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정신구,오윤서,조이슬,정소영,조효문,이준식,오원근,조태오,조광원 한국통합생물학회 2017 Animal cells and systems Vol.21 No.2
The brown alga Undaria pinnatifida, which is called Mi-Yoek in Korea, has been traditionally consumed as a health food in East Asian countries. Recent studies have reported that U. pinnatifida has beneficial effects on arteriosclerosis, inflammation, fat metabolism, and tumors. In this study, we examined the anti-senescence effects of ethanol extracts of U. pinnatifida (UPEx) in human bone marrow mesenchymal stem cells (hBM-MSCs). UP-Ex protected hBM-MSCs against oxidative injury, as determined by MTT assays. This effect was confirmed by immunoblot analysis of the oxidation-sensitive protein p53 and the apoptotic protein cleaved caspase-3. Excessive intracellular reactive oxygen species (ROS) accumulation induced by oxidative stress was moderated in UP-Ex-treated hBM-MSCs (UP-Ex-MSCs). Similarly, expression of the ROSscavenging enzymes superoxide dismutase 1 (SOD1), SOD2, and catalase was recovered in UPEx- MSCs. Excessive ROS induced by long-term cell expansion (passage 17) was significantly decreased along with restoration of the senescence proteins p53, p21, and p16 in UP-Ex-MSCs. UP-Ex treatment also improved the ability of these replicative, senescent hBM-MSCs (passage 17) to differentiate into osteocytes or adipocytes, suggesting that UP-Ex ameliorates the functional decline of senescent stem cells and may provide better therapeutic efficacy in stem cell therapy.
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The Role of Stress Granules in the Neuronal Differentiation of Stem Cells
정신구,온탁범,Chul Ho Jang,Karthikeyan Vijayakumar,조광원 한국분자세포생물학회 2020 Molecules and cells Vol.43 No.10
Cells assemble stress granules (SGs) to protect their RNAs from exposure to harmful chemical reactions induced by environmental stress. These SGs release RNAs, which resume translation once the stress is relieved. During stem cell differentiation, gene expression is altered to allow cells to adopt various functional and morphological features necessary to differentiate. This process induces stress within a cell, and cells that cannot overcome this stress die. Here, we investigated the role of SGs in the progression of stem cell differentiation. SGs aggregated during the neuronal differentiation of human bone marrow-mesenchymal stem cells, and not in cell lines that could not undergo differentiation. SGs were observed between one and three hours post-induction; RNA translation was restrained at the same time. Immediately after disassembly of SGs, the expression of the neuronal marker neurofilament-M (NFM) gradually increased. Assembled SGs that persisted in cells were exposed to salubrinal, which inhibited the dephosphorylation of eukaryotic translation initiation factor 2 subunit 1 (eIF2α), and in eIF2α/S51D mutant cells. When eIF2α/S51A mutant cells differentiated, SGs were not assembled. In all experiments, the disruption of SGs was accompanied by delayed NF-M expression and the number of neuronally differentiated cells was decreased. Decreased differentiation was accompanied by decreased cell viability, indicating the necessity of SGs for preventing cell death during neuronal differentiation. Collectively, these results demonstrate the essential role of SGs during the neuronal differentiation of stem cells.
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홍조류인 Gracilaria vermiculophylla 추출물에 의한 노화 골수유래 중간엽줄기세포의 항노화 및 분화능력 개선 효과
정신구(Sin-Gu Jeong),조태오(Tae Oh Cho),조광원(Goang-Won Cho) 한국생명과학회 2018 생명과학회지 Vol.28 No.9
홍조류인 꼬물꼬시래기(Gracilaria vermiculophylla)는 전 세계의 해변 지역에 널리 퍼져 있으며 아시아 국가에서 식량 자원으로 이용되어왔다. 이전 연구에 따르면, Gracilaria 속 홍조류 추출물에서 항산화 및 항염증 효과가 보고되었다. 본 연구에서는 노화된 인간의 골수 유래 중간엽 줄기세포(hBM-MSCs)를 이용하여 Gracilaria vermiculophylla 추출물(GV-Ex)의 항노화 효과를 조사하였다. MTT 분석와 immunoblot 분석(apoptotic protein p53과 cleaved caspase-3)을 이용하여, GV-Ex 전처리는 산화적 스트레스에 의해 손상된 hBM-MSCs의 세포생존력을 향상시킴을 확인하였다. 또, 세포내 생성된 ROS는 장기간 배양 된 MSCs (Passages 17; P-17)와 P-7 MSC에서 측정하여 서로 비교하였는데, P-17 MSC에서 증가되었고, GV-Ex 처리하면(GV-Ex treated P-17 MSCs) 유의하게 감소되었다. 또한, 항산화 효소인 SOD1와 SOD2, CAT의 발현 역시 GV-Ex 처리함에 따라 복원됨을 관찰하였다. 노화표지단백질인 p53와 p21, p16 등의 발현 또한 GV-Ex를 처리한 P-17 MSC에서 감소되었다. 줄기세포의 골세포(osteocytes) 혹은 지방세포(adipocytes)로 분화하는 능력 역시 GV-Ex를 처리한 P-17 MSCs에서 개선되었다. 이상과 같은 결과를 통해, GV 추출물은 노화된 줄기세포의 기능을 개선함을 시사한다. The red algae Gracilaria vermiculophylla is widespread on seashores worldwide and has been used as food in Asian countries. Previous studies have reported that extracts of Gracilaria red algae have beneficial anti-oxidant and anti-inflammatory effects. The present study examined the anti-senescence effects of Gracilaria vermiculophylla extracts (GV-Ex) in replicatively senescent human bone marrow mesenchymal stem cells (hBM-MSCs). GV-Ex pretreatment improved the cellular viability of hBM-MSCs that had been injured by oxidative stress. These effects of GV-Ex were confirmed by MTT assay and immunoblot analysis using the apoptotic proteins p53 and cleaved caspase-3. The reactive oxygen species (ROS) levels were examined in long-term cultured Passages 17 (P-17) mesenchymal stem cells (MSC) and compared to P-7 MSC. The ROS accumulation was greater in the P-17 than in the P-7. However, these increased ROS levels in the P-17 were decreased significantly after treatment with GV-Ex, and restoration of the levels of the anti-oxidant enzymes SOD1, SOD2, and CAT was also observed under these conditions. In addition, P-17 hBM-MSC treated with GV-Ex had decreased levels of the senescence proteins p53, p21, and p16. The results show that the ability of P-17 hBM-MSC to differentiate into osteocytes and adipocytes was improved by GV-Ex treatment, suggesting that GV-Ex ameliorates the functional decline of senescent stem cells.
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Resveratrol 처리한 HeLa세포에서 angiogenin과 vascular endothelial growth factor 발현유도에 따른 세포이동촉진
조이슬(I-Seul Joe),정신구(Sin-Gu Jeong),조광원(Goang-Won Cho) 한국생명과학회 2014 생명과학회지 Vol.24 No.4
Resveratrol (RSV)은 천연 폴리페놀계 화합물로 세포분열, 성장, 세포이동 등과 같은 다양한 효과가 보고되었다. Angiogenin (ANG)은 Vascular endothelial growth factor (VEGF)와 함께 세포의 증식, 신생혈관형성, tubular structures의 형성, 세포이동 등을 이끄는 중요한 단백질이다. 본 연구에서는 RSV에 의한 세포이동효과를 HeLa 세포에서 관찰하였다. Real-time PCR을 통해 HeLa 세포에 RSV 0~50 μM을 24시간 동안 처리하였을 때, 농도에 따른 ANG, VEGF 유전자 발현이 의미 있게 증가 하였다. 같은 방법으로, RSV 50 μM을 시간에 따라(0~48시간)처리하여 실험하였다. 그 결과, RSV 50 μM을 24시간 동안 처리하였을 때 ANG, VEGF 유전자 발현이 가장 높게 증가하였고, ANG 단백질 분석에서도 동일한 결과를 얻었다. 또한, MTT assay를 이용한 세포 독성연구에서, RSV 50 μM의 농도에서는 영향을 미치지 않음을 관찰하여, 이를 최적의 조건으로 결정하였다. RSV가 처리된 세포에서 세포이동효과를 조사하기 위해 wound-healing assay를 수행하였다. RSV가 처리된 그룹에서 세포이동이 의미 있게 증가하였다. 따라서, 본 연구에는 RSV에 의해 ANG, VEGF의 발현이 증가했고, 이에 따라 세포이동이 향상됨을 관찰하였다. Resveratrol (RSV), a natural polyphenolic compound, is a modulator for cell division and cell migration, and has diverse beneficial properties. Angiogenin (ANG) and vascular endothelial growth factor (VEGF) are considered to be important mechanisms for cell proliferation, angiogenesis, the formation of tubular structures, and migration. In this study, we investigated whether RSV has a migratory effect in HeLa cells. When cells were treated with 0~50 μM of RSV for 24 hr, the expression of ANG and VEGF was significantly increased in a dose dependent manner measured by real-time PCR. Similarly, we performed time dependent experiments for 50 μM RSV treated cells and identified the optimized time at 24 hr. The increased expression in RSV treated cells was confirmed by Western blot analysis. To examine the toxic effects of RSV at the determined conditions, MTT assays were performed. The viabilities were unchanged for 0~50 μM RSV treated cells, while they decreased at 100 μM RSV. To examine the effect of migration in RSV treated cells, we performed a wound-healing assay. The migratory rates were significantly enhanced in the RSV treated group. In this study, we found that RSV induces an increase in the expression of migration factors ANG, VEGF, and enhances cell migration for the determined conditions.
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