed Elucidated by High-Throughput Screening of Natural Products for NGF Mimetic Effects in PC-12 Cells.
abstract: Post-mitotic central nervous system (CNS) neurons have limited capacity for regeneration, creating a challenge in the development of effective therapeutics for spinal cord injury or neurodegenerative diseases. Furthermore, therapeutic use of human neurotrophic agents such as nerve growth factor (NGF) are limited due to hampered transport across the blood brain barrier (BBB) and a large number of peripheral side effects (e.g. neuro-inflammatory pain/tissue degeneration etc.). Therefore, there is a continued need for discovery of small molecule NGF mimetics that can penetrate the BBB and initiate CNS neuronal outgrowth/regeneration. In the current study, we conduct an exploratory high-through-put (HTP) screening of 1144 predominantly natural/herb products (947 natural herbs/plants/spices, 29 polyphenolics and 168 synthetic drugs) for ability to induce neurite outgrowth in PC12 dopaminergic cells grown on rat tail collagen, over 7 days. The data indicate a remarkably rare event-low hit ratio with only 1/1144 tested substances (<111.25 µg/mL) being capable of inducing neurite outgrowth in a dose dependent manner, identified as; Mu Bie Zi, Momordica cochinchinensis seed extract (MCS). To quantify the neurotrophic effects of MCS, 36 images (n = 6) (average of 340 cells per image), were numerically assessed for neurite length, neurite count/cell and min/max neurite length in microns (µm) using Image J software. The data show neurite elongation from 0.07 ± 0.02 µm (controls) to 5.5 ± 0.62 µm (NGF 0.5 μg/mL) and 3.39 ± 0.45 µm (138 μg/mL) in MCS, where the average maximum length per group extended from 3.58 ± 0.42 µm (controls) to 41.93 ± 3.14 µm (NGF) and 40.20 ± 2.72 µm (MCS). Imaging analysis using immunocytochemistry (ICC) confirmed that NGF and MCS had similar influence on 3-D orientation/expression of 160/200 kD neurofilament, tubulin and F-actin. These latent changes were associated with early rise in phosphorylated extracellular signal-regulated kinase (ERK) p-Erk1 (T202/Y204)/p-Erk2 (T185/Y187) at 60 min with mild changes in pAKT peaking at 5 min, and no indication of pMEK involvement. These findings demonstrate a remarkable infrequency of natural products or polyphenolic constituents to exert neurotrophic effects at low concentrations, and elucidate a unique property of MCS extract to do so. Future research will be required to delineate in depth mechanism of action of MCS, constituents responsible and potential for therapeutic application in CNS degenerative disease or injury.
authors: College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Room 104, Dyson Pharmacy Building, 1520 ML King Blvd, Tallahassee, FL, 32307, USA.
authors: E
authors: E
authors: Mazzio
authors: Department of Biology, Florida A&M University, Tallahassee, FL, 32307, USA.
authors: B
authors: B
authors: Georges
authors: Department of Biology, Florida A&M University, Tallahassee, FL, 32307, USA.
authors: O
authors: O
authors: McTier
authors: College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Room 104, Dyson Pharmacy Building, 1520 ML King Blvd, Tallahassee, FL, 32307, USA. karam.soliman@famu.edu.
authors: Karam F A
authors: KF
authors: Soliman
mesh_terms: D000818:Animals; D001688:Biological Products / Q000494:pharmacology*; D002478:Cells, Cultured; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D029775:Momordica / Q000737:chemistry* / Q000378:metabolism; D020932:Nerve Growth Factor / Q000378:metabolism*; D055495:Neurogenesis / Q000187:drug effects*; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D016716:PC12 Cells; D051381:Rats
chemical_list: D001688:Biological Products; D020932:Nerve Growth Factor; D048049:Extracellular Signal-Regulated MAP Kinases
keywords: Chinese herbs; High throughput screening; NGF mimetic; NGF signaling; Nerve growth factor; Neurotrophic