98.12.10第96130689號專利說明書及申請專利範圍修正本 九、發明說明： 【發明所屬之技術領域】 本發明係關於一種促進神經膠瘤細胞產生活性氧化物 之方法，特別是關於將至少含有香芹酚（carvacr〇1)之促 進液施予一神經膠瘤細胞，使該神經膠瘤細胞於外界微生物 入侵時，可提升活性氧化物之產生量 '使用裕度及細胞免 疫力之促進細胞產生活性氧化物之方法。 【先前技術】 一般而έ，一生物體中之細胞由於外界各種微生物（ mi咖rganism)之侵入’使得該生物體之細胞容易受該微生物 影響，進而造成該生物體之細胞的損害並導致該生物體產生病 香序紛由於具有抗S及殺g效果，因此已應用於殺除特 仅病菌及微生物，舉例而言，—習用殺菌劑，如中華民國專 利弟Π28975號「使用作為殺菌劑之含香_及麝香草驗之 發日轉利所示，其係包含香㈣及麝香轉，該香 二舌分係為5P，至9〇%乾重，該麝香草酚係為5ppm至80 j重’且該香_對麝香草盼之重量比係為1 : 5至i〇 : i 療由用於飲食組成物、飲水補充物或浸浴補充物中，以治 ^累旋體屬（Treponema)所造成之疾病，如_疾等。 )，為呈士生i勿粗内之活性氧化物（reactive⑽顺1 sPecies，ROS ，該糾.向度氧化力的分子’當細胞内的活性氧化物過量時 社法修魏物會攻擊醜、蛋自質及細趣㈣，進而造 成‘，，、去修㈣財，細，騎性氧化物對生命而言亦是必須 1321469 98. 12. 10第96130689號專利說明書及申請專利範圍修正本 的，例如超氧化物陰離子（㈣瓶ide anion),則是白血玻用 以^付病«之衫，因此，若生_並 生 ===氧化物絲持她可；反之，若峨= ^入該生物體内，則體内較佳係適當觸發該活性氧化物產生， 進而促進該生物體内之免疫功能。 …般而s，上述習用殺菌劑具有下列缺點，例如：該 =殺菌·剌於由密螺旋則所造成之疾病，進而造成 使用裕度储之缝；再者，該㈣ 物，並未提供-條峨使== 體内的活性氧化物產生量可能較為低落。基於上述原因，有 必要進一步改良上述習用殺菌劑。 ^鑑於此，本發明改良上述之缺點，其係經由將至少 t有t芽齡之促進液施予一神經膠瘤細胞，使該神經膠瘤細 胞於外界微，物=侵時’可提升該神經膠瘤細胞内活性氧 化物之產生量’藉此’本發明確實可提升使用裕度、活性 氧化物產生量及細胞免疫力。 【發明内容】 本發明主要目的係提供一種促進神經膠瘤細胞產生活 性氧化物之方法，其係經由將至少含有香芽紛之促進液施 予-神經膠瘤細胞，使該神經膠瘤細胞於外界微生物入侵時 ，可提升該神經膠瘤細胞内活性氧化物之產生量，使得本 發明具有提升使用裕度、活性氧化物產生量及細胞免疫力之 功效。 根據本發明之促進神經膠瘤細胞產生活性氧化物之方 98.12.10第96130689號專利說明書及申請專利範圍修正本 法4其包3步驟：提供—促進液，且該促進液至少伐 曰序驗之成分’及將該促進祕予—神經_細胞.2 ’該神_瘤細胞係受外界微生物侵人。 、中 【實施方式】 “為讓本翻之上述及其他目的、特徵、優點 易幢’下文特舉本發明之較佳實施例，並配 頭 作詳細說明如下： 圖式， 。月參知第1圖所示，本發明較佳實施例之促 瘤細胞產生活性氧化物之方法的第一步驟81係：提供申7 進，丄且該促進液至少係包含香芹酚（carvacrol)之成分足 更詳言之’該促進液係用以促進一神經膠瘤細胞 氧化物，該香“之分子式係為CigH]4〇,且該料^ 為一液體，該香芽盼不可溶於甘油，但可溶於水或乙醇^ ethyl alcohol) ’目此該促進液内可另包含—轉劑，以盘 混合並調整該編之濃度，且該稀釋劑係可選 擇為水或乙醇。 、 明再參知、第丨圖所示，本發明之第二步驟係. =液施予-神鄉瘤細胞。更詳言之，將該促進液施= 神經膝瘤細胞，以使該神經膠瘤細胞可吸收該促進液内^ 香芽紛成分，而促進該神經膠瘤細胞產生活性氧化物 將該神經勝瘤細胞於一環境中進行培養，並加入該促進 ’以增進該神經滕瘤細胞之活性氧化物之產生量。 用以：：，如前述，該神經膠瘤細胞内之活性氧化物雖可 寸病原體，然而，若該神經膠瘤細胞内之活性氧化 7 I32146998· 1Ζ· 1〇第96讓89號專利說明書及申請專利範圍修正本 物過量時，該雜魏物會轉職、蛋㈣ ’反而將造成不良影響’因此，若該神經膠瘤細胞並未受^ 外界微生物入侵，則該活性氧化物維持適量即可；反之，若外 界微生物侵入，則較佳係適當觸發該活性氧化物產生，進而促 進免疫功能。本發明之促進細胞產生活性氧化物之方法，於 該神經勝瘤細胞未受體外微生物入侵時，則該神經廢瘤細 胞將不會受到該促進液之觸發而增加活性氧化物之生成量 ，因此，不會因過量之活性氧化物而受損；反之，於外界 微生物入侵該神經膠瘤細胞時，則該促進液將觸發該神經 膠瘤細胞產生較多量活性氧化物，以f助殺除該微生物， 進而增進免疫力。 為驗證本發明之钱細胞產生活性氧化物之方法確 實具有確實具有提升制裕度、活性氧化物產生量及細胞免 疫力之功效’因此另進行細胞存活率分析及活性氧化物產 生量分析，以確認本發明之細胞毒性及活性氧化物之產生 量0 请參照第2圖所示，本發明之細胞存活率分析係以 MTT assay進行力析，以確認該促進液内所含之香芹齡對 該神經膠瘤細胞是否具有生物毒性，於下列試驗中，該促 進液係選擇另包含水以調整該香芹酚之濃度。該神經膠瘤 細胞存活率分析之實施步驟係：首先，選擇將〇2mg之 MTT〔 3-(4,5-dimethylthiaz〇l-2-yl)-2,5-diPhenyl tetrazolium bromide〕（sigma)加入imi培養液中，以配製成濃度為 0.2mg/ml之MTT溶液，且該MTT溶液較佳係避光保存， 98.12.10 第96130689號專利說明 書及申請專利範圍修正本 以免變質’接著’計數—細胞以配製絲亳升具有㈣5 個細胞之細胞液，並選擇取之細胞液於一 培養皿中料】2小時後，更_的RPM料液，並加入 該促進液，且該促進㈣之香㈣（sigma)濃度係選擇為 〇至·/ZM。置放於3rc且含有5%二氧化碳（c〇2)之 培養I目内培養24小時後，以吸取方式去除培養液，接著以 每酉夂鹽緩衝液（phosphate buffer saline, PBS )洗條細胞後 ’加入濃度為2mg/ml的MTT溶液50//L於各該培養皿中 ’且較佳係避光保存’接著’置於37。(：且含有5%二氧化 石厌之培養箱中培養3小時’至此，存活之細胞内的琥珀酸 去氫酶將把MTT轉換成藍紫色結晶，接著，移除MTT後 加入 50//L 之二曱基亞楓（dimethyl sulfoxide, DMSO)，以 將該神經膠瘤細胞中之藍紫色結晶溶出，最後以ELISA reader (Bio-TEK)在570nm之波長下測其吸光值，以控制 組的吸光值當作100%存活率，而實驗組（加入含適當濃度 香芹酚之促進液）和控制組（未加入促進液）之比值則為 相對之細胞存活率。其中，該神經膠瘤細胞係為glioma C6 (食品工業研究所），且該鱗酸鹽缓衝液係以0.¾之氯化 鉀（KC1)、〇.24g之磷酸二氫鉀（KH2P〇4)、8§氯化鈉（ NaCl)及144g之磷酸氫二鈉（Na2HP〇4)共同混合溶解 於1L之去離子水（2d由0)中，且pH值係為7.2至7.4 請再參照第2圖所示，其係為對該促進液内所含之香 芹驗進行細胞存活率分析後之結果，由結果可仔知’加入 1321469 98. 12.10第96130689號專利說明書及申請專利範圍修正本 含〇〜500香芽齡之促進液後，與控制組（c)相比，並 沒有達到50。/。細胞死亡，因此判斷在香芹酚在5〇〇#M以下之 濃度對細胞沒有毒殺性。亦即，本發明之促進細胞產生活性 氧化物之方法對細胞確實不具有毒性。… 請參照第3及4圖所示’該活性氧化物產生量分析之 步驟係：將每毫升含有5χ105個細胞（神經膠瘤細胞）之 細胞液取lml種於12well培養皿中，於37。〇且含有5% _ 氧化碳之培養箱中培養至該神經膠瘤細胞貼附，接著選擇 加入濃度為 1 /z g/ml 之脂多酶（lip〇p〇iySaccharide, LPS )、 400ng/ml之TPA或含100至200从]\4香芹紛之促進液共同 作用’該脂多醣係為細菌萃取物，以模擬外界微生物，該 TPA係用以增加對該脂多醣之刺激，接著，於作用3〇分鐘 後加入濃度為20M之二氯螢光素二乙酸酯（ 2?,7,-dichlorodihydrofluorescein diacetate, DCFH-DA )( calbiochem)選擇作用30分鐘，以藉由該二氯螢光素二乙 酸酯與該神經膠瘤細胞所產生之活性氧化物結合並發出鸯 光’接著，將該RPMI培養液去除，並以該磷酸鹽緩衝液 清洗一次，再將該碟酸鹽缓衝液去除，接著，加入 之胰蛋白酵素（Trypsin-EDTA，Gibco)，以將該神經膠瘤細 胞從培養皿上拍下，再選擇加入lml之磷酸鹽緩衝液打散 細胞，最後以細胞流速儀（flow cytometry, Becton Dickinson ;FACScan, California)分析該神經膠瘤細胞内自由基的產 生量。 請參照第3圖所示，其係為該活性氧化物產生量分析 —10 — 1321469 • 98.12·10第9613_號專利說明書及申請專利範圍修正本 之螢光表現量相對細胞數之變化圖。复中，第 制組’亦即該細胞液中並未添加酿 促=控 .之、促進液共同作用，其中，該腊多醣 =度1心如，該TPA之濃度麵伽 =則敛述於活性氧化物產生量分析之步驟中，於此;： 组相第3圖麻，由結料得知，^組與第C 促、隹^合可知H細胞液中加入含2 0 0 " Μ香芽齡之 a =:::r=:r生活性氧化物. 二，環境下將產生活性氧；:==== d、，且可發現，於脂乡較TpA存在 …'弟 :=進該神二二： ::生物入侵…將不會觸發該神經膠 :卜 ，，以避免對該神經膠瘤細胞造 = _瘤細胞受外界微生物入侵之環境下，二4= 促進液將有效觸發該神經膠瘤細胞產生較多量之 物，以幫助殺除該外界微生物，進而提 /虱化 ;發明確實具有提升活性氧化物產生量及細:免疫::功 98.12.1D帛％讓89號專利說明書及中請專利範圍修正本 請參照f 4 1料’其齡不同香㈣濃度之促進液 下3活ϋ氧化物產生量分析之榮光表現量相對細胞數之 义化圖’、中第6組係為控制组，亦即該細胞液中並未 添加脂多醣、ΤΡΑ或促進液；第仏之、細胞液中係添加脂 多醣及TPA共同作用，並未添加該促進液；第g組之細胞 液中係添加脂多酿、取及含100 W之香芽紛之促進液 /、同作用’第h組之細胞液中係添加脂多_、TPA及15〇 之香㈣共同作用；第i、组之細胞液中係添加脂多酶 ，曲/及GG//M之香芽紛共同作用，其中，該脂多釀之 派度係為lyg/ml，該TPA之濃度係為她娜，其他條 件則敘述於活性氧化物產生量分析之步驟中，於此不再資 述。 Μ料照弟4 _示，由結果可得知，隨著該香料 派又之增加，該活性氧化物之產生量亦有上升之趨勢。麥 此可侍知本發明之方法確實具有提升活性 細胞免疫力之功效。 里及 ▲如上所述’相較於f用殺菌劑僅適用於由密螺旋體屬 所=成之麵，進*造錢驗度減之缺點；再者，該習 殺菌劑僅係m組成物，並未提供-促進細胞產生活性氧 H之方法’使付生物體内的活性氡化物產生量可能較為低落 。械，本發明係將—至少含有香料之促紐施予神經膠 瘤、”田胞’使該神經顯細胞於外界微生物人侵時，可 ，相膠瘤細胞_性氧化物之產生量。藉此，本發明確 貫可提升使用裕度、活性氧化物產生量及細胞免疫力。 —12 — 1321469 98. 12· 10第96130689號專利說明書及申請專利範圍修正本 雖然本發明已利用上述較佳實施例揭示，然其並非用 以限定本發明，任何熟習此技藝者在不·本發明之精神 和範圍之内，相對上述實施例撕各種更動與修改仍屬本 發明所保護之技術範《#，因此本發明之倾 後附 之申請專利範圍所界定者為準。 —13 ~ 1321469 98. 12. 10第96130689號專利說明書及申請專利範圍修正本 ί 【圖式簡單說明】 第1圖：本發明之促進神經膠瘤細胞產生活性氧化物之 方法的流程方塊圖。 第2圖：本發明之細胞存活率分析結果。 第3圖：本發明之活性氧化物產生量分析之螢光表現量 相對細胞數之變化圖。 第4圖：本發明於含有不同濃度香芹酚之促進液下，活 性氧化物產生量分析之螢光表現量相對細胞數之變化圖。 【主要元件符號說明】 S1 第一步驟 S2 第二步驟 —14 —98.12.10 Patent Specification No. 96130689 and the scope of the patent application. The present invention relates to a method for promoting the production of active oxides in glioma cells, in particular, at least The propellant (carvacr〇1) promoting solution is administered to a glioma cell, so that the glioma cell can increase the amount of active oxide produced by external microbial invasion. 'Use margin and cellular immunity promote cell production. A method of active oxides. [Prior Art] Generally, cells in an organism are susceptible to the microorganisms of the organism due to the invasion of various microorganisms (the micha rganism), thereby causing damage to the cells of the organism and causing the organism to cause damage to the organism. Due to its anti-S and g-killing effects, the body has been used to kill only bacteria and microorganisms. For example, the conventional fungicides, such as the Republic of China Patent No. 28975, "Use as a fungicide The fragrant and thyme tests showed that they contained fragrant (4) and musk turn. The fragrant two tongues were 5P to 9〇% dry weight, and the thymol was 5ppm to 80g. 'And the weight ratio of the fragrance to the thyme is 1: 5 to i〇: i is used in diet composition, drinking supplement or bath supplement to treat the genus Treponema The disease caused, such as _ illness, etc.), is the active oxide of reactive material (reactive (10) cis 1 sPecies, ROS, the molecule of the oxidative force of the directional oxidative force] when the active oxide in the cell is excessive The time of the society, the Wei Wei will attack the ugly, the egg is self-quality and the fun (4) In turn, it causes ',, repairs (four) wealth, fine, riding oxides are also necessary for life 1321469 98. 12. 10 Patent No. 96130689 and the scope of the patent application, such as superoxide anion ((4) Bottle ide anion), it is white blood glass used to pay for the disease «shirt, therefore, if the raw _ and raw === oxide silk hold her; vice versa, if 峨 = ^ into the organism, then the body The preferred system suitably triggers the production of the active oxide, thereby promoting the immune function in the living body. As a general matter, the above-mentioned conventional bactericide has the following disadvantages, for example, the bactericidal sputum is caused by the dense spiral, Further, the use of the margin storage slit; in addition, the (four), does not provide - strip = = = the amount of active oxide produced in the body may be relatively low. For the above reasons, it is necessary to further improve the above-mentioned conventional fungicide. In view of the above, the present invention improves the above-mentioned disadvantages by administering at least t-promoting promoting fluid to a glioma cell, so that the glioma cell can be raised by external micro-invasion Endothelium reactive oxygen species The production amount of the compound 'by this' can indeed improve the use margin, the amount of active oxide production and the cellular immunity. SUMMARY OF THE INVENTION The main object of the present invention is to provide a method for promoting the production of active oxides in glioma cells. The invention can increase the amount of active oxides produced in the glioma cells by administering at least the stimulating solution containing the fragrant buds to the glioma cells, so that the glioma cells can be invaded by external microorganisms, so that the present invention The utility model has the advantages of improving the use margin, the amount of active oxide production and the cellular immunity. The method for promoting the production of active oxides of the glioma cells according to the invention 98.12.10 Patent No. 96130689 and the scope of the patent application are amended. Package 3: Providing a promoting liquid, and the promoting liquid is at least a component of the test, and the promoting secret is - the nerve cell. 2 'The god cell line is invaded by external microorganisms. [Embodiment] "In order to make the above and other objects, features, advantages and advantages of the present invention", the preferred embodiment of the present invention will be described below, and the heading will be described in detail as follows: Fig., 1 is a first step 81 of a method for producing an active oxide by a tumor-promoting cell according to a preferred embodiment of the present invention, wherein the promoting solution contains at least a component of carvacrol. More specifically, the promoting liquid system is used to promote a neuroglioma cell oxide, the molecular formula of which is CigH]4〇, and the material is a liquid, and the fragrance is insoluble in glycerin, but Soluble in water or ethanol ^ ethyl alcohol) 'The purpose of this promoting liquid may additionally include - transfer agent, the disc is mixed and adjusted the concentration of the braid, and the diluent may be selected from water or ethanol. As shown in the figure, the second step of the present invention is: liquid administration-shenzhen tumor cells. More specifically, the promoting fluid is applied to the neurokaryngeal tumor cells, so that the neuroglioma cells can absorb the components of the promoting liquid, and promote the production of active oxides of the neuroglioma cells to win the nerves. The tumor cells are cultured in an environment, and the promotion is added to increase the amount of active oxide produced by the neuron cell. For::, as mentioned above, the active oxide in the glioma cell can be pathogenic, however, if the active oxidative activity in the glioma cell is 7 I32146998·1Ζ·1〇, 96, the patent specification No. 89 and When the scope of application for patents is corrected, the amount of the Weiwei will be transferred, and the egg (4) will adversely affect. Therefore, if the glioma cell is not invaded by external microorganisms, the active oxide can be maintained in an appropriate amount. On the other hand, if external microorganisms invade, it is preferable to appropriately trigger the production of the active oxide, thereby promoting the immune function. The method for promoting the production of active oxides by the cells of the present invention, when the nerve cells of the nerves are not invaded by the microorganisms outside the receptor, the nerve tumor cells will not be triggered by the promoting liquid to increase the amount of active oxides. Therefore, it will not be damaged by excessive active oxides; conversely, when the external microorganism invades the neurotrophic cells, the promoting liquid will trigger the production of a larger amount of active oxides by the glioma cells, so as to help kill The microorganism enhances immunity. In order to verify that the method for producing active oxides of the money cells of the present invention does have the effect of improving the production margin, the amount of active oxide production, and the cellular immunity, the cell survival rate analysis and the activity of the active oxide production are further analyzed. The amount of cytotoxicity and active oxide produced by the present invention was confirmed. Referring to Fig. 2, the cell viability analysis of the present invention was carried out by MTT assay to confirm the age of the celery contained in the promoting solution. Whether the glioma cells are biologically toxic, in the following experiments, the promoting fluid system additionally comprises water to adjust the concentration of the carvacrol. The steps of the analysis of the viability of the neuroglioma cells are as follows: First, the addition of 2 mg of MTT [ 3-(4,5-dimethylthiaz〇l-2-yl)-2,5-diPhenyl tetrazolium bromide] (sigma) is added. In the imi culture solution, the MTT solution is prepared at a concentration of 0.2 mg/ml, and the MTT solution is preferably stored in the dark, 98.12.10 Patent No. 96130689 and the scope of the patent application are modified to avoid deterioration. - the cells are prepared to have a cell fluid of (4) 5 cells, and the cell liquid is selected to be in a petri dish. After 2 hours, the RPM solution is further added, and the promoting solution is added, and the promoting (4) is The concentration of sigma was chosen to be 〇 to / / ZM. After culturing in culture medium I containing 3 cc of carbon dioxide (c〇2) for 24 hours, the culture solution was removed by suction, followed by washing the cells with phosphate buffer saline (PBS). 'Add 50/L of MTT solution at a concentration of 2 mg/ml in each of the culture dishes 'and preferably protected from light' followed by 'at 37'. (: and cultured for 3 hours in an incubator containing 5% dioxide.) The succinate dehydrogenase in the surviving cells will convert the MTT to blue-violet crystals, then remove the MTT and add 50//L. Dimethyl sulfoxide (DMSO) to dissolve the blue-violet crystals in the glioma cells, and finally absorb the absorbance at 570 nm using an ELISA reader (Bio-TEK) to control the group The absorbance value was taken as 100% survival rate, and the ratio of the experimental group (adding the promotion liquid containing the appropriate concentration of carvacrol) to the control group (not adding the promotion liquid) was the relative cell survival rate. The system is glioma C6 (Institute of Food Industry), and the sulphate buffer is 0.3⁄4 potassium chloride (KC1), 2424g potassium dihydrogen phosphate (KH2P〇4), 8§ sodium chloride. (Nas) and 144g of disodium hydrogen phosphate (Na2HP〇4) are mixed and dissolved in 1L of deionized water (2d by 0), and the pH value is 7.2 to 7.4. Please refer to Figure 2 again. In order to analyze the cell viability of the parsley contained in the promoting solution, it is known from the results that 'Add 13 21469 98. 12.10 Patent Specification No. 96130689 and the scope of the patent application. After the promotion solution containing 〇~500 fragrant buds, compared with the control group (c), it did not reach 50%. The concentration of celery at 5 〇〇 #M or less is not toxic to cells. That is, the method for promoting the production of active oxides by cells of the present invention is not toxic to cells.... Please refer to Figures 3 and 4 The step of analyzing the amount of active oxide production is as follows: 1 ml of cell liquid containing 5 χ 105 cells (neurocolloma cells) per ml is seeded in a 12well culture dish in 37. 〇 and containing 5% _ carbon oxide in an incubator Incubate to the glioma cell for attachment, followed by selective addition of lipopolyzyme (lipsp〇iySaccharide, LPS) at a concentration of 1 /zg/ml, TPA at 400 ng/ml or 100 to 200 from the carcass The synergistic effect of the promoting liquid 'the lipopolysaccharide is a bacterial extract to simulate the external microorganisms. The TPA is used to increase the stimulation of the lipopolysaccharide. Then, after 3 minutes, the concentration of 20M dichlorofluoride is added. Photodiacetate (2?,7,-dichlorodi Hydrofluorescein diacetate, DCFH-DA ) ( calbiochem ) is selected for 30 minutes to bind to the active oxide produced by the neurotrophic cell by the dichlorofluorescein diacetate and to emit a calender. The RPMI medium was removed and washed once with the phosphate buffer, and the discate buffer was removed. Then, trypsin (EDpsin-EDTA, Gibco) was added to remove the neuroadoma cells from the Petri dish. The above photograph was taken, and then 1 ml of phosphate buffer was added to break up the cells. Finally, the amount of free radicals in the neuroglioma cells was analyzed by flow cytometry (Becton Dickinson; FACScan, California). Please refer to Fig. 3, which is a graph showing the change of the amount of fluorescence expression relative to the number of cells in the analysis of the active oxide production amount - 10 - 1321469 • 98.12·10, No. 9613_ and the patent application scope revision. In the middle of the group, the first group 'is not added to the cell liquid, and the promoting liquid has a synergistic action. The waxy polysaccharide=degree 1 heart, and the concentration of the TPA is condensed. In the step of analyzing the amount of active oxide production, here: the third phase of the phase is obtained, and it is known from the nodules that the group of groups and the C group promotes that the H cell liquid is added with the content of 2 0 0 " A germination age a =:::r=: r living oxide. Second, the environment will produce active oxygen;: ==== d, and can be found in the fat town than TpA exists ... 'di: = Into the god 22: :: biological invasion ... will not trigger the nerve gel: Bu, to avoid the environment of the neuroglioma cells = _ tumor cells invaded by external microbes, the second 4 = promoting fluid will be effective Triggering the glioma cells to produce a larger amount of substances to help kill the external microorganisms, and then extracting/deuterating; the invention does increase the amount of active oxides produced and fine: immunization:: work 98.12.1D帛% let the number 89 For the patent specification and the revision of the patent scope, please refer to the glory of the analysis of the amount of 3 active bismuth oxides produced under the promotion of the concentration of different fragrances (4). The current amount of relative cell number is 'the map', the middle group is the control group, that is, the cell liquid is not added with lipopolysaccharide, sputum or promoting solution; the second, cell liquid is added with lipopolysaccharide and TPA In combination, the promoting solution was not added; in the cell liquid of the g group, the fat was added, and the boosting liquid containing 100 W of the fragrant buds was added, and the cell liquid in the same group was added. More _, TPA and 15 〇 香 (4) work together; the i, group of cell fluid is added with lipopolyzyme, 曲 / and GG / / M buds have a joint effect, which, the fat more brewing The system is lyg/ml, the concentration of the TPA is Herna, and other conditions are described in the step of analyzing the amount of active oxide production, which is not described here. According to the results, it can be seen from the results that as the fragrances increase, the amount of active oxides also increases. This allows the method of the present invention to have the effect of enhancing the immunity of the active cells. In the above and ▲ as described above, compared with the fungicide used in f, it is only applicable to the surface of the genus of the genus Tremella, which is the disadvantage of reducing the amount of money; in addition, the bactericide is only composed of m composition, There is no provision - a method to promote the production of reactive oxygen species by cells - to make the amount of active telluride produced in the living organisms relatively low. The invention relates to a method in which at least a spice is added to a neuroglioma, and a "cell" is used to cause the nerve cells to invade by external microorganisms. Thus, the present invention can improve the use margin, the amount of active oxide production, and the cellular immunity. 12- 1321469 98. 12·10 No. 96130689 Patent Specification and Patent Application Revision Although the present invention has utilized the above preferred The embodiments are not intended to limit the present invention, and those skilled in the art are not able to omise various modifications and modifications to the above-described embodiments within the spirit and scope of the present invention. Therefore, the scope of the patent application of the present invention is defined by the scope of the patent application. -13 ~ 1321469 98. 12. 10 Patent No. 96130689 and the scope of the patent application amendments 【 [Simple description of the drawings] Figure 1: This A block diagram of a method for promoting the production of active oxides by neuroglioma cells of the invention. Fig. 2: Results of cell viability analysis of the present invention. Fig. 3: Production of active oxides of the present invention A graph showing the change in the amount of fluorescence expression relative to the number of cells in the amount of analysis. Fig. 4 is a graph showing the change in the amount of fluorescent expression versus the number of cells in the amount of active oxide produced by the present invention in a solution containing different concentrations of carvacrol. [Main component symbol description] S1 First step S2 Second step - 14 —