本文選題：歐前胡素 + 異歐前胡素　； 參考：《瀘州醫(yī)學(xué)院》2013年碩士論文

【摘要】：目的：通過體外建立模擬正常生理狀態(tài)的人表皮黑素細(xì)胞培養(yǎng)體系，觀察白芷的兩種主要單體歐前胡素、異歐前胡素和白芍的主要單體芍藥苷對黑素細(xì)胞生物活性及遷移的影響，從而探討這三種中藥單體對黑素細(xì)胞的作用機(jī)制，為色素性疾病的治療提供新的思路。方法：1、體外培養(yǎng)模擬皮膚正常生理狀態(tài)的人表皮黑素細(xì)胞；2、檢測不同濃度的三種中藥單體（歐前胡素、異歐前胡素、芍藥苷）對黑素細(xì)胞增殖活性的影響（MTT法）；3、檢測不同濃度的三種中藥單體（歐前胡素、異歐前胡素、芍藥苷）對黑素細(xì)胞酪氨酸酶活性的影響（L-DOPA法）；4、檢測不同濃度的三種中藥單體（歐前胡素、異歐前胡素、芍藥苷）對黑素細(xì)胞黑素合成的影響（NaOH法）；5、采用Transwell小室檢測三種中藥單體（歐前胡素、異歐前胡素、芍藥苷）對黑素細(xì)胞遷移的影響。以上全部數(shù)據(jù)采用SPSS統(tǒng)計(jì)軟件17.0版進(jìn)行分析。結(jié)果：1、歐前胡素組在濃度為20μg/mL和50μg/mL時；異歐前胡素組在濃度為5μg/mL、10μg/mL、20μg/mL時；芍藥苷組在濃度為5μg/mL、10μg/mL、20μg/mL、50μg/mL時均顯示出顯著的促進(jìn)黑素細(xì)胞增殖的作用。各組中藥單體在100μg/mL時并未對黑素細(xì)胞增殖產(chǎn)生明顯的抑制作用，細(xì)胞仍顯示較好的生長。各組中藥單體對黑素細(xì)胞增殖的促進(jìn)作用均是低濃度時呈濃度依賴性增強(qiáng)，而較高濃度時反而作用力度下降，由實(shí)驗(yàn)結(jié)果可見，10μg/mL為芍藥苷和補(bǔ)骨脂素的最佳作用濃度；20μg/mL為歐前胡素和異歐前胡素的最佳作用濃度。其中5μg/mL、10μg/mL、20μg/mL濃度時的異歐前胡素組促細(xì)胞增殖能力強(qiáng)于同濃度的補(bǔ)骨脂素組。2、歐前胡素組在濃度為20μg/mL、50μg/mL時；異歐前胡素組在濃度為10μg/mL、20μg/mL時；芍藥苷組在10μg/mL、20μg/mL、50μg/mL濃度時；均顯示出上調(diào)黑素細(xì)胞酪氨酸酶活性的作用，并且其作用強(qiáng)度與同濃度的補(bǔ)骨脂素組相似。各組中藥單體對調(diào)節(jié)黑素細(xì)胞酪氨酸酶活性的作用也呈現(xiàn)出低濃度時劑量依賴性上調(diào)，而高濃度時上調(diào)能力逐漸下降。10μg/mL為異歐前胡素和芍藥苷的最佳作用濃度；20μg/mL為歐前胡素和補(bǔ)骨脂素的最佳作用濃度。3、歐前胡素組在濃度為2.5μg/mL、5μg/mL、10μg/mL、20μg/mL、50μg/mL、100μg/mL時均未顯示出對黑素合成的促進(jìn)作用；異歐前胡素組僅在20μg/mL時顯著促進(jìn)黑素合成；芍藥苷組在濃度為20μg/mL、50μg/mL時顯著的促進(jìn)黑素合成。異歐前胡素組、芍藥苷組、補(bǔ)骨脂素組促進(jìn)黑素合成的作用均是低濃度時劑量依賴性上調(diào)，高濃度時下降。20μg/mL為異歐前胡素和補(bǔ)骨脂素的最佳作用濃度；50μg/mL為芍藥苷的最佳作用濃度。4、10μg/mL、20μg/mL、50μg/mL的歐前胡素組均無明顯促進(jìn)黑素細(xì)胞遷移的作用；異歐前胡素組在這三個濃度時促黑素細(xì)胞遷移的作用呈現(xiàn)濃度依賴性加強(qiáng)，并于20μg/mL和50μg/mL時顯示出極顯著的促進(jìn)黑素細(xì)胞遷移的作用，與空白對照組比較在統(tǒng)計(jì)學(xué)上具有顯著性差異（P0.01）；芍藥苷組在20μg/mL時極顯著的促進(jìn)黑素細(xì)胞遷移，與空白對照組比較在統(tǒng)計(jì)學(xué)上具有顯著性差異（P0.01）；補(bǔ)骨脂素組（陽性對照組）在10μg/mL、20μg/mL、50μg/mL三個濃度均顯著的促進(jìn)黑素細(xì)胞遷移，其中20μg/mL時其作用強(qiáng)度最高；50μg/mL濃度組的異歐前胡素和補(bǔ)骨脂素均極顯著的促進(jìn)黑素細(xì)胞遷移，與空白對照組比較在統(tǒng)計(jì)學(xué)上具有顯著性差異（P0.01），并且此時兩者作用強(qiáng)度相當(dāng)。結(jié)論：1、歐前胡素、異歐前胡素、芍藥苷均能促進(jìn)黑素細(xì)胞的增殖，且在較高濃度（100μg/mL）時對細(xì)胞無明顯毒性作用，證實(shí)了這三種中藥單體的安全性。并且異歐前胡素促黑素細(xì)胞增殖活性能力強(qiáng)于補(bǔ)骨脂素。2、歐前胡素、異歐前胡素、芍藥苷均能上調(diào)黑素細(xì)胞酪氨酸酶的活性，，且作用強(qiáng)度與補(bǔ)骨脂素相似。3、異歐前胡素和芍藥苷均能促進(jìn)黑素細(xì)胞的黑素合成，而歐前胡素未檢測出促進(jìn)黑素合成的作用。4、20μg/mL、50μg/mL組的異歐前胡素和20μg/mL組的芍藥苷均能顯著地促進(jìn)黑素細(xì)胞遷移，而10μg/mL、20μg/mL、50μg/mL濃度組的歐前胡素均無明顯促進(jìn)黑素細(xì)胞遷移的作用，由此推測異歐前胡素單體是白芷中促進(jìn)黑素細(xì)胞遷移的主要活性成分，芍藥苷單體是白芍中促進(jìn)黑素細(xì)胞遷移的主要活性成分。50μg/mL組的異歐前胡素和補(bǔ)骨脂素促黑素細(xì)胞遷移的能力相當(dāng)。
[Abstract]:Objective: To observe the effects of two main monomeric monosonoids, isomer and Paeoniae paeoniflorin on the bioactivity and migration of melanocytes by establishing a human epidermal melanocyte culture system in vitro to simulate normal physiological state, and to explore the mechanism of action of these three kinds of monomers on melanocytes. The treatment of vegetative diseases provides new ideas. Methods: 1, human epidermal melanocytes are cultured in vitro to simulate the normal physiological state of the skin. 2, the effects of three kinds of traditional Chinese medicine monomers on the proliferation of melanocytes (MTT method); 3, three different concentrations of traditional Chinese medicine (erhu) The effect of peptide, ISO, paeoniflorin on the activity of tyrosinase in melanocytes (L-DOPA); 4, the effects of three kinds of Chinese herbs on melanin synthesis of melanocytes (NaOH method) were detected by different concentrations of Chinese herbs (eomerhu, ISO, and paeoniflorin); 5, using the Transwell chamber to detect three kinds of Chinese medicine monomers (eomerhu, isooin, peony, Paeonia lactiflora The effect of glucoside on melanocyte migration. All the above data were analyzed by the 17 edition of SPSS software. Results: 1, when the concentration was 20 mu g/mL and 50 mu g/mL, the concentration of ISO euclixin group was 5 u g/mL, 10 mu g/mL, 20 mu g/mL, and the concentration of paeoniflorin in the concentration was 5 mu g /mL, 10 u g/mL, 20 u g/mL, 50 g/mL. The proliferation of melanocytes was promoted. The cell proliferation of melanocytes was not inhibited significantly at 100 g/mL. The cells still showed good growth. The experimental results showed that 10 mu g/mL was the best concentration of paeoniflorin and psoralen; 20 g/mL was the best concentration of eomorsin and isooctin. The proliferation ability of ISO euclixin group at 5 mu g/mL, 10 mu g/mL and 20 mu g/mL was stronger than that of the same concentration of.2 in the same concentration of bonalin group, and the concentration of oealin group was 20 mu g/mL, 50 mu. At the time of g/mL, when the concentration was 10 mu g/mL and 20 u g/mL, the group of paeoniflorin showed the role of up regulation of tyrosinase activity in melanocytes at 10 u g/mL, 20 mu g/mL and 50 micron g/mL, and its action intensity was similar to that of the same concentration of psoralen group. At low concentration, the dose dependence was also up, while the up-regulation ability of the high concentration decreased gradually.10 mu g/mL as the best concentration of isoeuin and paeoniflorin; 20 micron g/mL was the best concentration of.3, and the oealin group was 2.5 u g/mL, 5 u g/mL, 10 mu g/mL, 20 mu g/mL, 50 u g/mL, 100 micron g/mL. The promoting effect of melanin synthesis was not shown; the group of isooctin was significantly promoted by melanin synthesis at 20 g/mL; paeoniflorin group promoted melanin synthesis at a concentration of 20 mu and 50 mu g/mL. The effect of the group of isoeuin, paeoniflorin group, and psoralen group on melanin synthesis was up to a dose dependent up and high concentration in low concentration. The decrease of.20 mu g/mL was the best concentration of isoeuin and psoralen, the best concentration of 50 mu g/mL was.4,10 mu g/mL, 20 g/mL, and 50 uh g/mL had no significant effect on the migration of melanocytes; the effect of the migration of melanocytes at these three concentrations showed a concentration dependent manner. The effect on melanocyte migration was significantly increased at 20 g/mL and 50 u g/mL, which was significantly different from that in the blank control group (P0.01); the group of paeoniflorin significantly promoted the migration of melanocytes at 20 u g/mL, and had significant differences compared with the blank control group (P0.01). Psoralen group (positive control group) significantly promoted melanocyte migration at 10 g/mL, 20 g/mL and 50 micron g/mL, of which 20 mu g/mL had the highest strength, and the 50 mu g/mL concentration group of isooctin and psoralen all significantly promoted melanocyte migration, compared with the blank control group, the difference was statistically significant difference. The difference (P0.01), and at the same time, has the same action intensity. Conclusion: 1, eulein, isoeulin and paeoniflorin all can promote the proliferation of melanocytes and have no obvious toxic effects on the cells at a high concentration of 100 mu, which confirms the safety of the three monomers of the Chinese medicine. Psoralen.2, ocuarinin, isooctin and paeoniflorin can all up regulate the activity of melanocyte tyrosinase, and the action intensity is similar to.3, isooctin and paeoniflorin all can promote melanin synthesis in melanocytes, but the effect of eucartin on melanin synthesis is not detected in.4,20 mu g/mL, and the 50 mu g/mL group of isooeus Paeoniflorin in group 20 and 20 mu could significantly promote melanin cell migration, while 10 mu g/mL, 20 mu g/mL, and 50 mu g/mL concentration group did not significantly promote melanocyte migration. Thus, it is suggested that ISO oculin monomer is the main active component in the promotion of melanocyte migration in Angelica dahurica, and paeoniflorin monomer is the promoting melanin in Paeonia lactiflora The main active components of cell migration were isoisoaltein and Psoralen in.50 g/mL group.

【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R758.4;R392.12

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