The tonic effect and available components of $\underline{Panax}$ $\underline{Ginseng}$ $\underline{Alaliaceae}$ was studied $\underline{in}$ $\underline{vitro}$ by a new biochemical approach employing a rabbit skeletal muscle enzyme, Creatine phosphokinase(CPKase, EC2.7.3.2), which catalyzes the reversible transphosphrylation between ATP and creatine molecules. Under the standard experimental condition, the activity of CPKase was measured by the addition of ginseng extracts, including the known bioactive components of Panax Saponins, which were fractionated by organic solvent extraction method and further by DEAE cellulose ion exchange chromatography. The results are as follows;
1. In the presence of Panax Saponin A (PSA), and Panax Saponin C (PSC), the activity of CPKase was relatively increased. However, the activity lowered when the amount of PSA rised over 1 mg, but in case of PSC, the enzyme activity was constant value at about 1.3 times of control.
2. Ginseng ether extract (G-Eth) was observed to inhibit CPKase activity. The inhibition degree was decreased with the increase of G-Eth amount added, and at 2mg of G-Eth, the enzyme activity decreased to almost half.
3. The CPKase activity was increased gradually as the amount of Ginseng methanol extracts added. It was found that, however, the enzyme activity was decreased when the amount of Ginseng methanol extract (G-Met-1) was over 7mg, but when G-Meg-2 which is decolorized G-Met-1 by active carbon, was constant value at about 1.8 times of control system.
4. G-Met-2 was further fractionated by DEAE cellulose column chromatography and three fractions(Fr I, Fr II, Fr III) was obtained. The activity of CPKase was increased gradually as the amounts of FrI added. In case of FrII and FrIII, the enzyme activity was greatly increased in the presence of small amount such as 0.1mg, but the activity was decreased quickly when the amount of FrII and FrIII was over 0.1mg. At 0.5mg of FrII and 3.0mg of FrIII, the activity was equal to control condition and at above the amount, it was found that CPkase activity was inhibited.
5. It was observed that FrI containe saponin, mono and di saccharide, and peptides, but no information was obtained about FrII and FrIII.
인삼의 약효 및 생리작용의 본질을 밝히고 인삼유효성분을 분리하기 위한 새로운 생화학적 시도로 토끼골격근 Creatine Phosphokinase 를 이용하여 $\underline{in vitro}$에서 연구한 결과:
1) Panaxatriol 계 saponin인 Panax Saponin A 및 Panax Saponin C는 CPKase활성을 증가시켰으며, PSA의경우 1mg이상에서는 오히려 활성증가의 감소가 일어나 5mg첨가시 거의 대조군과 비슷한 활성을 보였고, PSC는 1mg이상에서 거의 1.31배 정도의 활성증가를 유지하였다.
2) 인삼 ether 추출물 (G - Eth) 첨가시는 CPKase 활성이 억제 되었으며, G - Eth 첨가량의 증가에 따라 활성억제도는감소되었고, 2mg 첨가시는 대조군에 비해 거의 반정도의활성밖에 보이지 않았다.
3) 인삼 methanol 추출물 (G-Met) 첨가시 G-Met-1, G-Met2 공히 첨가량의 증가에 따라 급격한 활성증가를 보이다가 7mg이상이 되면 G-Met-1은 오히려 활성의 감소를 보인반면, G-Met-2는 거의 1.8배 정도의 일정한 활성증가를 유지하였다.
4) G-Met-2 를 DEAE cellulose ion exchange chromatography에 의해 분리한 결과 CPKase의 활성을 증가시키는 세분획을 얻었으며 (FrI, FrII, FrIII), 각분획에 대한 CPKase활성을 조사해본 결과 FrI은 첨가량의 증가에 따라거의 비례하여 활성이 증가됨을 알 수 있었고, FrII 및 FrIII는 0.1mg정도의 소량첨가시는 상당한 활성의 증가를 보였으나, 그 이상이 되면 급격한 활성의 감소를 보여 FrII의 경우 0.5mg 첨가시, FrII는 3.0mg 첨가시 대조군과 거의 비슷한 활성을 보였고, 그이상이 되면 오히려 CPKase 활성을 억제하였다.
5) FrI 은 saponin을 비롯한 당류, peptide 등이 포함되어 있음을 알았으나, FrII 및 FrIII는 어떤 계통을 물질인지 아직 밝혀지지 않았다.
