The Protective Effects of Sodium Pentaborate Tetrahydrate Against UVB-induced Apoptosis in Human Keratinocytes

Ezgi Avşar Abdik
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Abstract

Ultraviolet radiation (UV) is an environmental carcinogen causing human skin cancer. Exposure of the skin to UV produces apoptotic keratinocytes called sunburn cells within the epidermis. Boron, an essential element for plants, has several biological properties, such as anti-cancer, anti-microbial, and anti-oxidant. In the present study, the possible protective effects of sodium pentaborate pentahydrate (SPT) against UVB-induced apoptosis in human keratinocyte cells, HaCaT, were investigated. They were treated with SPT at different concentrations (7.8-125 μg/mL) for 24h after UVB irradiation (20, 30 and 60mJ/cm2). Cell viability, annexin V assay, cell cycle analysis, and apoptosis-related gene levels were measured using RT-PCR. Treatment with SPT (15.6-31.25μg/mL) after 30 mJ/m2 UVB exposure significantly increased cell survival. Annexin V apoptosis analysis demonstrated a robust protective effect by treatment with SPT at concentrations of 15.6 and 31.25μg/mL after 30mJ/cm2 UVB irradiation. The cell cycle analysis revealed that UVB irradiation elevated the number of cells at the G0/G1 phase while SPT treatment after UVB irradiation increased the number of cells at G2/M phase, suggesting the changes were partially reversed. Furthermore, treatment with 15.6μg/mL SPT after 30 mJ/m2 UV irradiation blocked the activation of caspase 3, caspase 9, Bax, and p53. These results indicate that treatment with SPT exerts protective effects after UVB irradiation. Thus, treatment with SPT led to strong protection against UVB-induced apoptotic cell death in HaCaT cells.
四水五硼酸钠对uvb诱导的人角质形成细胞凋亡的保护作用
紫外线辐射是导致人类皮肤癌的环境致癌物。皮肤暴露在紫外线下会在表皮内产生凋亡的角质形成细胞,称为晒伤细胞。硼是植物必需元素,具有抗癌、抗微生物、抗氧化等多种生物学特性。在本研究中,研究了五水五硼酸钠(SPT)对uvb诱导的人角化细胞(HaCaT)凋亡的保护作用。在UVB(20、30和60mJ/cm2)照射后,分别给予不同浓度(7.8 ~ 125 μg/mL)的SPT处理24h。采用RT-PCR检测细胞活力、膜联蛋白V测定、细胞周期分析和凋亡相关基因水平。UVB照射30 mJ/m2后,SPT (15.6 ~ 31.25μg/mL)可显著提高细胞存活率。在30mJ/cm2的UVB照射下,15.6和31.25μg/mL浓度的SPT对膜联蛋白V的凋亡有显著的保护作用。细胞周期分析显示,UVB照射增加了G0/G1期细胞数量,而UVB照射后的SPT处理增加了G2/M期细胞数量,表明变化部分逆转。此外,在30 mJ/m2紫外线照射后,15.6μg/mL SPT可阻断caspase 3、caspase 9、Bax和p53的活化。这些结果表明,在UVB照射后,SPT治疗具有保护作用。因此,SPT治疗对uvb诱导的HaCaT细胞凋亡具有很强的保护作用。
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