200908954 九、發明說明： 【發明所屬之技術領域】 本發明係關於一種促進細胞產生活性氧化物之方法 ，特別疋關於將至少含有香序g分（carvacr〇l)之促進液施 予一細胞，使該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】 一般而言，一生物體中之細胞由於外界各種微生物（ m1Cr〇organism)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 變。 ^ 香芹酚由於具有抗菌及殺菌效果，因此已應用於殺除特 疋之病菌及微生物，舉例而言’ „f用殺制，如中華民國專 利第1228975號「使用作為殺菌劑之含香序酴及靡香草酉分之 組成物」發明專觸示，其係包含香絲及麝香㈣，該香 序酚成分縣5PPm ^ 9〇%乾冑，_香草義為5聊至8〇 %乾重，且該香㈣對麝香轉之重量比係為丨：5至ι〇 :工 ，其係應祕飲食組祕、飲水補充物或浸浴補充物中，以治 療由密螺频屬（Trep。職a)所造狀絲，如義疾等。 另’生物體内之活性氧化物（reactlve〇xygenspe⑽，R〇s )，為具有高魏化力的分子，#細翻騎性氧化物過量時 ’、，活性氧化齡攻擊職、蛋的及細胞_f，進而造 成热法修復的傷害，然而，該活性氧化物對生命而言亦是必須 PK10417 2007/8/20 200908954 以針什二ί ΐ (SUper〇Xlde ani〇n) ’則是白血球用 以對付病原體之武器，因此，若生物體内並未受到外界微 入侵，則騎性氧化物係維持適4即可；反之，若外界微 侵入該生物體内，則體内較佳係適當觸發該活 ： 進而促進該生物體内之免疫功能。 物產生， 一般而言’上述習用殺菌劑具有下列缺，點，例如”玄 習用殺菌雜適用於由密螺旋體屬所造成之疾病，進而造= 使用裕度低落之缺點；再者，該習用殺菌劑僅係為—殺菌^成 物，並未提供-促進細胞產生活性氧化物之方法，使得节生物 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。 121 練於此，本發良上述之缺點，其係經由將至少 含有香芽S分之促進液施予-細胞，使該細胞於外界微生物 入侵時，可提升該細胞内活性氧化物之產生量，藉此，本 發明確實可提升使轉度、活性氧化物產生量及㈣免疫力 〇 【發明内容】 本發明主要目的储供—種促進細胞產生活性氧化 物之方法’其係經由將至少含有香序酴之促進液施予一細 胞，使該細胞於外界微生物入侵時，可提升該細胞内活性 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。 根據本發明之促進細胞，其包含步驟：提供一促進液 ’且該促進液至少係包含香芽紛之成分；及藉由一施劑方 PK10417 2007/8/20 200908954 法將D亥促進液施予一生物體之細胞；其中，該生物體之細 胞係受外界微生物侵入。 【實施方式】 ^為讓本發明之上述及其他目的、特徵、優點能更明顯 易丨董下文特舉本發明之較佳實施例，並配合所附圖式， 作詳細說明如下： 5月茶照第1圖所示，本發明較佳實施例之促進細胞產 生活性氧化物之方法的第一步驟S1係：提供一促進液，且 。亥促進液至少係包含香芹紛（)之成分。更詳言之 ，忒促進液係用以促進一細胞產生活性氧化物，該香芹酚 之分子式係為C^HhO，且該香芹酚係為一液體，該香芹 酚不可溶於甘油，但可溶於水或乙醇（ethyl alc〇h〇1)，因 此该促進液内可另包含—稀釋劑，以與該香㈣混合並調 整該香;^之濃度，且該稀釋劑係可選擇為水或乙醇。 凊再茶照第1圖所示，本發明之第二步驟係：藉由一 施劑方法將該促進液施予一生物體之細胞。更詳言之，將 該促進液針該生物體之細胞，喊該細胞可吸收該促進 液内之香？f喊分’該生物體係可選擇為人體或其他生物 體。該施劑方法係可選擇以食人、飲人m及入等方 式將該促進液送人該生物體_，並㈣生減内被吸收 而促進該細胞產生活性氧化物；亦可選擇於該生物體體外 以浸泡或塗抹於該生物財面之方式，使祕進液直融 該生物體之細胞_而增進活性氧化物之生成量； 可將細胞直涵該生物體分離，料―環境中進行ς養，、 ΡΚ10417 2007/8/20 200908954 並加 人父 述，該細胞内之活性氧化物雖可用以二付 入以生物體内之病，然而，^勿^用以對付 物過量時，該活性氧化物會攻擊DNA、蛋二，活性乳化 ，反而將賴生無造成不㈣響，因此蛋胞膜脂質 ❹__人侵，_性氧化= 場細，職難姆輸活性 虱匕物產生，進而促進該生物體内之免疫功能。本發明之促進 細胞產生活性氧化物之方法，於該生物體未受體外微生物 =侵時，則該細胞將不會受到該促進液之觸發而增加活性 乳化物之生成量’因此，該生物體不會因過量之活性氧化 物而受損；反之’該生物體於外界微生物人侵時，則該^ 進液將觸發該生物體之細胞產生較多量活性氧化物，以幫 助δ亥生物體殺除該微生物，進而增進該生物體之免疫力。 為驗證本發明之促進細胞產生活性氧化物之方法確 貫具有確實具有提升使用裕度、活性氧化物產生量及細胞免 疫力之功效，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。 請參照第2圖所示，本發明之細胞存活率分析係以 MTT assay進行分析，以確認該促進液内所含之香芽g分對 該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。該細胞存活率分析之 實施步驟係：首先，選擇將〇.2mg之MTT〔 PK10417 2007/8/20 200908954 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide〕（sigma)加入lml培養液中，以配製成濃度為 0.2mg/ml之MTT溶液’且該Μττ溶液較佳係避光保存， 以免變質，接著，計數一細胞以配製成每毫升具有1χ1〇5 個細胞之細胞液，並選擇取l〇〇yL之細胞液於一 96well 培養皿中培養12小時後，更換新的RPMI培養液，並加入 該促進液，且該促進液内之香芹酚（Sigma)濃度係選擇為 0至500//Μ。置放於37°C且含有5%二氧化碳（c〇2)之 培養箱内培養24小時後，以吸取方式去除培養液，接著以 磷酸鹽緩衝液（phosphate buffer saline，PBS)洗滌細胞後 ，加入濃度為2mg/ml的MTT溶液50//L·於各該培養孤中 ，且較佳係避光保存，接著，置於37t：且含有5%二氧化 一|]碳之培養箱中培養3小時，至此，存活之細胞内的玻珀酸 去氫酶將把MTT轉換成藍紫色結晶，接著，移除後 加入 50 "L 之二甲基亞楓（dimethyl sulfoxide, DMSO)，以 將該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，該細胞係選擇為神經膠瘤細胞（gli〇ma C6，食品工業研究所）’且該磷酸鹽緩衝液係以〇 2g之氣 化鉀（KC1)、〇.24g之磷酸二氫鉀（Kh2P〇4)、8g氣化銅 (NaCl)及l.44g之磷酸氫二鈉（Na2HP〇4)共同混合溶 解於1L之去離子水（2d氏0)中，且PH值係為7.2至7.4 PK10417 2007/8/20 200908954 請再參照第2圖所示，其係為對該促進液内所含之香 芹酚進行細胞存活率分析後之結果，由結果可得知’加入 含〇〜500#M香芹酚之促進液後，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 /辰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5xl05個細胞（神經膠瘤細胞）之 細胞液取lml種於I2well培養ιοί中，於37°C且含有5%二 氧化碳之培養箱中培養至該細胞貼附，接著選擇加入濃度 為 1 // g/ml 之脂多醣（lip〇p〇lysaccharide, LPS )、400ng/ml ]|]之ΤΡΛ或含100至200//M香芹酚之促進液共同作用，該 脂多醣係為細菌萃取物，以模擬外界微生物，該ΤΡΑ係用 以增加對該脂多醣之刺激，接著，於作用30分鐘後加入濃 度為 20Μ 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 酸酯與該細胞所產生之活性氧化物結合並發出螢光，接著 ，將該RPMI培養液去除，並以該填酸鹽緩衝液清洗一次 ，再將該磷酸鹽緩衝液去除，接著，加入100//1之胰蛋白 酵素（Trypsin-EDTA，Gibco)，以將該細胞從培養皿上拍下 ，再選擇加入lml之礙酸鹽緩衝液打散細胞，最後以細胞 流速儀（fl〇w cytometry, Becton Dickinson ; FACScan， PK10417 2007/8/20 —10 — 200908954200908954 IX. Description of the Invention: [Technical Field] The present invention relates to a method for promoting the production of an active oxide by a cell, and more particularly to administering a promoting solution containing at least a carotropin to a cell. When the cells are invaded by external microorganisms, the amount of active oxides generated, the use margin, and the cellular immunity are promoted to promote the production of active oxides by the cells. [Prior Art] In general, cells in an organism are susceptible to the influence of various microorganisms (m1Cr〇organism), and the cells of the organism are susceptible to the microorganisms, thereby causing damage to the cells of the organism and causing the organism to cause damage to the organism. The body produces lesions. ^ Carvacrol has been used as an antibacterial and bactericidal effect, so it has been used to kill the pathogens and microorganisms of the cockroaches. For example, ‘f kills, such as the Republic of China Patent No. 1228975, “Uses as a fungicide The composition of 酴 and 靡 酉 」 」 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 四 四 四 四 四 四And the weight ratio of the fragrance (4) to musk is 丨: 5 to ι〇: work, which is secreted by the diet group, drinking water supplement or bath supplement, to treat by Trep. Job a) made of silk, such as Yi disease. In addition, the active oxide in the living body (reactlve〇xygenspe(10), R〇s) is a molecule with high Weihua force, #fine ride-on oxide when excess, 'active oxidized age attacking, egg and cell_f , in turn, causing damage to the thermal repair, however, the active oxide is also necessary for life PK10417 2007/8/20 200908954 to the needle (Uper〇Xlde ani〇n) 'is the white blood cell to deal with The weapon of the pathogen, therefore, if the organism is not subject to external micro-invasion, the riding oxide system can be maintained at 4; on the contrary, if the outside micro-invasion into the living body, the body is better to trigger the activity. : Further promoting the immune function in the living body. In general, the above-mentioned conventional fungicides have the following disadvantages, such as, for example, "the use of bactericidal bacteria for the disease caused by the genus Treponema, and then the disadvantage of using a low margin; and further, the conventional sterilization The agent is only a bactericidal compound, and does not provide a method for promoting the production of active oxides by the cells, so that the amount of active oxides produced in the knuckles may be relatively low. For the above reasons, it is necessary to further improve the above-mentioned conventional bactericides. 121 In this case, the above-mentioned disadvantages of the present invention are achieved by administering a cell containing at least a scented scented S-cell to enhance the production of active oxides in the cell when the cell is invaded by external microorganisms. Therefore, the present invention can improve the degree of rotation, the amount of active oxide generated, and (4) the immunity. [The present invention] The main purpose of the present invention is to provide a method for promoting the production of active oxides by cells. The promoting liquid containing the scented scorpion is administered to a cell, so that when the cell invades by the outside microorganism, the amount of active oxide in the cell can be increased, so that The present invention has the effects of improving the use margin, the amount of active oxide production, and the cellular immunity. The promoting cell according to the present invention comprises the steps of: providing a promoting liquid and the promoting liquid comprises at least a component of the fragrant bud; And applying a D-Promoting solution to a cell of an organism by a method of applying PK10417 2007/8/20 200908954; wherein the cell line of the organism is invaded by an external microorganism. [Embodiment] ^ For the purpose of the present invention The above and other objects, features, and advantages will be more apparent from the following description of the preferred embodiments of the invention. The first step S1 of the method for promoting the production of active oxide by the cells of the preferred embodiment is: providing a promoting liquid, and the at least promoting liquid comprises at least the component of the celery (). More specifically, the sputum promoting liquid system is used. In order to promote the production of an active oxide by a cell, the formula of the carvacrol is C^HhO, and the carvacrol is a liquid, the carvacrol is insoluble in glycerin, but soluble in water or ethanol (ethyl alc) 〇h〇1), Therefore, the promoting liquid may further comprise a diluent to mix with the fragrance (4) and adjust the concentration of the fragrance; and the diluent may be selected from water or ethanol. The second step of the invention is: applying the promoting solution to the cells of a living body by an application method. More specifically, the promoting liquid is needled to the cells of the living body, and the cells are called to absorb the promoting liquid. Xiang?f shouting 'The biological system can be selected as the human body or other organisms. The application method can choose to send the promoting liquid to the living body by means of eating, drinking, and ingesting _, and (4) birth reduction The inside is absorbed to promote the production of active oxides of the cells; or the body may be selected to be immersed or smeared on the biological surface in a manner to allow the secret solution to directly melt the cells of the organism. The amount of production; the cells can be directly culminated in the organism, and the material is maintained in the environment, ΡΚ10417 2007/8/20 200908954 and added by the father, the active oxides in the cells can be used to pay in two The disease in the body, however, ^Do not use it to deal with things When the amount, the active oxide will attack DNA, egg two, active emulsification, but will not cause no (four) ring, so the egg membrane lipid ❹ __ human invasion, _ sexual oxidation = field fine, job hard to lose activity 虱The sputum is produced to promote immune function in the living body. The method for promoting the production of an active oxide by a cell of the present invention, wherein the organism does not receive the triggering of the promoting solution to increase the amount of active emulsifier generated when the microorganism is not exposed to the receptor. The body will not be damaged by excessive active oxides; otherwise, when the organism invades the external microorganisms, the liquid will trigger the cells of the organism to produce a larger amount of active oxides to help the organisms The microorganism is killed, thereby enhancing the immunity of the organism. In order to verify that the method for promoting active cell production of the present invention has the effect of improving the use margin, the amount of active oxide production, and the cellular immunity, the cell survival rate analysis and the amount of active oxide production are analyzed. The cytotoxicity and the amount of active oxide produced by the present invention were confirmed. Referring to Fig. 2, the cell viability analysis of the present invention was analyzed by MTT assay to confirm whether the aroma g-containing fraction contained in the promoting solution is biologically toxic to the cell, and the promotion was carried out in the following tests. The liquid system selection additionally includes water to adjust the concentration of the carvacrol. The cell survival rate analysis is carried out by first selecting: MTT [PK10417 2007/8/20 200908954 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] Sigma) is added to 1 ml of the culture solution to prepare a MTT solution having a concentration of 0.2 mg/ml' and the Μττ solution is preferably stored in the dark to avoid deterioration. Then, one cell is counted to have a ratio of 1 每 per ml. 〇 5 cells of the cytosol, and select the cell solution of l〇〇yL for 12 hours in a 96well culture dish, replace the new RPMI culture solution, add the promotion solution, and the parsley in the promotion solution The concentration of phenol (Sigma) was selected to be 0 to 500 / / Μ. After being cultured in an incubator containing 5% carbon dioxide (c〇2) at 37 ° C for 24 hours, the culture solution was removed by suction, and then the cells were washed with phosphate buffer saline (PBS) and then added. The MTT solution having a concentration of 2 mg/ml is 50//L in each of the culture cells, and is preferably stored in the dark, and then placed in an incubator of 37 t: and containing 5% dihydrate|carbon] Hours, at this point, the peroxidic dehydrogenase in the surviving cells will convert the MTT to blue-violet crystals, then remove and add 50 "L dimethyl sulfoxide (DMSO) to The blue-violet crystals in the cells were eluted, and finally the absorbance was measured by ELISA reader (Bio-TEK) at a wavelength of 570 nm. The absorbance of the control group was taken as the 100% survival rate, and the experimental group (added with the appropriate concentration of parsley) The ratio of the phenolic promoting solution to the control group (without the addition of the promoting solution) is the relative cell survival rate. Wherein, the cell line is selected as a glioma cell (gli〇ma C6, Food Industry Research Institute)' and the phosphate buffer solution is 气2g of potassium carbonate (KC1), 〇.24g of potassium dihydrogen phosphate ( Kh2P〇4), 8g of copper (NaCl) and 1.44g of disodium hydrogen phosphate (Na2HP〇4) are mixed and dissolved in 1L of deionized water (2d 0), and the pH is 7.2 to 7.4. PK10417 2007/8/20 200908954 Please refer to Fig. 2 again, which is the result of cell viability analysis of the carvacrol contained in the promoting liquid. From the results, it can be known that 'addition containing 〇~500 After the #M carvage stimulating solution, compared with the control group (c), 50% cell death was not achieved, so it was judged that carvacrol was not toxic to cells at 5 〇〇 #M or less. That is, the method of the present invention for promoting the production of an active oxide by a cell is not toxic to cells. Referring to Figures 3 and 4, the step of analyzing the active oxide production amount is: taking 1 ml of the cell liquid containing 5 x 105 cells (neurocolloma cells) per ml in I2well culture ιοί at 37 ° C And the cells were incubated in an incubator containing 5% carbon dioxide, followed by selective addition of lipopolysaccharide (LPS) at a concentration of 1 // g/ml, 400 ng/ml. 100 to 200 / / M carvacrol promoting solution, the lipopolysaccharide is a bacterial extract to simulate external microorganisms, which is used to increase the stimulation of the lipopolysaccharide, and then added after 30 minutes of action The concentration of 20 Μ dichlorodihydrofluorescein diacetate (DCFH-DA) (calbiochem) was selected for 30 minutes to obtain the dichlorofluorescein diacetate The active oxide produced by the cells binds and fluoresces, and then the RPMI culture solution is removed, washed once with the hydrochloride buffer, and then the phosphate buffer is removed, and then, 100//1 is added. Trypsin-EDTA (Gibco) to The captured cells from the dish, and then added to lml of the selected obstacle salt buffer broken cells, and finally a cell flow meter (fl〇w cytometry, Becton Dickinson; FACScan, PK10417 2007/8/20 -10 - 200908954
Calif〇raia)分析該細胞内自由基的產生量。 ，參照第3 _心其麵絲轉化物產生量分析 『九表現量相對細胞數之變倾。其中，第a組係為控 二’且’亦即該細胞液中並未添加脂多酿、τρΑ紐進液； :^之細胞液中係添加脂㈣及ΤΡΑ共同作用，並未添 口香/f酚；第c組之細胞液中僅添加含2⑻“Μ香芽酚之Calif〇raia) analyzes the amount of free radical production in the cell. , refer to the analysis of the amount of the third _ heart-shaped silk transformant. Among them, the first group is controlled by two 'and', that is, the cell fluid is not added with lipids, τρΑ New solution; : ^ cell liquid is added with lipid (four) and sputum, does not add a mouth /f phenol; only the 2 (8) "muxiang phenol" is added to the cell liquid of group c
促^液進行作用；帛d組之細胞液中係添加脂多聽、DA 膚香“之促進液共同仙，其巾，該脂多釀 ^ 為1/Zg/mi，該TPA之濃度係為40〇ng/ml，其他 ^ ]敘述於活性氧化物產生量分析之步驟中，於此不再 賢述。 )4再參照第3圖所示，由結果可得知，第&組與第。 、且、目車乂之下’可知於該細胞液中加人含2(Κ)"Μ香芽盼之 促進液料會觸發該細胞產生活性氧化物·，第&組與第b 、’且相車乂之下，可得知該細胞於脂多_及τρΑ存在之環境下 =生活性氧化物，⑼㈣b組與第cl組可發現，於脂 ^及TPA存在之環境下，加人含·" m香斧酴之促進 液明顯促進該細胞產生活性氧化物。駐所述，本發明之 =於該細胞未受外界微生物人侵時，將不會觸㈣細胞 產生活性氧化物’㈣免對該細胞造成傷害；反之，科 =受外界微生物人侵之環境下，力认含香_之促^ 將有效觸發該細胞產生較多量之活性氧化物 該外界微生物，進而提升該生物體之免疫力，藉！：： 明確實具有提升活性氧化物產生量及細胞免疫力之功效/ PK10417 2007/8/20 200908954 二芬如、第4圖所示，其係含不同香芽齡濃度之促進液 I X "^氧化物產生f分析之螢絲現量蝴細胞數之 义化圖。其中’第e組係為控制組’亦即該細胞液中並未 ，加月曰多_、TPA或促進液；第f組之細胞液中係添加脂 夕酿及TPA共同作用，並未添加該促進液；第g组之細胞 液中係添加脂多醣、TPA及含1〇〇// Μ之香序盼之促進液 共同作用；第h組之細胞液中係添加脂多酿、τρΑ及⑼ //M之香㈣共同作用；第ι組之細胞液中係、添加脂多酿 /PA及200//M之香芽龄共同作用，其中，該脂多聽之 濃度係為1 // g/ml ’該TPA之濃錢為彻_卜其他條 件則敘述於活性氧化物產生量分析之㈣巾，於此不再贅Promote the action of the liquid; in the cell liquid of the 帛d group, add the lipid to listen, the DA skin scent "the promoting liquid is common, and the towel, the fat is brewed ^ 1 / Zg / mi, the concentration of the TPA is 40〇ng/ml, other ^] is described in the step of analysis of the amount of active oxide production, which is not described here.) 4 Referring to Figure 3 again, the results can be known, the & group and And, under the eyelids, it is known that the addition of 2(Κ)" promoted liquids in the cell liquid triggers the cells to produce active oxides, the & groups and b Under the circumstance, it is known that the cell is in the environment of the presence of lipid _ and τρΑ = living oxide, (9) (4) b group and group cl can be found in the environment of lipid and TPA, plus The human-containing < m-astringent promoting solution significantly promotes the production of active oxides by the cells. In the present invention, the cells will not produce (IV) cells to produce active oxides when the cells are not invaded by external microorganisms. '(4) to avoid harm to the cell; on the contrary, the department = under the environment of external microbial invaders, the force of the fragrant _ will promote the cell production A large amount of active oxides of the external microorganisms, thereby enhancing the immunity of the organism, borrowed::: It has the effect of increasing the amount of active oxides produced and cellular immunity / PK10417 2007/8/20 200908954 As shown in Fig. 4, it is a map of the number of flutter cells in the IX "^ oxides containing different concentrations of germination aging. The 'e group is the control group' That is, the cell fluid is not added to the cellar _, TPA or promoting solution; the cell fluid of the f group is added with the lipid and TPA, and the promoting solution is not added; the cell liquid of the g group Adding lipopolysaccharide, TPA and a promoting solution containing 1〇〇// Μ 盼 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; In the cell liquid of the ι group, the addition of the fat poly/PA and the 200//M germination age, wherein the concentration of the lipid is 1 // g/ml 'The richness of the TPA is _ Other conditions are described in the analysis of the amount of active oxide production (four) towel, no longer 赘
述0 ^ 、曲請再參照第4圖所示，由結果可得知，隨著該香芽紛 濃度之增加’該活性氧化物之產生量亦有上升之趨勢。藉 此，可得知本發明之方法確實具有提升活性氧化物產生量及 細胞免疫力之功效。 …此外’ f更性肉芽腫病（—granul〇matous出獅&⑺〇 )係為-種遺傳性免疫缺陷疾病，主要是因為人體内白血球中 的吞嘆細胞有雜’細胞内之活性氧錄（例如··過氧化氯） 產生1不足，使得該吞噬細胞將微生物吞 :系频化步驟，而無法將病菌消滅’且容 =覆=: 感染，進而導致患者容易受到某些細菌與真菌的感染。另，肺 結核病係由結核桿菌入侵人體所狀，由於所施予之藥物 若無法持續且徹底的殺除結核桿菌，則肺結核病將不易痊 PK10417 2007/8/20 —12 200908954 爆。然而，本發明具有提升活性氧化物產生量及細皰免卢 之功效，因此，當該生物體患有慢性肉芽腫病或肺結核二曰士 ，本發明之方法可提升該生物體之細胞之活性氧化物產生旦才 進而提升細胞免疫力。亦即，本發明之促進細胞產生活性里草 化物之方法可應用於慢性肉芽腫病及肺結核病。 虱 如上所述，相較於習用殺菌劑僅適用於由密螺旋體屬 所造成之疾病，進而造成使用裕度低落之缺點；再者，該習用 殺菌劑僅係為一殺菌組成物，並未提供一促進細胞產生活性氧 化物之方法’使得該生物體内的活性氧化物產生量可能較為= 落。反觀，本發明係將一至少含有香序酚之促進液藉由一施 =方法施予一生物體之細胞，使該細胞於外界微生物入侵 時，可提升該細胞内活性氧化物之產生量。藉此，本發明 ϋ _實可提升使用裕度、活性氧化物產生量及細胞免疫力。 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脫離本發明之精神 和範圍之内，相對上述實施例進行各種更動與修改仍屬本 發明所保護之技術範疇，因此本發明之保護範圍當視後附 之申請專利範圍所界定者為準。 ΡΚ10417 2007/8/20 13 200908954 【圖式簡單說明】 第1圖··本發明之促進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 PK10417 2007/8/20 —14——Referring to Fig. 4, it can be seen from the results that as the concentration of the fragrant buds increases, the amount of active oxides also increases. From this, it is understood that the method of the present invention does have an effect of increasing the amount of active oxide produced and cellular immunity. ... In addition, 'f more granulomatosis (-granul〇matous lion & (7) 〇) is a hereditary immunodeficiency disease, mainly because the human cells in the white blood cells of the swallow cell have a 'intracellular reactive oxygen species Recording (for example, · chlorine peroxide) produces 1 deficiency, so that the phagocytic cells will swallow the microorganisms: the frequency of the step, and the disease can not be eliminated 'and the capacity = cover =: infection, which leads to patients susceptible to certain bacteria and fungi Infection. In addition, tuberculosis is caused by tubercle bacilli invading the human body. Tuberculosis will not be easy to kill if the drug is not sustained and completely eliminated. PK10417 2007/8/20 —12 200908954 Explosion. However, the present invention has an effect of increasing the amount of active oxide produced and the blistering effect. Therefore, when the organism has chronic granulomatosis or tuberculosis, the method of the present invention can increase the activity of the cells of the organism. Oxide production increases the cellular immunity. Namely, the method for promoting active cellulite production by the present invention can be applied to chronic granulomatous disease and tuberculosis.虱 As described above, compared with the conventional bactericide, it is only applicable to the disease caused by the genus Treponema, thereby causing the disadvantage of low use margin; further, the conventional bactericide is only a bactericidal composition, and is not provided. A method of promoting the production of active oxides by cells 'causes the amount of active oxide produced in the organism to be relatively low. In contrast, in the present invention, a promoting liquid containing at least a phenolic phenol is administered to a living body cell by a method, so that the cell can increase the amount of active oxide produced in the cell when it is invaded by an external microorganism. Thereby, the present invention can improve the use margin, the amount of active oxide generated, and the cellular immunity. While the invention has been described in connection with the preferred embodiments described above, it is not intended to limit the scope of the invention. The technical scope of the invention is protected, and therefore the scope of the invention is defined by the scope of the appended claims. ΡΚ10417 2007/8/20 13 200908954 [Simplified description of the drawings] Fig. 1 is a flow block diagram of a method for promoting the production of active oxides by cells of the present invention. Figure 2: Results of cell viability analysis of the present invention. Fig. 3 is a graph showing changes in the amount of fluorescence on the amount of active oxide produced by the present invention. Fig. 4 is a graph showing the change in the amount of fluorescent expression relative to the number of cells in the analysis of the amount of active oxide produced by the present invention under the promotion solution containing different concentrations of carvacrol. [Main component symbol description] S1 First step S2 Second step PK10417 2007/8/20 —14——