康莱特对UVB照射后人皮肤成纤维细胞MMP1,MMP3的表达影响

发布时间：2018-03-04 09:00

本文选题：薏苡仁　切入点：基质金属蛋白酶　出处：《天津医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：本研究通过观察中药薏苡仁提取物康莱特(KLT)对中波紫外线(ultravioletB,UVB)照射的体外培养人皮肤成纤维细胞的基质金属蛋白酶1、3(Matrix Metallo Proteinases, MMP1、MMP3) mRNA及蛋白表达的影响,进一步探讨康莱特(KLT)是否抗光老化的药理作用。方法：采用酶消化法和组织块法培养人皮肤成纤维细胞；MTT法检测康莱特对人皮肽成纤维细胞增殖的影响；用不同辐照剂量UVB照射细胞后用实时荧光定量技术(RT-PCR)检测MMP1,MMP3mRNA表达,酶联免疫吸附试验(ELISA)法检测MMP1, MMP3蛋白水平；30mJ/cm2UVB照射细胞后分组分别加入1、2.5、5μL/mL康莱特基质及康莱特,用同上方法检测MMP1,MMP3mRNA及蛋白水平。结果：人皮肤成纤维细胞倍增时间较短,0-1d为潜伏期,1-3d为细胞增殖期,3-5d为对数生长期而5-7d为平台期。第6代之后细胞逐渐失去活力,生长缓慢,形态不规则。经MTT法检测加入不同浓度的康莱特细胞组结果显示与未加康莱特组比较,差异无统计学意义(p0.05)。30mJ/cm2UVB照射细胞组的MMP1, MMP3mRNA及蛋白表达水平与无UVB照射组的差异有统计学意义(p0.05)。30mJ/cm2UVB照射细胞组加入不同浓度基质及康莱特MMP1, MMP3mRNA及蛋白表达水平与对照组的差异有统计学意义(p0.05)。结论：人成纤维细胞可在较短时间内获得,第2代到第5代细胞增殖能力较强且生长稳定,宜用于实验。康莱特对人皮肤成纤维细胞的增殖无影响。康莱特可有效降低UVB照射后的人皮肤成纤维细胞MMP1, MMP3mRNA及蛋白的表达,且有剂量-效应关系。所以我们推测其有抗光老化的药理作用。
[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.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R285;R758.1

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