桂枝가 Angiogenesis의 抑制機轉에 미치는 영향 = Study on the Effect of Cinnamomi Ramulus(桂枝) on angiogenic inhibition mechanism

국문 초록 (Abstract)

桂枝의 apoptosis 유도와 angiogenesis 억제 기전을 실험적으로 규명하고자 ECV304 cell line 과 ECVPAR cell line을 이용하여 실험을 시행하여 다음과 같은 결과를 얻었다. 첫째, 세포부착 능력의 경우, 桂枝 煎湯液은 EVC304 cell line 과 ECVPAR cell line에서 5㎍/㎖부터 강한 저애효과를 보였다. 둘째, 桂枝 煎湯液은 ECV304 cell line서 GO/G1 arrest peak를 유발시켰다. 셋째, 桂枝 煎湯液은 ECV304 cell line에서 collagen coated plate에서 부착저지 효과를 나타냈다. 넷째, 桂枝 煎湯液은 ECV304 cell line에서 matrigel을 coating한 plate에서 lumen형성을 저애하였다. 다섯째, 桂枝 煎湯液은 ECV304 cell line 과 ECVPAR cell line에 대한 LFA-1 및 ELAM-1의 발현을 저애하였다. 여섯째, 桂枝 煎湯液은 ECV304 cell line 과 ECVPAR cell line에서 MMP-9과 uPA의 발현을 저애하였다. 일곱째, 桂枝 煎湯液은 ECV304 cell line 과 ECVPAR cell line에서 integrin αvβ3의 발현을 저애하였다. 이상의 실험결과로 보아 桂枝이 세포의 apoptosis를 유도하고 angiogenesis를 억제하는 효과가 있음을 알 수 있었으며, 또한 活血之劑의 angiogenesis 억제효과에 대한 가능성을 제시할 수 있을 것으로 사료된다.

다국어 초록 (Multilingual Abstract)

This experimental study was carried out to evaluate the effects of Cinnamomi Ramulus(CR) on angiogenic inhibition mechanism. In order to investigate the effects of Cinnamomi Ramulus on angiogenic inhibition mechanism evaluate cell survival rate by MTT assay, cell adhesive inhibition effect, DNA fragmantaion analysis, Nuclear Condensation assay, FACScan analysis, Angiogenesis inhibition assay, lmmunocytochemistry analysis, RT-PCR for mRNA expression, Western Blot analysis.
The results were summurized as follows :
1. The cell adhesive inhibition ability was strong from 5㎍/㎖.
2. The GO/G1 arrest peak was existed on ECV304 cell-line.
3. The cells on Collagen plate were inhibition of proliferation and inducement of apoptosis by CR water extract.
4. lumen formation was inhibited by CR water extract.
5. LFA-1 and ELAM-1's expression were inhibited by CR water extract. they are
commenly participation on inflammation and tumor regeneration.
6. The expression of MMP-9 and uPA were inhibited by CR water extract.
7. The expression of integrin αvβ3 was inhibited by CR water extract.
8. The expression of intracellular molecule were successively inhibited by CR water extract therefore the proliferation of ECV304 cell line was stopped and apoptosis was induced.
According to the results Cinnamomi Ramulus showed to be a key antagonist of
intergrin αvβ3, and which seems to be apoptosis by p53 through Flow cytometry. This report demonstrated that expression of MMP-9 and uPA blocked to under angiogenesis model. Thus, we suggested that Cinnamomi Ramulus block angiongenesis by inducing unscheduled apoptosis in ECV304 and ECVPAR cell lines and another oriental herbal medicine that treat Blood-stasis type also have angiogenic inhibition effects.