부산대학교 도서관

연구목적: 혈관 평활근세포 증식에 대한 국내 자생종 승마의 효과를 밝히기 위해 시행되었다. 연구재료 및 방법: 3개월된 암컷 Sprague-Dawley 쥐의 대동맥으로부터 혈관 평활근세포를 얻었다. 눈빛승마, 왜승마, 세잎승마, 촛대승마 등 4종의 자생 승마를 80% ethanol, absolute methanol 그리고 40% 2-propanol을 사용하여 추출하였다. 10% fetal bovine serum (FBS)에서 배양한 혈관 평활근세포를 승마 추출물 (0.3mg/mL)로 24시간 동안 처리한 후 bromodeoxyuridine incorporation assay를 사용하여 세포증식을 조사하였다. dichlorofluorecin diacetate을 첨가한 후 flow cytometry를 이용하여 세포내 ROS 생성을 정량화하였다. 결과: 추출법이 다른 국내 자생종 승마 추출물을 처리한 결과, 혈관 평활근세포의 증식이 유의하게 23-30% 억제되었다. 또한 강력한 항산화제인 비타민 E는 10% FBS로 유도된 세포증식을 감소시켰다. 승마 추출물의 작용기전을 밝히기 위한 추가 연구 상 10% FBS에 의한 ROS 생성이 승마에 의해 감소되었다. 세포증식이나 ROS 생성에 대한 효과는 추출 방법과 승마 종에 따라 차이가 없었다. 결론: 국내 자생종 승마는 산화 스트레스 감소를 통하여 혈관 평활근세포 증식을 억제한다. 향후 다양한 심혈관계 영향에 대한 추가 연구가 필요할 것으로 사료된다. Objectives: To examine the effect of extracts of Korean native Cimicifuge species on cell proliferation in vascular smooth muscle cells (VSMC).
Methods: VSMC were isolated from rat aorta. Cell proliferation was assessed by measure of bromodeoxyuridine incorporation into the cells. Differences in Reactive oxygen species (ROS) levels were examined after exposure to the extracts of Korean native Cimicifuge species using the detection reagents dichlorofluorecin diacetate. The rhizomes/roots were air-dried and milled with a commercial food mixer. Milled rhizomes/roots of each Cimicifuga species were separately extracted by 80% ethanol, absolute methanol, and 40% 2-propanol using homogenizer and evaporated under reduced pressure at low temperatures. Effects of extracts dissolved in phosphate-buffered saline (0.3 mg/mL) were examined. Results: Ethanolic, methanolic or propanolic extracts of 4 Korean native Cimicifuge species (Cimicifuga [C] davurica, C. japonica, C. heracleifolia var. bifida Nakai, C. simplex) were screened. The addition of extracts of each Korean native Cimicifuge species to cells in the presence of 10% fetal bovine serum (FBS) potently inhibited cell proliferation. Significant decrease of 23%-30% was observed. Vitamin E, a potent antioxidant, inhibited 10% FBS-stimulated cell proliferation of VSMC. We also demonstrated that extracts of each Korean native Cimicifuge species decreased intracellular ROS generation induced with 10% FBS. The effect of Korean native Cimicifuge species was not species-specific and solvent-specific. Conclusion: TExtracts of Korean native Cimicifuge species inhibit VSMC proliferation via inhibition of intracellular ROS. These findings suggest that Cimicifuge species used for reducing menopause symptoms might be cardioprotective in women.