补骨脂酚对光老化HSF细胞保护作用研究
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摘要
目的研究补骨脂酚对中波紫外线(UVB)照射诱导的光老化人皮肤成纤维细胞(HSF)的保护作用。方法体外培养HSF细胞,分为空白组(正常培养,不进行UVB辐射)、UVB模型组(UVB辐射)、雌二醇组及补骨脂酚低(1μmol/L)、中(10μmol/L)、高(100μmol/L)剂量组。采用MTT法检测细胞活力;采用Western blot法检测细胞内Ⅰ型胶原蛋白(COLⅠ)、基质金属蛋白酶-1(MMP-1)和基质金属蛋白酶-3(MMP-3)的表达量;采用逆转录PCR法(RT-PCR)检测细胞COLⅠ、MMP-1 mRNA的表达水平。结果确定光老化HSF细胞模型的最佳UVB照射剂量为25 mJ/cm2。与空白组比较,补骨脂酚低、中、高剂量组细胞活力及MMP-1、MMP-3、COLⅠ分泌差异无统计学意义(P> 0.05);UVB模型组的细胞活力、COLⅠ的表达水平显著下降(P <0.05),MMP-1和MMP-3的表达水平显著上升(P <0.05);MMP-1 mRNA表达水平升高、COLⅠmRNA表达水平下降(P <0.05)。与UVB模型组比较,补骨脂酚低、中、高剂量组细胞活力、COLⅠ的表达量显著上升(P <0.05),MMP-1和MMP-3的表达水平显著下降(P <0.05),MMP-1 mRNA表达下降、COLⅠmRNA表达上升(P <0.05)。结论补骨脂酚能够促进HSF细胞COLⅠ表达,抑制MMP-1与MMP-3的表达,防护UVB对HSF细胞的损伤,发挥抗皮肤光老化的作用。
Objective To study the protective effect of bakuchiol on photoaging human skin fibroblasts(HSF) induced by ultraviolet radiation B(UVB) irradiation. Methods HSF cells were cultured in vitro and divided into blank group(normal culture, no UVB radiation), UVB model group(UVB radiation), estradiol group and bakuchiol low(1 μmol/L),medium(10 μmol/L) and high dose(100 μmol/L) groups. Cell viability was detected by MTT assay. The protein expressions of type Ⅰ collagen(COL Ⅰ), matrix metalloproteinase-1(MMP-1) and matrix metalloproteinase-3(MMP-3)were detected by Western blot. The expression levels of COL Ⅰ and MMP-1 m RNA were detected by reverse transcription polymerase chain reaction(RT-PCR). Results The optimal UVB radiation dose for photoaging HSF cell model was 25 mJ/cm2. Compared with the blank group, there was no significant difference in cell viability and secretion of MMP-1, MMP-3 and COL Ⅰ between the low, medium and high dose groups of bakuchiol(P > 0.05); the cell viability and COL Ⅰ expression level in the UVB model group decreased significantly(P < 0.05), the protein expression levels of MMP-1 and MMP-3 increased significantly(P < 0.05); the m RNA expression levels of MMP-1 and COL Ⅰ decreased significantly(P < 0.05). Compared with the UVB model group, the cell viability and COL Ⅰ expression in the low, middle and high dose groups of bakuchiol increased significantly(P < 0.05), the protein expression levels of MMP-1 and MMP-3 decreased significantly(P <0.05), the mRNA expression levels of MMP-1 and COL Ⅰ increased significantly(P < 0.05). Conclusion Bakuchiol can promote the expression of COL Ⅰ in HSFs, inhibit the expression of MMP-1 and MMP-3,protect UVB from damage to HSFs and exert anti-photoaging effects on skin.
Objective To study the protective effect of bakuchiol on photoaging human skin fibroblasts(HSF) induced by ultraviolet radiation B(UVB) irradiation. Methods HSF cells were cultured in vitro and divided into blank group(normal culture, no UVB radiation), UVB model group(UVB radiation), estradiol group and bakuchiol low(1 μmol/L),medium(10 μmol/L) and high dose(100 μmol/L) groups. Cell viability was detected by MTT assay. The protein expressions of type Ⅰ collagen(COL Ⅰ), matrix metalloproteinase-1(MMP-1) and matrix metalloproteinase-3(MMP-3)were detected by Western blot. The expression levels of COL Ⅰ and MMP-1 m RNA were detected by reverse transcription polymerase chain reaction(RT-PCR). Results The optimal UVB radiation dose for photoaging HSF cell model was 25 mJ/cm2. Compared with the blank group, there was no significant difference in cell viability and secretion of MMP-1, MMP-3 and COL Ⅰ between the low, medium and high dose groups of bakuchiol(P > 0.05); the cell viability and COL Ⅰ expression level in the UVB model group decreased significantly(P < 0.05), the protein expression levels of MMP-1 and MMP-3 increased significantly(P < 0.05); the m RNA expression levels of MMP-1 and COL Ⅰ decreased significantly(P < 0.05). Compared with the UVB model group, the cell viability and COL Ⅰ expression in the low, middle and high dose groups of bakuchiol increased significantly(P < 0.05), the protein expression levels of MMP-1 and MMP-3 decreased significantly(P <0.05), the mRNA expression levels of MMP-1 and COL Ⅰ increased significantly(P < 0.05). Conclusion Bakuchiol can promote the expression of COL Ⅰ in HSFs, inhibit the expression of MMP-1 and MMP-3,protect UVB from damage to HSFs and exert anti-photoaging effects on skin.
引文
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[19]徐美娇.二苯乙烯苷对中波紫外线诱导人皮肤成纤维细胞光老化的保护作用研究[D].杭州:浙江中医药大学,2016.
[20]王继慧.绞股蓝总皂苷对人皮肤成纤维细胞光老化模型凋亡信号通路的影响[D].沈阳:辽宁中医药大学,2014.
[21]任捷.中草药对紫外线照射下真皮成纤维细胞保护作用的研究[D].上海:复旦大学,2010.
[2]杨锐,陈丽娟.中西医防治皮肤光老化新进展[J].中医药信息,2017,34(3):117-120.
[3]刘少英,孟祥璟,张祥奎,等.皮肤光老化机制及抗光老化药物[J].生理科学进展,2018,49(4):265-269.
[4]张昕,李承新.皮肤光老化治疗进展[J].中国激光医学杂志,2018,27(1):47-51.
[5]左俊,李勤,吴燕虹.雌激素调控皮肤衰老作用机制的研究进展[J].中国美容整形外科杂志,2013,24(2):122-124.
[6]张欢,李青峰.雌激素对皮肤衰老的调控[J].组织工程与重建外科,2009,5(1):58-60.
[7]雷秀东.雌激素抗皮肤老化的作用机制探讨[J].医学信息,2014(39):284-284.
[8]倪晨宁.皮肤光老化的中医药治疗研究进展[J].名医,2018(2):96,98.
[9] Antoniou C,Kosmadaki MG,Stratigos AJ,et al. Photoaging[J].Am J Clin Dermatol,2010,11(2):95-102.
[10] Zegarska B,Jó'z wicki W,Zegarski W,et al. Histopathological lesions in different types of skin aging[J]. Acta Dermatovenerol Alp Pannonica Adriat,2010,19(2):17-21.
[11] Chiu AE,Ab JLC,Ms DGK,et al. Double-Blinded,Placebo-Controlled Trial of Green Tea Extracts in the Clinical and Histologic Appearance of Photoaging Skin[J].Dermatol Surg,2005,31(Supplement s1):855-860.
[12] Jung HY,Shin JC,Park SM,et al. Pinus densiflora,extract protects human skin fibroblasts against UVB-induced photoaging by inhibiting the expression of MMPs and increasing typeⅠprocollagen expression[J]. Toxicol Rep,2014(1):658-666.
[13]刘俊岑,刘海洋,刘国良,等.异补骨脂素对紫外线诱导人真皮成纤维细胞损伤的保护作用及机制研究[J].国际中医中药杂志,2018,40(4):339-344.
[14]刘俊岑,董蕊,朴成玉,等.松脂醇二葡萄糖苷对UVB诱导HDF细胞损伤的保护作用及机制研究[J].中药材,2017,40(6):1410-1414.
[15]周勇,谢臻蔚,金杭美.雌激素在抗皮肤衰老中的生物学作用[J].中国美容医学,2017,26(1):27-30.
[16] Sjerobabskimasnec I,Situm M. Skin aging[J]. Acta Clinica Croatica,2010,49(4):515-518.
[17]魏毅君,张荣军,罗文颖,等.补骨脂酚通过抑制凋亡、氧化应激和炎症反应缓解小鼠脓毒症脑病[J].现代生物医学进展,2018,18(20):3840-3844.
[18] Xin Z,Wu X,Ji T,et al. Bakuchiol:A newly discovered warrior against organ damage[J]. Pharmacol Res,2019, 141.
[19]徐美娇.二苯乙烯苷对中波紫外线诱导人皮肤成纤维细胞光老化的保护作用研究[D].杭州:浙江中医药大学,2016.
[20]王继慧.绞股蓝总皂苷对人皮肤成纤维细胞光老化模型凋亡信号通路的影响[D].沈阳:辽宁中医药大学,2014.
[21]任捷.中草药对紫外线照射下真皮成纤维细胞保护作用的研究[D].上海:复旦大学,2010.