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MPP+로 유도된 SH-SY5Y신경세포 사멸에 대한 고분자성분제거 봉독약침액의 신경보호 효과 연구

Neuroprotective Effects of Bee Venom, which Removes High Molecular Elements against MPP+-induced Human Neuroblastoma SH-SY5Y Cell Death

김승남|박지연|박히준|이혜정|권기록|배광록|두아름
대한한방내과학회 (The Society Of Korean Oriental Internal Medicine)|2010.06|대한한방내과학회지 (The Journal of Korean Oriental Internal Medicine)|제31권 제2호|pp. 254-263

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키워드

ApoptosisERKcaspase-3AKTMPP+SBVMPTPParkinson's disease(PD)SH-SY5YSweet bee venom
주제분류한의학

초록

Objectives : The neuroprotective effects of bee venom (BV) have been demonstrated in many studies, but bee venom has many side effects. So we used sweet bee venom (SBV), which has high molecular elements removed to reduce the side effects. I examined the neuroprotective effect of sweet bee venom in 1-methyl-4-phenylpyridine (MPP+)-induced human neuroblastoma SH-SY5Y cells. Methods : To observe the possible toxicity of SBV itself, SH-SY5Y cells were treated with SBV in various concentrations for 3 h and MPP+ in concentrations (1 and 5mM) for 24h. To investigate the protective effect of SBV against MPP+ toxicity, SH-SY5Y cells were pretreated with vehicle or nontoxic concentrations of SBV for 3h and the cells were not washed, followed by incubation with respective concentrations of SBV and 1 mM MPP+ for 24h. To investigate the protective effect of SBV against MPP+ toxicity, SH-SY5Y cells were pretreated with vehicle or nontoxic concentrations of SBV for 3h and the cells were not washed, followed by incubation with respective of SBV(0.5%), 1 mM MPP+, 5uM AKT inhibitor(LY984002) and 10uM ERK inhibitor(PD98059) for 24 h. The protective effect was measured by cell viability assay. To investigate the degree of apoptosis, caspase-3 enzyme activity was measured in control, MPP+, SBV+MPP+. Results : SBV (0.5%) pretreatment protected the SH-SY5Y cells against MPP+-induced apoptotic cell death. The cell viability was higher in the SH-SY5Y cells that were pretreated with vehicle or nontoxic concentrations of SBV than those not pretreated. The caspase-3 activity was lower in the pretreated groups than these not pretreated. ERK and AKT enzymes have a role in the neuroprotective effects of the sweet bee venom. Conclusions : The results demonstrate that SBV has a protective effect on dopaminergic neurons against MPP+ toxicity. This data suggest that SBV could be a potential therapeutic tool for neurodegenerative diseases such as Parkinson's disease(PD).

Abstract

Objectives : The neuroprotective effects of bee venom (BV) have been demonstrated in many studies, but bee venom has many side effects. So we used sweet bee venom (SBV), which has high molecular elements removed to reduce the side effects. I examined the neuroprotective effect of sweet bee venom in 1-methyl-4-phenylpyridine (MPP+)-induced human neuroblastoma SH-SY5Y cells. Methods : To observe the possible toxicity of SBV itself, SH-SY5Y cells were treated with SBV in various concentrations for 3 h and MPP+ in concentrations (1 and 5mM) for 24h. To investigate the protective effect of SBV against MPP+ toxicity, SH-SY5Y cells were pretreated with vehicle or nontoxic concentrations of SBV for 3h and the cells were not washed, followed by incubation with respective concentrations of SBV and 1 mM MPP+ for 24h. To investigate the protective effect of SBV against MPP+ toxicity, SH-SY5Y cells were pretreated with vehicle or nontoxic concentrations of SBV for 3h and the cells were not washed, followed by incubation with respective of SBV(0.5%), 1 mM MPP+, 5uM AKT inhibitor(LY984002) and 10uM ERK inhibitor(PD98059) for 24 h. The protective effect was measured by cell viability assay. To investigate the degree of apoptosis, caspase-3 enzyme activity was measured in control, MPP+, SBV+MPP+. Results : SBV (0.5%) pretreatment protected the SH-SY5Y cells against MPP+-induced apoptotic cell death. The cell viability was higher in the SH-SY5Y cells that were pretreated with vehicle or nontoxic concentrations of SBV than those not pretreated. The caspase-3 activity was lower in the pretreated groups than these not pretreated. ERK and AKT enzymes have a role in the neuroprotective effects of the sweet bee venom. Conclusions : The results demonstrate that SBV has a protective effect on dopaminergic neurons against MPP+ toxicity. This data suggest that SBV could be a potential therapeutic tool for neurodegenerative diseases such as Parkinson's disease(PD).

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