CN114796169A - 2-甲氧基萘在制备抗皮肤光损伤产品中的用途 - Google Patents
2-甲氧基萘在制备抗皮肤光损伤产品中的用途 Download PDFInfo
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Abstract
本发明属于生物医药技术领域,具体公开一种2‑甲氧基萘在制备抗皮肤光损伤产品中的用途。本发明实验结果证明,2‑甲氧基萘能够通过下调UVB诱导的氧化损伤细胞中MMP‑1蛋白的分泌水平、MMP‑1基因及MMP‑9基因的表达水平,并上调Ⅰ型胶原蛋白的分泌水平和COL1A1基因的表达水平来改善UVB诱导的细胞氧化损伤。
Description
技术领域
本发明属于生物医药技术领域,具体公开一种2-甲氧基萘在制备抗皮肤光损伤产品中的用途。
背景技术
皮肤是人体最大的器官,皮肤老化是身体老化的主要表现之一。紫外线导致的皮肤光老化是造成皮肤老化损伤最关键的因素。光老化皮肤表现为皮肤曝光部位粗糙、增厚、干燥,皮肤松弛、皱纹加深加粗,局部有过度的色素沉着或毛细血管扩张,甚至可能出现各种良性或恶性肿瘤(如日光角化病、鳞状细胞癌、恶性黑素瘤等)。
皮肤是人体的第一道防御屏障,覆盖全身。皮肤作为最大的器官,约占人体的16%。成人的皮肤面积约为1.2-2.0m2。皮肤与外界环境直接接触,具有感受外界刺激、调节体温、排泄代谢物、保护机体免受病原微生物的物理、机械、化学损伤和侵袭等功能。皮肤由表皮、真皮和皮下组织三部分组成。
紫外线是引起皮肤光老化的主要诱因。日光由53%的红外线、44%的可见光和3%的UV组成。UV是波长为100-400nm的电磁辐射。根据辐射波长的不同,UV可分为长波UV(UVA,波长为315-400nm)、中波UV(UVB,波长为280-315nm)和短波UV(UVC,波长为200-280nm)。适量的紫外线辐射可杀死微生物,调节神经、内分泌、消化、呼吸、免疫系统,促进维生素D的合成。但长期暴露于低剂量或瞬间暴露于高剂量紫外线辐射会对人体眼睛、皮肤、免疫系统等造成伤害。UVA和UVB是导致皮肤老化的主要光源。UVA导致细胞自由基生成和脂质过氧化的能力较强,可影响真皮组织中的胶原纤维和弹力纤维。由于UVA具有较强的穿透力,其影响可抵达真皮深层,虽对DNA损伤没有直接的影响,但可间接产生活性氧(reactive oxygenspecies,ROS),从而导致DNA氧化损伤。UVB主要引起皮肤表皮层和真皮浅层病变,UVB能被细胞中的蛋白质和DNA吸收,引起细胞损伤和变异。有研究表明,超过80%的面部皮肤老化是由紫外线照射造成的。皮肤光老化的宏观特征包括皱纹形成、质感粗糙、色素沉积、皮肤弹性丧失等;组织学和超微结构研究显示光老化皮肤表皮增生、胶原纤维受损紊乱、皮肤结缔组织中异常弹性物质大量积聚等。
2-甲氧基萘(2-Methoxynaphthalene),别名:2-萘甲醚、β-萘甲醚、甲基-β-萘基醚、橙花醚、乙位萘甲醚、甲基-2-萘基醚。2-甲氧基萘用于调配低档的有色皂、洗涤剂、日用品香精,适用香型为橙花、茉莉、百合。此外2-甲氧基萘还可用于低档的古龙和花露型香精中,可赋予较重的花香。但2-甲氧基萘在改善UVB诱导的细胞氧化损伤、抗皮肤光老化中的用途尚未见报道。
发明内容
鉴于以上技术问题,本发明提供以下技术方案:
本发明提供了2-甲氧基萘在制备抗皮肤光损伤产品中的用途。
优选地,所述2-甲氧基萘能够改善UVB诱导的细胞氧化损伤。
优选地,所述2-甲氧基萘能够下调MMP-1蛋白的分泌水平。
优选地,所述2-甲氧基萘能够上调Ⅰ型胶原蛋白的分泌水平。
优选地,所述2-甲氧基萘能够下调MMP-1基因及MMP-9基因的表达水平,并下调COL1A1基因的表达水平。
本发明还提供一种预防和/或治疗皮肤光损伤的药物组合物,其包括上述任一项所述的2-甲氧基萘或其盐,以及药学上可接受的辅料或载体。
本发明还提供一种改善皮肤光老化的保健食品组合物,其包括上述任一项所述的2-甲氧基萘或其盐,以及食品上可接受的辅料。
优选地,所述保健食品组合物能够改善皮肤皱纹、增强皮肤弹性、细化皮肤、减少色素沉积。
本发明还提供一种化妆品组合物,其包括上述任一项所述的2-甲氧基萘或其盐,以及至少一种化妆品上可接受的辅料或载体。
优选地,所述化妆品组合物能够改善紫外线照射引起的皮肤光老化和皮肤衰老。
对比现有技术,本发明的有益效果为:
本发明提供了2-甲氧基萘在制备抗皮肤光损伤产品中的用途。经细胞试验验证:2-甲氧基萘能够通过下调UVB诱导的氧化损伤细胞中MMP-1蛋白的分泌水平、MMP-1基因及MMP-9基因的表达水平,并上调Ⅰ型胶原蛋白的分泌水平和COL1A1基因的表达水平来改善UVB诱导的细胞氧化损伤,改善紫外线照射造成的皮肤皱纹形成、质感粗糙、色素沉积、皮肤弹性丧失等症状。
附图说明
图1是UVB辐照剂量对细胞存活率的影响;
图2是不同浓度的2-甲氧基萘对细胞存活率的影响;与0μM处理组相比较,**P<0.01;
图3是对UVB诱导的氧化损伤细胞中MMP-1蛋白分泌水平的影响;与UVB5组相比较,***P<0.001,****P<0.0001;
图4是对UVB诱导的氧化损伤细胞中PIP蛋白分泌水平的影响;与UVB5组相比较,**P<0.01,****P<0.0001;
图5是对UVB诱导的氧化损伤细胞中MMP-1基因表达水平的影响;与UVB5组相比较,*P<0.05;
图6是对UVB诱导的氧化损伤细胞中MMP-9基因表达水平的影响;与UVB5组相比较,**P<0.01;
图7是对UVB诱导的氧化损伤细胞中COL1A1基因表达水平的影响;与UVB5组相比较,*P<0.05,****P<0.0001。
具体实施方式
本发明结合实施例和相应附图做进一步阐释说明,以下实施例仅用于说明目的,不用于限制本发明范围。
本发明提供了2-甲氧基萘在制备抗皮肤光损伤产品中的用途。
上述2-甲氧基萘的结构式如式Ⅰ所示,分子式为C11H10O,相对分子质量158.2。
上述2-甲氧基萘的取得方法没有特别限制,可利用公知制法进行合成或从市场中购买得到。
上述2-甲氧基萘通过下调UVB诱导的氧化损伤细胞中MMP-1蛋白的分泌水平、MMP-1基因及MMP-9基因的表达水平,并上调Ⅰ型胶原蛋白的分泌水平和COL1A1基因的表达水平来改善UVB诱导的细胞氧化损伤。
下面结合具体实施例进行说明。
实施例1
诱导真皮成纤维细胞Hs 68光老化损伤
将Hs 68细胞接种在96孔板中,生长至密度50%时,给予0、3、5、7、10、20、30mJ/cm2的紫外辐照剂量,继续培养24小时后,用碧云天CCK8试剂盒,检测不同紫外辐照剂量下的细胞存活率。
实验结果(图1)显示,随着UVB辐照剂量增加,细胞的存活率逐渐减小。UVB辐照剂量以细胞存活率90%以上为宜,故选择UVB剂量为5mJ/cm2用于后续实验。
实施例2
2-甲氧基萘对Hs 68细胞的毒性
将Hs 68细胞接种在96孔板中,生长至密度50%时,给予0、12.5、25、50、100μM的2-甲氧基萘处理(以DMSO为溶剂),继续培养24小时后,用碧云天CCK8试剂盒,检测不同剂量下的细胞存活率。
根据图2结果,在实验浓度范围内2-甲氧基萘对Hs 68细胞均无毒性,所以选择实验范围内最大无毒性浓度100μM进行后续实验。
实施例3
2-甲氧基萘对MMP-1蛋白表达量的影响
将细胞接种在6孔板中,生长至密度50%时,分别给予UVB、UVB+浓度为100μM 2-甲氧基萘、UVB+0.1%DMSO处理,同时空白对照组细胞不做任何处理(标记为UVB0);各组继续培养24小时后,收集上清液和细胞。
MMP-1蛋白用elisa试剂盒检测,elisa检测原理:将MMP-1检测抗体通过物理吸附的方法固定于聚苯乙烯微孔板表面,加入待测样品,通过酶标物显色的深浅间接反映MMP-1含量。最后将MMP-1蛋白水平归一化至细胞总蛋白含量。
结果显示(图3),Hs 68细胞接受UVB辐照后,Hs 68细胞分泌的MMP-1蛋白水平极显著上升(****P<0.0001),而辐照后接受2-甲氧基萘干预可以极显著下调MMP-1蛋白的分泌水平(***P<0.001),溶剂DMSO对降低MMP-1蛋白分泌的效果无影响。
实施例4
2-甲氧基萘对Ⅰ型胶原蛋白分泌量的影响
将细胞接种在6孔板中,生长至密度50%时,分别给予UVB、UVB+浓度为100μM 2-甲氧基萘、UVB+0.1%DMSO处理,同时空白对照组细胞不做任何处理(标记为UVB0);各组继续培养24小时后,收集上清液和细胞。
Ⅰ型胶原蛋白是皮肤中最重要且含量最高的细胞外基质,皮肤中的Ⅰ型胶原蛋白由真皮成纤维细胞分泌。在细胞内合成的Ⅰ型前胶原分泌到细胞外,在肽链内切酶的作用下,其氨基端和羧基端附加的前肽被切断,形成原Ⅰ型胶原蛋白质,原胶原分子聚合成胶原纤维后构成细胞外基质。被切断的游离的前肽是可溶性的,可作为反映生物体内Ⅰ型胶原蛋白合成量的生物化学指标。可以利用I型前胶原C末端肽(简称PIP)特异性单克隆抗体对I型前胶原C末端肽进行定量。
结果显示(图4),Hs 68细胞接受UVB辐照后,Hs 68细胞分泌的PIP蛋白水平极显著下调(****P<0.0001),而辐照后接受2-甲氧基萘干预可以极显著升高PIP蛋白的分泌水平(**P<0.01),溶剂DMSO对升高PIP蛋白分泌的效果无影响。
实施例5
2-甲氧基萘对基质金属蛋白酶1基因(MMP-1)、基质金属蛋白酶9基因(MMP-9)及1型胶原α1链基因(COL1A1)相对表达量的影响
将Hs 68细胞接种在6孔板中,生长至密度50%时,分别给予UVB、UVB+浓度为100μM2-甲氧基萘处理、UVB+0.1%DMSO处理,同时空白对照组细胞不做任何处理(标记为UVB0);各组继续培养24小时后,收集细胞。
Real-time PCR分析:用Trizol试剂提取总RNA,按照逆转录试剂盒进行cDNA合成。实时荧光定量PCR实验使用iQ SYBR green supermix(Bio-Rad).。PCR扩增后以人β-actin或GAPDH为内参计算目的基因MMP-1、MMP-3及MMP-9的相对表达量(CFX ConnectTMReal-Time PCR Detection System,Bio-Rad,Hecules,CA,USA)。
图5显示,Hs 68细胞接受UVB辐照后,胞内MMP-1基因表达水平显著上调(*P<0.05),而辐照后接受2-甲氧基萘干预,MMP-1基因表达水平显著下降(*P<0.05),溶剂DMSO对降低MMP-1基因的表达水平的效果无影响。
图6显示,Hs 68细胞接受UVB辐照后,胞内MMP-9基因表达水平极显著上调(**P<0.01),而辐照后接受2-甲氧基萘干预,MMP-9基因表达水平有降低,但未达到统计学差异,溶剂DMSO对降低MMP-9基因的表达水平的效果无影响。
图7显示,Hs 68细胞接受UVB辐照后,胞内COL1A1基因表达水平显著下降(****P<0.0001),而辐照后接受2-甲氧基萘干预,COL1A1基因表达水平上升(*P<0.05),溶剂DMSO对提高COLIA1基因的表达水平的效果无影响。
本发明并不限于上述实施方式,在不背离本发明的实质内容的情况下,本领域技术人员可以想到的任何变形、改进、替换均落入本发明的范围。
Claims (10)
1.2-甲氧基萘在制备抗皮肤光损伤产品中的用途。
2.根据权利要求1所述的用途,其特征在于,所述2-甲氧基萘能够改善UVB诱导的细胞氧化损伤。
3.根据权利要求2所述的用途,其特征在于,所述2-甲氧基萘能够下调MMP-1蛋白的分泌水平。
4.根据权利要求2所述的用途,其特征在于,所述2-甲氧基萘能够上调Ⅰ型胶原蛋白的分泌水平。
5.根据权利要求2所述的用途,其特征在于,所述2-甲氧基萘能够下调MMP-1基因及MMP-9基因的表达水平,并上调COL1A1基因的表达水平。
6.一种预防和/或治疗皮肤光损伤的药物组合物,其特征在于,其包括权利要求1-5任一项所述的2-甲氧基萘或其盐,以及药学上可接受的辅料或载体。
7.一种改善皮肤光老化的保健食品组合物,其特征在于,其包括权利要求1-5任一项所述的2-甲氧基萘或其盐,以及食品上可接受的辅料。
8.根据权利要求7所述的保健食品组合物,其特征在于,所述保健食品组合物能够改善皮肤皱纹、增强皮肤弹性、细化皮肤、减少色素沉积。
9.一种化妆品组合物,其特征在于,其包括权利要求1-5任一项所述的2-甲氧基萘或其盐,以及至少一种化妆品上可接受的辅料或载体。
10.根据权利要求9所述的化妆品组合物,其特征在于,所述化妆品组合物能够改善紫外线照射引起的皮肤光老化和皮肤衰老。
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