본 연구팀에서는 방사선으로부터 위장관과 면역조혈계를 방호하기 위하여 당귀, 천궁, 백작약으로 새로운 생약복합물 HIM-I을 개발하였고, HIM-I을 에탄올 침전하여 에탄올 분획(HIM-I-E)과 조다당 분획(HIM-I-P)을 얻은 후, HIM-I에 조다당 분획을 첨가하여 HemoHIM을 제조하여 실험하였으며, 과산화수소를 이용하여 산화적 스트레스를 유발한 후 HemoHIM의 apoptosis 억제 효과를 측정하였다. 과산화수소를 세포주에 처리한 결과 세포독성 증가, 세포막 파괴, subG1기 세포 증가, DNA 절단 현상 등이 관찰되었다. 이때 HIM-I, HIM-I-P, HemoHIM을 첨가하여 전처리 배양한 다음에 과산화수소로 apoptosis를 유도하였다. 그 결과 과산화수소에 의한 세포독성, 세포막 파괴, subG1기 세포, DNA 절단현상 등이 현저하게 감소하는 것으로 나타났다. 전반적으로 HemoHIM의 방호 효과는 HIM-I-P와 비슷하였으며 HIM-I보다 높은 것으로 나타났다. 이상의 결과로 HemoHIM은 면역기능 방호 회복 물질로서 뿐만 아니라 산화적 스트레스에 대한 항산화제로서 유용하게 활용될 수 있을 것으로 생각된다.
In our previous study, a novel herb mixture (HIM-I) of Angelica gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix was developed to protect the intestinal and immune systems and promote its recovery against radiation damage. A new herbal composition (HemoHIM) with the high immune modulating activity was developed from HIM-I. HIM-I was fractionated into ethanol fraction (HIM-I-E) and polysaccharide fraction (HIM-I-P). And HemoHIM was prepared by adding HIM-I-P to HIM-I. HemoHIM showed more effective than HIM-I in immune modulation as well as radioprotection. The present study is designed to investigate the protective effects of HIM-I, HIM-I-P, and HemoHIM on hydrogen peroxide $(H_2O_2)$ induced apoptosis of human promyelocytic leukemia (HL-60) cells. It was shown that $H_2O_2$ treatment reduced the viability of cells, and increased appearance of DNA ladders, hypodiploid (subG1) cells, and phosphatidylserine translocation level. Pretreatment of HemoHIM significantly reduced the cytotoxic effect induced by $H_2O_2$, associated with reducing the translocation of phosphatidylserine, hypodiploid cells and DNA ladders. HemoHIM appeared to be more protective than HIM-I against $H_2O_2$ induced apoptosis whereas, it exhibited similar activity to HIM-I-P. These results indicated that HemoHIM might be an useful agent for protection against oxidative stress $(H_2O_2)-induced$ apoptosis as well as immune modulation, especially since it is a relatively nontoxic natural product.
