진세노사이드 Rb1과 Rg1에 의한 해마 신경전구세포의 분화 증가 Ginsenosides Rb1 and Rg1 Decrease Proliferation but Increase Neuronal Differentiation of Hippocampal Neural Progenitor Cells원문보기
전세계적으로 널리 복용되고 있음에도 불구하고 인삼이 중추신경계에 미치는 효과에 대한 세포수준에서의 증거는 별로 없다. 따라서 본 연구자들은 지금까지 보고된 30여종 이상의 ginsenosides 중에서 인삼 효과의 주된 활성성분으로 알려져 있는 Rb1과 Rg1을 이용해 해마신경전구세포의 분화와 증식에 미치는 효과를 연구하였다. 증식실험결과 Rb1과 Rg1을 3일 동안 해마신경전구세포에 처리하면 대조군에 비해 BrdU(+)세포수가 상당히 감소한 반면에 분화조건에서 Rb1과 Rg1을 처리했을 때는 신경세포특이적인 MAP2 단백질을 발현하는 세포수가 증가하였다. 전구세포의 신경세포로의 분화 결정에 관여한다고 잘 알려져 있는 proneural 전사인자인 Ngn1과 Hes1 유전자의 발현양상도 대조군에 비해 증가하였다. 따라서 본 연구 결과, ginsenoside Rb1과 Rg1은 해마신경전구세포의 증식을 감소시키고 대신 신경세포로의 분화를 촉진시킴으로써 해마신경발생에 관여함을 보여준다.
전세계적으로 널리 복용되고 있음에도 불구하고 인삼이 중추신경계에 미치는 효과에 대한 세포수준에서의 증거는 별로 없다. 따라서 본 연구자들은 지금까지 보고된 30여종 이상의 ginsenosides 중에서 인삼 효과의 주된 활성성분으로 알려져 있는 Rb1과 Rg1을 이용해 해마신경전구세포의 분화와 증식에 미치는 효과를 연구하였다. 증식실험결과 Rb1과 Rg1을 3일 동안 해마신경전구세포에 처리하면 대조군에 비해 BrdU(+)세포수가 상당히 감소한 반면에 분화조건에서 Rb1과 Rg1을 처리했을 때는 신경세포특이적인 MAP2 단백질을 발현하는 세포수가 증가하였다. 전구세포의 신경세포로의 분화 결정에 관여한다고 잘 알려져 있는 proneural 전사인자인 Ngn1과 Hes1 유전자의 발현양상도 대조군에 비해 증가하였다. 따라서 본 연구 결과, ginsenoside Rb1과 Rg1은 해마신경전구세포의 증식을 감소시키고 대신 신경세포로의 분화를 촉진시킴으로써 해마신경발생에 관여함을 보여준다.
Ginseng is the best known and most popular herbal medicine used worldwide. In spite of reported beneficial effects of ginseng on the CNS, there is few scientific evidences established at the cellular level. Among more than 30 ginsenosides, Rb1 and Rg1, the active ingredients of ginseng, are regarded...
Ginseng is the best known and most popular herbal medicine used worldwide. In spite of reported beneficial effects of ginseng on the CNS, there is few scientific evidences established at the cellular level. Among more than 30 ginsenosides, Rb1 and Rg1, the active ingredients of ginseng, are regarded as the main compounds responsible for many pharmaceutical actions of ginseng. Daily treatment with Rb1 or Rg1 for 3 d significantly decreased the number of bromodeoxyuridine(BrdU)(+) cells in primary neural progenitor cells(NPCs) isolated from hippocampi at embryonic day 16.5(E16.5). In contrast, treatment with Rb1 or Rg1 greatly increased the number of microtubule associated protein(MAP2) (+) cells. In addition, the transcription factors, Ngn1 and Hes1, proneural members of the basic helix-loop-helix(bHLH) family, significantly increased in Rb1 or Rg1 treated-NPCs. Based on these results, we suggest for the first time that ginsenosides Rb1 and Rg1 decrease proliferation but promote neuronal differentiation of hippocampal NPCs.
Ginseng is the best known and most popular herbal medicine used worldwide. In spite of reported beneficial effects of ginseng on the CNS, there is few scientific evidences established at the cellular level. Among more than 30 ginsenosides, Rb1 and Rg1, the active ingredients of ginseng, are regarded as the main compounds responsible for many pharmaceutical actions of ginseng. Daily treatment with Rb1 or Rg1 for 3 d significantly decreased the number of bromodeoxyuridine(BrdU)(+) cells in primary neural progenitor cells(NPCs) isolated from hippocampi at embryonic day 16.5(E16.5). In contrast, treatment with Rb1 or Rg1 greatly increased the number of microtubule associated protein(MAP2) (+) cells. In addition, the transcription factors, Ngn1 and Hes1, proneural members of the basic helix-loop-helix(bHLH) family, significantly increased in Rb1 or Rg1 treated-NPCs. Based on these results, we suggest for the first time that ginsenosides Rb1 and Rg1 decrease proliferation but promote neuronal differentiation of hippocampal NPCs.
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가설 설정
6 DIV cultures were exposed daily to Rbl or Rgl(50 and 100μM) for 3 d, followed by a 1 h pulse of BrdU(5()μM), and fixed on 9 DIV. B: Effects of different doses of Rbl or Rgl on proliferation. C: Apoptosis assay: green fluorescent nuclei correspond to apoptotic cells in which the tenninal transferase has incorporated fluorescent dUTP(Bodipy-dUTP) into the fragmented DNA.
Next, we hypothesized that Rbl and Rgl may influence the balance between the proliferation and the differen-tiation of NPCs. To address this possibility, Pl progenitors expanded in N2 in the presence of bFGF were rinsed free of bFGF and then, treated with Rbl or Rgl for 3 din NB medium to allow differentiation.
제안 방법
For high-density cultures, the cells on 80% confluency were subcultured at a density of 6×104 cells/cm2 in a 24-well plate in N2 me-dium in the presence of bFGF and these subcultured cells were designated as passaged once(Pl). All experiments were carried out using the Pl neural progenitors. The neu-ral progenitors were induced to differentiate by with drawing bFGF and kept in a differentiation media(Neuro-basal medium: NB) for 3~5 d in the presence or absence of Rbl/Rgl.
, 2000), and the re-sulting cultures were characterized. To investigate whether Rbl and Rgl affected the proliferation of hippocampal NPCs, the cells were pulsed with BrdU(10μM), a marker for cell proliferation, during expansion. When pulsed with BrdU for 1 h 4 days in vz7ro(DIV) after passage, an aver-age 48.
데이터처리
The normalized expression values for all control and treated samples was averaged, and average fold changes were determined. A Student's t test was conducted be-tween the normalized relative expression values for each individual control and treated sanpies to determine stat-istical relevance.
Statistical significance was determined using Student's t-test. All statistical analyses were conducted with Sigma plot 2000 software.
후속연구
These observations led us to pro-pose that Rgl, via the activation of Ngnl, might play an instructive role which on the one hand induces neuronal differentation and on the other hand inhibits the production of glial cells. Further work needs to be done to determine how ginsenosides regulate the glial cell differentiation and the expression of bHLH genes required for neuronal differ-entiation of NPCs.
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