Han Siyin, Li Hongxuan, Luo Fei, Chen Xin, Cen Yanhui, Liu Peng, Chen Zhenxing, Lan Taijin, Lin Jiang
School of Basic Medicine, Guangxi University of Chinese Medicine, Nanning, China.
The First Clinical Medical College, Guangxi University of Chinese Medicine, Nanning, China.
Evid Based Complement Alternat Med. 2022 Jul 1;2022:1558288. doi: 10.1155/2022/1558288. eCollection 2022.
This study is an investigation into the inhibitory effect of seawater pearl hydrolysate (SPH) on the UVA-induced photoaging of human skin fibroblast (HSF) cells, and the mechanism thereof. HSF cells were cultured and irradiated with a UVA 0-50 J·cm dose gradient. The cell inhibition rate was detected using the CCK8 method, and the half-inhibitory dose was determined. Based on this, the dose of UVA irradiation for the follow-up experiment was selected to establish a photoaging model of the HSF cells. The cells were divided into a normal (N) group, UVA-irradiated (UVA) group, SPH low dose (SPHL) group, SPH medium dose (SPHM) group, and SPH high dose (SPHH) group. The photoaging model of HSF cells was established by UVA irradiation in the UVA, SPHL, SPHM, and SPHH groups; the SPHL, SPHM, and SPHH groups were treated with SPH at concentrations of 50, 100, and 200 mg·L, respectively, at the same time. After 24 and 48 h of culture, the reactive oxygen species (ROS) level of the HSF cells was detected by flow cytometry, and the required culture time of the HSF cells for the follow-up experiment was selected. The malondialdehyde and glutathione contents, as well as the activities of the superoxide dismutase, catalase, and glutathione peroxidase in the HSF cells, were detected by biochemical methods. The levels of expression of MMP-1 and collagen I protein in HSF cells were detected by the western blot test, the extent of aging of HSF cells was detected by -galactosidase staining, and the apoptosis level of HSF cells was detected by flow cytometry. The results show that SPH inhibits the UVA-induced photoaging of HSF cells in a dose-dependent manner within a certain concentration range, and the effect of a concentration of 200 mg·L was the most significant. The mechanism is related to improving the antioxidant activity of photoaging HSF cells to eliminate excessive ROS. It can inhibit apoptosis, reduce the protein expression of MMP-1, and effectively control the degradation of collagen I protein in photoaging HSF cells. Therefore, SPH offers potential for use in sunscreen cosmetics.
本研究旨在探讨海水珍珠水解物（SPH）对紫外线A（UVA）诱导的人皮肤成纤维细胞（HSF）光老化的抑制作用及其机制。培养HSF细胞，并用0 - 50 J·cm的UVA剂量梯度进行照射。采用CCK8法检测细胞抑制率，并确定半数抑制剂量。据此选择后续实验的UVA照射剂量，建立HSF细胞光老化模型。将细胞分为正常（N）组、UVA照射（UVA）组、SPH低剂量（SPHL）组、SPH中剂量（SPHM）组和SPH高剂量（SPHH）组。UVA、SPHL、SPHM和SPHH组通过UVA照射建立HSF细胞光老化模型；同时，SPHL、SPHM和SPHH组分别用浓度为50、100和200 mg·L的SPH进行处理。培养24和48小时后，通过流式细胞术检测HSF细胞的活性氧（ROS）水平，并选择HSF细胞后续实验所需的培养时间。采用生化方法检测HSF细胞中丙二醛和谷胱甘肽含量以及超氧化物歧化酶、过氧化氢酶和谷胱甘肽过氧化物酶的活性。通过蛋白质免疫印迹试验检测HSF细胞中基质金属蛋白酶-1（MMP-1）和I型胶原蛋白的表达水平，通过β-半乳糖苷酶染色检测HSF细胞的老化程度，通过流式细胞术检测HSF细胞的凋亡水平。结果表明，在一定浓度范围内，SPH以剂量依赖方式抑制UVA诱导的HSF细胞光老化，浓度为200 mg·L时效果最显著。其机制与提高光老化HSF细胞的抗氧化活性以消除过量ROS有关。它可以抑制细胞凋亡，降低MMP-1的蛋白表达，并有效控制光老化HSF细胞中I型胶原蛋白的降解。因此，SPH在防晒化妆品中具有应用潜力。
