本文選題：薏苡仁　切入點：基質(zhì)金屬蛋白酶　出處：《天津醫(yī)科大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：本研究通過觀察中藥薏苡仁提取物康萊特(KLT)對中波紫外線(ultravioletB,UVB)照射的體外培養(yǎng)人皮膚成纖維細(xì)胞的基質(zhì)金屬蛋白酶1、3(Matrix Metallo Proteinases, MMP1、MMP3) mRNA及蛋白表達(dá)的影響,進(jìn)一步探討康萊特(KLT)是否抗光老化的藥理作用。 方法：采用酶消化法和組織塊法培養(yǎng)人皮膚成纖維細(xì)胞；MTT法檢測康萊特對人皮肽成纖維細(xì)胞增殖的影響；用不同輻照劑量UVB照射細(xì)胞后用實時熒光定量技術(shù)(RT-PCR)檢測MMP1,MMP3mRNA表達(dá),酶聯(lián)免疫吸附試驗(ELISA)法檢測MMP1, MMP3蛋白水平；30mJ/cm2UVB照射細(xì)胞后分組分別加入1、2.5、5μL/mL康萊特基質(zhì)及康萊特,用同上方法檢測MMP1,MMP3mRNA及蛋白水平。 結(jié)果：人皮膚成纖維細(xì)胞倍增時間較短,0-1d為潛伏期,1-3d為細(xì)胞增殖期,3-5d為對數(shù)生長期而5-7d為平臺期。第6代之后細(xì)胞逐漸失去活力,生長緩慢,形態(tài)不規(guī)則。經(jīng)MTT法檢測加入不同濃度的康萊特細(xì)胞組結(jié)果顯示與未加康萊特組比較,差異無統(tǒng)計學(xué)意義(p0.05)。30mJ/cm2UVB照射細(xì)胞組的MMP1, MMP3mRNA及蛋白表達(dá)水平與無UVB照射組的差異有統(tǒng)計學(xué)意義(p0.05)。30mJ/cm2UVB照射細(xì)胞組加入不同濃度基質(zhì)及康萊特MMP1, MMP3mRNA及蛋白表達(dá)水平與對照組的差異有統(tǒng)計學(xué)意義(p0.05)。 結(jié)論：人成纖維細(xì)胞可在較短時間內(nèi)獲得,第2代到第5代細(xì)胞增殖能力較強且生長穩(wěn)定,宜用于實驗�？等R特對人皮膚成纖維細(xì)胞的增殖無影響。康萊特可有效降低UVB照射后的人皮膚成纖維細(xì)胞MMP1, MMP3mRNA及蛋白的表達(dá),且有劑量-效應(yīng)關(guān)系。所以我們推測其有抗光老化的藥理作用。
[Abstract]:Objective: to observe the effects of Coix seed extract (KLTT) on the expression of matrix metalloproteinase (MMP1) Metallo protein 3 (MMP1MMP3) mRNA and protein in cultured human skin fibroblasts irradiated with UVB. To further investigate the pharmacological effect of Kanglaite KLTs on photoaging. Methods: the effect of Kanglaite on the proliferation of human dermal fibroblasts was detected by enzyme digestion and tissue mass method, and the expression of MMP1 and MMP3 mRNA was detected by real-time fluorescence quantitative technique (RT-PCR) after irradiation with different doses of UVB. The protein level of MMP1 / MMP3 was detected by Elisa (Elisa). The cells were irradiated with 30 mJ / cm ~ (-2) UVB. The cells were treated with 12. 5 渭 L / mL Conlet matrix and Kanglaite respectively. The mRNA and protein levels of MMP1 / MMP3 were detected by the same method. Results: the multiplication time of human skin fibroblasts was shorter than 0-1d, the incubation period was 1-3d, the proliferative phase was 3-5d and the logarithmic growth stage was 5-7d. After the sixth passage, the cells gradually lost their activity and grew slowly. The morphology was irregular. The results of different concentrations of Kanglaite cell group were compared with those without Kanglaite group by MTT assay. There was no significant difference in the expression of MMP1, MMP3mRNA and protein between the UVB group and the control group. There was a significant difference in the expression of MMP1, MMP3mRNA and protein between the UVB group and the control group. There was a significant difference in the expression of MMP1, MMP3mRNA and protein between the UVB group and the control group. There was a significant difference in the expression of MMP1, MMP3mRNA and protein between the UVB group and the control group. The difference was statistically significant (P 0.05). Conclusion: human fibroblasts can be obtained in a short time. Kanglaite has no effect on the proliferation of human skin fibroblasts. Kanglaite can effectively reduce the expression of MMP1, MMP3mRNA and protein in human skin fibroblasts irradiated with UVB. And there is a dose-effect relationship, so we speculate that it has anti-photoaging pharmacological effect.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R285;R758.1

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