發(fā)布時(shí)間：2018-03-19 09:23

  本文選題：中藥　切入點(diǎn)：中藥色素　出處：《湖南中醫(yī)藥大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：1.研究背景與意義色素異常性皮膚病可分為色素增加性皮膚病(如黃褐斑)和色素減退性皮膚病(如白癜風(fēng))兩大類,其發(fā)病率呈不斷上升趨勢(shì)。由于色素異常性皮膚病主要影響人的外觀,因此給患者造成較大的社會(huì)壓力,亦明顯降低患者的生活質(zhì)量。色素異常性疾病的具體發(fā)生原因尚未明確,但與多種因素相關(guān),如遺傳、免疫、紫外線照射和內(nèi)分泌失調(diào)等。西醫(yī)藥治療色素異常性皮膚病藥物種類較多,費(fèi)用高且療程長(zhǎng),給患者帶來(lái)較大的精神壓力和經(jīng)濟(jì)負(fù)擔(dān)。中醫(yī)藥在治療色素異常性皮膚病的過(guò)程中積累了豐富的經(jīng)驗(yàn),歐陽(yáng)恒教授創(chuàng)立“以色治色”療法,并研發(fā)創(chuàng)制紫銅消白方(紫銅、丹參、紫草、紅花等多味紫紅色藥材)治療白癜風(fēng)獲得了較好的臨床療效,并榮獲1994年度國(guó)家中醫(yī)藥管理局科技進(jìn)步獎(jiǎng)。有些學(xué)者考慮中藥本身的顏色與色素異常性皮膚病的治療存在一定相關(guān)性,認(rèn)為深色類中藥對(duì)治療白癜風(fēng)等色素障礙性皮膚病有良好療效,淺色類中藥對(duì)治療黃褐斑等色素增加性皮膚病療效頗佳。楊柳教授[6]把這種表明藥材顏色與疾病療效相關(guān)的現(xiàn)象稱之為“中藥色象”。那么,單純的中藥的色素成分與色素異常性皮膚病之間是否存在直接相關(guān)性,目前尚未十分明確,相關(guān)研究報(bào)道亦偏少。因此,中藥色素成分與色素異常性皮膚病之間的相關(guān)性是一個(gè)值得研究的問題。2.研究目的:2.1通過(guò)實(shí)驗(yàn)研究證明中藥色素成分與人體皮膚色素代謝之間的相關(guān)性,直接證明中醫(yī)“以色治色”理論的正確性,并提供有力的理論依據(jù)和實(shí)驗(yàn)依據(jù)。2.2為中醫(yī)藥治療疾病提供新的治療方向和研究領(lǐng)域,促進(jìn)新藥研發(fā)。2.3發(fā)掘出天然、綠色、無(wú)毒害的色素,拓展中藥色素的應(yīng)用范圍,產(chǎn)生理想的社會(huì)效益和經(jīng)濟(jì)效應(yīng)。3.研究方法3.1查閱關(guān)于治療白癜風(fēng)、黃褐斑等色素異常性皮膚病的中醫(yī)藥研究類文獻(xiàn),篩選出常用于治療色素異常性皮膚病的中藥材,根據(jù)中藥外觀的顏色將中藥分為深色組(黑色、褐色、紫色、紅色)和淺色組(白色、黃色)。在深色類中藥組和淺色類中藥組的每種色系中分別選取2種目前成分研究較為明確的中藥色素進(jìn)行實(shí)驗(yàn)研究。3.2以中藥色素干預(yù)細(xì)胞,采用現(xiàn)代研究技術(shù)較為成熟的MTT法檢測(cè)中藥色素對(duì)細(xì)胞增殖的影響,多巴氧化法檢測(cè)中藥色素對(duì)酪氨酸酶活性的影響和NaOH裂解法測(cè)定其對(duì)黑素含量生成的影響。3.3統(tǒng)計(jì)學(xué)處理:運(yùn)用配伍組設(shè)計(jì)方法,色素為處理因素,濃度為配伍因素。實(shí)驗(yàn)結(jié)果中計(jì)量資料以“均數(shù)標(biāo)±準(zhǔn)差“(x±s)表示,采用SPSS19.0軟件包進(jìn)行統(tǒng)計(jì)分析。進(jìn)行單應(yīng)變量方差分析的最小顯著差異法(LSD)檢驗(yàn)。4.研究結(jié)果4.1梔子黃色素作用于小鼠B16黑素瘤細(xì)胞的安全濃度為4mg/ml。4.2梔子黃色素對(duì)細(xì)胞增殖的影響實(shí)驗(yàn)結(jié)果12h:與陰性對(duì)照組相比,梔子黃色素在安全濃度范圍內(nèi)(0.25mg/ml~4mg/ml)對(duì)細(xì)胞增殖無(wú)明顯抑制作用;24h:梔子黃色素溶液濃度在0.25mg/ml~1mg/ml范圍內(nèi)對(duì)細(xì)胞增殖無(wú)明顯抑制作用;濃度在2mg/ml~4mg/ml范圍內(nèi)對(duì)細(xì)胞增殖有較明顯的抑制作用,其抑制作用隨梔子黃色素溶液濃度的增大而增加;48h:梔子黃色素溶液濃度為0.25mg/ml時(shí)對(duì)細(xì)胞增殖無(wú)明顯作用;濃度在0.5mg/ml~4mg/ml范圍內(nèi)對(duì)細(xì)胞增殖有明顯抑制作用,其抑制作用隨梔子黃色素溶液濃度的增大而增加。4.3梔子黃色素對(duì)小鼠B16黑素瘤細(xì)胞酪氨酸酶活性影響梔子黃色素溶液在濃度為0.25mg/ml時(shí)對(duì)酪氨酸酶活性無(wú)明顯作用,在0.5mg/ml~4mg/ml濃度范圍內(nèi)時(shí)對(duì)酪氨酸酶活性有明顯的抑制作用。4.4梔子黃色素對(duì)小鼠B16黑素瘤細(xì)胞黑素生成影響梔子黃色素溶液在濃度為0.25mg/ml時(shí),對(duì)黑素生成無(wú)明顯抑制作用。梔子黃色素溶液在濃度0.5mg/ml~4mg/ml范圍內(nèi)時(shí),對(duì)黑素生成具有明顯抑制作用。4.5梔子黃色素對(duì)細(xì)胞外酪氨酸酶活性影響梔子黃色素溶液各濃度組OD值與陰性對(duì)照組OD值相比,其差異有明顯統(tǒng)計(jì)學(xué)意義(P0.01)表明梔子黃色素對(duì)蘑菇酪氨酸酶的活性有顯著的抑制作用,且隨著梔子黃色素濃度的增加,酪氨酸酶活性抑制率增大,測(cè)定的IC50分別為50mg/ml,100mg/ml。結(jié)論1.細(xì)胞實(shí)驗(yàn)1.1梔子黃色素作用于小鼠B16黑素瘤細(xì)胞的安全濃度為4mg/ml。1.2梔子黃色素在12h時(shí)間點(diǎn)對(duì)細(xì)胞增殖無(wú)明顯抑制作用,在24h、48h時(shí)間點(diǎn),能夠抑制小鼠B16黑素瘤細(xì)胞的增殖,其抑制作用呈濃度依賴性,即隨梔子黃色素溶液濃度的增大其抑制作用越明顯。1.3梔子黃色素能夠抑制小鼠B16黑素瘤細(xì)胞酪氨酸酶活性,其抑制作用呈濃度依賴性,即隨梔子黃色素溶液濃度的增大其抑制作用越明顯。1.4梔子黃色素能夠抑制黑素生成,其抑制作用呈濃度依賴性,即隨梔子黃色素溶液濃度的增大其抑制作用越明顯。2.細(xì)胞外實(shí)驗(yàn)梔子黃色素對(duì)蘑菇酪氨酸酶的活性有顯著的抑制作用,且隨著梔子黃色素濃度的增加,酪氨酸酶活性抑制率增大,測(cè)定的IC50分別為50mg/ml,100mg/ml。
[Abstract]:1. research background and significance of abnormal pigment skin disease can be divided into hyperpigmentation disease (such as chloasma) and hypopigmented skin (vitiligo) two categories, the incidence rate showed a rising trend. Due to abnormal pigmentation skin disease mainly affects the appearance of people, thus causing great social pressure to patients were also significantly lower the quality of life of patients. The specific causes of pigment abnormalities is not yet clear, but with many factors, such as genetic, immune, ultraviolet radiation and endocrine disorders. Western medicine treatment of pigmented skin disease abnormal many drugs, high cost and long course of treatment, great mental pressure and the economic burden for patients. Traditional Chinese medicine has accumulated rich experience in the treatment of abnormal pigmentation of skin diseases, Professor Ouyang Heng founded "by the color for color therapy, and created research white copper, copper consumption (Salvia miltiorrhiza Shikonin, safflower, multi flavor purple herbs) in the treatment of vitiligo has better clinical efficacy, and won the 1994 annual National Administration of traditional Chinese medicine science and technology progress award. Some scholars consider the treatment of traditional Chinese medicine itself abnormal color and pigment skin diseases there is a certain correlation, that the dark kind of traditional Chinese medicine has a good effect on the treatment of vitiligo. Pigmented skin disorder. The light of Chinese traditional medicine on the treatment of chloasma and hyperpigmentation disease has good curative effect. It shows that the medicine Professor Yang Liu [6] color associated with disease phenomenon known as the "Chinese medicine color like. So, whether there is a direct correlation between pigment pigment composition and abnormal skin disease pure Chinese medicine at present, is not clear, relevant research reports are few. Therefore, the correlation between the traditional Chinese medicine pigment pigment composition and abnormal skin disease is a problem worthy of study.2 Objective: 2.1. Through the experimental study proved the correlation between traditional Chinese medicine and pigment composition of human skin pigment metabolism, direct proof of correctness of traditional Chinese medicine to cure color color "theory, and provides a strong theoretical basis and experimental basis for.2.2 provides a new treatment direction and research in the field of Chinese medicine for the treatment of diseases, promote new drug research and development.2.3 discover the natural, green, non-toxic pigment, expand the scope of application of traditional Chinese medicine pigment, social benefit and economic effect of the.3. generation of ideal 3.1 consulting on the treatment of vitiligo, traditional Chinese medicine research literature of pigment chloasma and other abnormal skin disease, were often used in the treatment of pigmented skin disease abnormal herbs according to the traditional Chinese medicine, Chinese medicine will be divided into the appearance of the color of dark (black, brown, purple group, red light) and group (white, yellow). In the dark class Chinese medicine group and Chinese medicine group each kind of light color In traditional Chinese medicine were selected 2 kinds of components of pigment is clear for experimental study of.3.2 with traditional Chinese medicine intervention by pigment cells, modern research technology influencing pigment MTT assay on cell proliferation of mature Chinese medicine, traditional Chinese medicine pigment DOPA oxidation detection effect on tyrosinase activity and cleavage of NaOH method for the determination of its impact on the formation of melanin content.3.3 statistics processing: using the design method of compatibility group, pigment treatment, concentration of compatibility factors. The experimental results of the measurement data to mean + standard deviation standard "(x + s), analyzed by SPSS19.0 software. The single variable ANOVA least significant difference method (LSD) the results of.4. test in the safe concentration of B16 murine melanoma cells 4.1 Gardenia Yellow Gardenia Yellow effect of 4mg/ml.4.2 on cell proliferation effect of experimental results of 12h: and negative control Compared the gardenia yellow pigment in the safe concentration (0.25mg/ml~4mg/ml) had no significant inhibitory effect on the proliferation of 24h: cells; gardenia yellow pigment concentration in the range of 0.25mg/ml~1mg/ml on cell proliferation is not inhibited; the concentration in the range of 2mg/ml~4mg/ml on cell proliferation was restrained significantly, with the inhibition of gardenia yellow pigment solution the concentration increases with the increase of the concentration of 48h: solution; gardenia yellow is no obvious effect on the proliferation of 0.25mg/ml cells; the concentration has obvious inhibitory effects on cell proliferation in the range of 0.5mg/ml~4mg/ml, the inhibitory effect of gardenia yellow pigment with the concentration increasing and the increase of.4.3 of gardenia yellow on mice B16 melanoma cells tyrosinase activity effect of gardenia yellow pigment solution at the concentration of 0.25mg/ml had no obvious effect on tyrosinase activity, 0.5mg/ml~4mg/ml concentration in the range of tyrosine The enzyme activity significantly inhibited.4.4 of gardenia yellow on melanogenesis in B16 melanoma cells of gardenia yellow pigment solution at the concentration of 0.25mg/ml, generation of black pigment is not inhibited. Gardenia yellow pigment solution concentration in the range of 0.5mg/ml~4mg/ml, the melanogenic has obvious inhibitory effect on.4.5 of gardenia yellow the extracellular activity of tyrosinase effect of Gardenia yellow pigment solution with different concentration group was compared with the negative control group OD value, there was significant difference (P0.01) showed that the inhibition activity of gardenia yellow on mushroom tyrosinase is significant, and with the increasing concentration of gardenia yellow, tyrosinase activity inhibition rate increase, determination of IC50 respectively. For 50mg/ml, the safe concentration in B16 murine melanoma cells 100mg/ml. cells in experiment 1.1 conclusion 1. Gardenia Yellow Gardenia Yellow role for 4mg/ml.1.2 in 12h Time no obvious inhibition on cell proliferation in 24h, 48h time point, can inhibit the proliferation of B16 murine melanoma cells, the inhibitory effect was concentration dependent, with the increase of solution concentration of gardenia yellow pigment in the more obvious inhibition of gardenia yellow.1.3 can inhibit B16 melanoma cells tyrosinase activity in mice and its inhibition in a concentration dependent manner, with the gardenia yellow pigment concentration increased more obvious inhibition of gardenia yellow.1.4 can inhibit melanin production, its inhibitory effect in a concentration dependent manner, the inhibition increased significantly inhibited the more obvious.2. cell experiment of gardenia yellow on mushroom tyrosinase activity with gardenia yellow in the solution concentration, and with the increasing concentration of gardenia yellow, tyrosinase activity inhibition rate increase, determination of IC50 were 50mg/ml, 100mg/ml.

【學(xué)位授予單位】：湖南中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R285.5

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