- 學位論文
北冬蟲夏草無菌栽培研究及動物實驗
The Culture of Cordyceps militaris and Animal Experiments
摘要
冬蟲夏草 (Cordyceps sinensis) 自古有云「冬蟲夏草、甘平、保肺益腎、止血化痰、止勞咳…」，《中醫藥典》認為產於青康藏高原的冬蟲夏草確有改善呼吸道疾患之功效，因其稀少不易取得，價格居高不下。北冬蟲夏草 (Cordyceps militaris) 又名北蟲草或蛹蟲草，是子囊菌亞門，麥角菌目，麥角菌科，蟲草屬的模式種，有著與天然冬蟲夏草同樣的生活史，與Cordyceps sinensis極其近似，是冬蟲夏草唯一的替代品，但在自然界北蟲草亦是一種罕見而難以採收的草藥，本實驗室取天然成熟北蟲草之子實體利用人工培育的方式成功地培養出子實體，約七十天即可成熟採收。接著進行動物實驗，利用實驗室採收之北蟲草子實體，首先進行急性毒性及慢性毒性動物實驗，根據實驗中之小白鼠均未有死亡現象，並於犧牲解剖後檢查所有組織正常，證明北蟲草並不具有毒性。 由於北蟲草與冬蟲夏草性質相近，我們想要證實北蟲草是否對於呼吸道緩解具有療效，因而進行人工誘發氣喘之動物試驗。在急性氣喘試驗中，我們以蒸餾水作為陰性對照組；麻杏甘石湯作為陽性對照組；北蟲草液為實驗組，各液體進行腹腔注射後誘發氣喘，結果發現:注射麻杏甘石湯及北蟲草液的小鼠，對於氣喘發作具有較長時間的耐受性，注射蒸餾水平均耐受性為 96.6±7.6秒；注射麻杏石甘湯為172.8±37.4秒；注射北蟲草液1ml、2ml與3ml其耐受性分別為174.4±25.0；177.6±25.1 及 196.0±8.9秒 (P<0.05) 。在長期餵食北蟲草之氣喘試驗中，將6周齡之BALB/c小鼠餵食北蟲草滿四週後，再使用吸入性組織胺引發氣喘反應，最後犧牲小鼠取其氣管作病理切片及細胞計數，結果發現無餵食北蟲草之對照組小鼠的氣管徑四隻中有三隻明顯收縮變形；而有餵食北蟲草的實驗組的四隻小鼠之氣管均無明顯收縮現象。鏡檢發現，無餵食北蟲草之小鼠的嗜伊紅性白血球有明顯增多 (>5/HPF) 而有餵食組平均為2/HPF (P<0.05) 。由以上兩組動物實驗清楚顯示餵食北蟲草後之實驗組動物對人工誘發之氣喘均可產生緩解的功能。所以我們已經可以成功地人工培養出北蟲草子實體，也可以從實驗中得知北蟲草對於氣喘確實具有改善之功效也可以印證中醫藥典上對於北蟲草功效的論述。
並列摘要
Cordyceps belongs to the family of numerous mushrooms, which is, actually, parasitic fungi, growing wildly on the caterpillars in the high plateaus of China, Nepal, and Tibet. It has the ability to enhances the immune system and improves the lung function from the ancestral codex. But Cordyceps sinensis are rare and difficult to collect, most of the available commercial products belong to the mycelium production, since the real mature mechanism are in the carpophore stage. According to the official published Chinese medicine book, the only substitute herb is Cordyceps militaris. We have developed a cultivating method in laboratory for the growing of the carpophore stage of C. militaris. Several papers have been proved the anti-oxidative function of C. militaris and some traditional medicine researches also mention the improvement of the human asthma condition. In this study, we have done the acute and chronic C. militaris toxicity tests for the mice and could not find out any adversities nor any side effects from the results. In acute asthma test, we used the extract of C. militaris to inject into the peritoneum of the mice and use the Ma Hsing Kan Shin Tang (M.H.K.S.T) injected mice as the positive control. The results have been shown the C. militaris injected mice and the positive control groups which have the longer ability to stand with the histamine challenge, if they compare with the normal saline injected mice (normal saline injected mice: 96.6±7.6 seconds; M.H.K.S.T mice : 172.8±37.4 seconds; C. militaris mice: 1ml- 174.4±25.0 seconds; 2ml- 177.6±25.1 seconds and 3ml- 196.0±8.9 seconds (P<0.05) . And we also fed C. militaris exrtract for the mice a period and sensitized with ova-albumin at the same time, then compared with the control group (non-sensitized mice) to observe the effects of C. militaris to the airway hypersensitivity. The result showed the sensitized mice without C. militaris feeding have prominent airway contraction about 75%; however, all sensitized mice feeding with C. militaris had no airway constriction. Furthermore, the sensitized mice without extract feeding have found more eosinophil counted under the high power field microscopy (>5-6/HPF) compared with the extract feeding mice (2/HPF) (P<0.05). In conclusion, these experiments were shown the laboratory cultured C. militaris has improved the asthma and airway condition in tested animals, that’s to says in the future we still need to do more extensive researches.