Effect of Cervi Pantotrichum Cornu Herbal acupuncture on protease activities, antioxidant in Rheumatoid arthritis rats

류마티스 관절염 실험용쥐의 활액에서 단백분해효소의 활성 및 항산화에 대한 녹용약침의 효과

We have compared(using the same series of experimental tissue samples) the levels of proteolytic enzyme activities and free radical-induced protein damage in synovial fluid from RA and CPH cases. Many protease types showed significantly increased (typically by a factor of approximately 2-3-fold) activity in RA, compared to normal rats. However, CPH significantly reduced the cytoplasmic enzyme activities of arginyl aminopeptidase, leucyl aminopeptidase, pyroglutamyl aminopeptidase, tripeptidyl aminopeptidase, and proline endopeptidase to almost about 1/10 each. For the Iysosomal proteases, synovial fluid samples from RA rats, CPH significantly reduced the enzyme activities of cathepsin B, dipeptidyl aminopeptidase I and dipeptidyl aminopeptidase II. In extracellular matrix degrading(collagenase, tissue elastase) and leukocyte as sociated proteases (leukocyte elastase, cathepsin G), CPH decreased these enzyme activities of collagenase, tissue elastase and leukocyte associated elastase in RA. In cytoplasmic and lysosomal protease activities in plasma from RA. CPH and normal plasma samples were not significantly different, suggesting that altered activity of plasma proteases (particularly those enzymes putatively involved in the immune response) is not a contributory factor in the pathogenesis of RA. In addition, the level of free radical induced damage to synovial fluid proteins was approximately twice that in RA, compared with CPH. CPH significantly decreased the level of ROS induced oxidative damage to synovial fluid proteins (quantified as protein carbonyl derivative). Therefore we conclude that both proteolytic enzymes and free radicals are likely to be of equal potential importance as damaging agents in the pathogenesis of inflammatory joint disease, and that the design of novel therapeutic strategies for patients with the latter disorder should include both protease inhibitory and free radical scavenging elements. In addition, the protease inhibitory element should be designed to inhibit the action of a broad range of protease mechanistic types (i.e. cysteine-, metallo- and serine- proteinases and peptidases). However, increased protein damage induced by ROS could not be rationalised in terms of compromised antioxidant total capacity, since the latter was not significantly altered in RA synovial fluid or plasma compared with CPH.

류마티스 관절염의 쥐의 활액에서 단백분해효소, 산화제와 유리기에 대한 녹용약침의 비특이적 면역억제효과를 연구하였다. 일련의 실험표본으로서 여러가지 세포질, 리소좀, 기질 백분해효소의 제 활성을 RA대조군과 녹용약침군의 활액에서 카르보닐기 유도로 생성되는 유리기-유발 단백질손상과 항산화를 비교하였다. 전반적으로 단백분해효소활성이 정상군과 비교하여 RA대조군에서 유의성 있게 증가하였다. 세포질 단백분해효소들은 정상군과 RA군의 차이에서는 유의성이 없었다. 녹용약침처리($100{\mu}g/kg$)결과 세포질, 리소좀, 기질 단백분해효소생성을 억제하였으며, RA군과 녹용약침군 또는 정산군 사이에 활액 또는 세포질 항산화에서 유의성 있는 차이가 없음에도 불구하고, RA군 활액의 단백질손상을 유발하는 유리기는 녹용약침군과 정산군에 비교하여 약 2배 정도 높았다. 이상의 결과에서 단백분해효소와 유리기는 RA유발시 단백질손상을 유도하는 물질로 밝혀졌으며, 따라서 단백분해효소 저해와 유리기소거능을 갖는 치료법개발이 새로운 RA예방치료법으로 제시되었다. 나아가서 여러가지 기질특이성을 갖는 활액내 단백분해효소류(cysteine, serine, metallo proteinases와 peptidases)에 대한 효과적인 저해제개발이 필요한 것으로 보인다. 따라서 본 녹용약침은 이와 같은 새로운 개념의 2가지(유리기제거, 단백분해활성) 관절염치료 요소를 충족하는 약리활성을 포함하는 훌륭한 제제로 평가된다.