臺灣水鹿鹿茸於傷口癒合及呼吸道發炎小鼠之研究

Abstract

鹿茸為傳統東方藥學中之名貴藥材，於過去研究中指出茸角週期與鹿茸內部多種生長因子相關，且鹿茸之生長因子亦已知能助於皮膚之傷口癒合；此外，於本實驗室先前研究已證實臺灣水鹿鹿茸具有抗感染與抗過敏之效果；因此，本研究針對臺灣水鹿鹿茸之鹿茸粉與其萃取物對於促進傷口癒合及抗氣喘進行探討。 於細胞實驗中，將鹿茸冷水與熱水萃取粉和人類正常角質細胞株（HaCaT）共同培養後，再分別測定細胞增生與細胞遷移；結果顯示，鹿茸冷水與熱水萃取粉皆可顯著增加人類皮膚角質細胞增生，且鹿茸冷水萃取粉能夠顯著增加角質細胞遷移；另以鹿茸冷水與熱水萃取粉與人類纖維母細胞（CCD-966SK）共同培養，結果顯示，鹿茸冷水與熱水萃取粉皆不顯著影響纖維母細胞之增生。而將鹿茸粉與鹿茸冷水萃取粉運用於傷口癒合動物實驗中，結果顯示，鹿茸粉與鹿茸冷水萃取粉對於小鼠皮膚傷口之癒合皆無顯著改善。 運用鹿茸粉全程餵食雞卵蛋白誘導之氣喘小鼠，可降低氣喘小鼠支氣管沖洗液中免疫細胞聚集，顯著降低第二型輔助性T細胞之細胞激素（介白素-4、介白素-5與介白素-13）與前發炎反應細胞激素（介白素-1β與C-C motif ligand 20）之含量，改善支氣管週邊細胞浸潤與呼吸道壁之發炎反應，降低由不同濃度乙醯甲膽鹼引起之呼吸道過度反應；另一方面，鹿茸粉對於氣喘小鼠呼吸道中黏液分泌與血清中免疫球蛋白之抗體含量則無顯著差異。 結果顯示，臺灣水鹿鹿茸之萃取物可促進角質細胞之增生與遷移，而鹿茸粉能夠改善氣喘小鼠之呼吸道過度反應；對於影響傷口癒合與氣喘之可能機制需要再做深入的研究，且對其內部之有效成分做進一步之探討。

Velvet antler (VA) is one of the most famous and valuable Chinese traditional medicines. Several studies indicated that the VA comprises certain growth factors related to its regeneration annually. These growth factors might provide positive effects on wound repairs. In addition, in our previous study, we also demonstrated the anti-allergic and anti-inflammatory effects of VA from Formosa sambar deer (Rusa unicolor swinhoii). Thus, the purpose of this study was to further evaluate the effects of VA and its extracts (VA cold-water-soaked and VA water-boiled extracts) from Formosan sambar deer on the wound healing and anti-asthmatic effects in vitro and in vivo. For wound healing study, the human keratinocyte cells (HaCaT) were co-incubated with VA extracts. Results indicated that both VA cold-water-soaked (VACWS) and VA water-boiled-extract (VAWB) samples could stimulate keratinocyte proliferation in a dosage-dependent manner. VACWS were also demonstrated a significantly higher wound repairing ability than other samples in a dosage-dependent manner using cell migration assay. On the other hand, the VA extracts were co-incubated with human fibroblast cells (CCD-966SK). The results indicated that both VA cold-water-soaked (VACWS) and VA water-boiled-extract (VAWB) samples have no significant effect on cell proliferation. The wound healing animal model did not show any significant difference among treatments in wound healing. For anti-asthmatic study, the asthmatic mice induced by ovalbumin were used to evaluate the anti-hyper-responsive effect of the VA powder. The results clearly demonstrated that the VA powder has an anti-asthma effect with ameliorating the symptom of airway hyper-responsiveness (AHR). The levels of Type 2 helper T cell (Th2) cytokines, such as interleukin (IL)-4, IL-5 and IL-13, in brochoalveolar lavage (BAL) were significantly decreased. The total eosinophil count induced by ovalbumin in BAL was also reduced. Interestingly, OVA-specific IgE in mice serum was not reduced by the VA treatments. In summary, these findings clearly revealed that VACWS has potential for wound healing and the VA powder could ameliorate the symptom of AHR. Further studies are necessary to reveal the mechanisms and to determine the possible compounds involved in the wound repairs and anti-allergic effects.