神经肽P物质对放射损伤皮肤成纤维细胞周期及凋亡的影响

摘要/Abstract

摘要： 为探讨神经肽P物质(substance P,SP)对放射损伤皮肤成纤维细胞凋亡及周期的影响,将人皮肤成纤维细胞(human foreskin fibroblasts,HFF-1)分为0 Gy组(对照组)和12 Gy组、18 Gy组、12 Gy+SP组、18 Gy+SP组4个实验组。实验组经12 Gy、18 Gy的电子束照射,其中12 Gy+SP组、18 Gy+SP组于照射前1 h给予10^-7 mol/L SP干预。照射后48 h,流式细胞术检测细胞周期及细胞凋亡率;实时荧光定量PCR检测Bcl-2、Bax表达情况。细胞周期检测显示,与0 Gy组相比,12 Gy和18 Gy组细胞G₂期百分比显著增加;18 Gy+SP组与18 Gy组相比,细胞G₂期百分比显著降低。细胞凋亡检测显示,与0 Gy组相比,12 Gy和18 Gy组细胞凋亡率显著升高;SP干预组(12 Gy+SP组、18 Gy+SP组)较照射组(12 Gy组、18 Gy组)的细胞凋亡率显著降低。实时荧光定量PCR对Bcl-2和Bax表达检测显示,与0 Gy组相比,12 Gy和18 Gy照射组细胞Bax基因表达显著升高,Bcl-2基因表达显著降低;SP干预组(12 Gy+SP组、18 Gy+SP组)较照射组(12 Gy组、18 Gy组),Bcl-2基因表达显著升高,Bax基因表达差异不显著。上述结果揭示,电子束照射可诱发人皮肤成纤维细胞的凋亡及细胞G₂期阻滞;SP可抑制放射损伤人皮肤成纤维细胞的凋亡及G₂期阻滞。

关键词: 神经肽P物质, 人皮肤成纤维细胞, 放射损伤, 细胞凋亡, 细胞周期

Abstract: To explore the effects of substance P on cell cycle and apoptosis of irradiated skin fibroblasts, Human Foreskin Fibroblasts (HFF-1) in logarithmic growth phase were divided into five experimental groups: 0 Gy group, 12 Gy group, 18 Gy group, 12 Gy+SP group and 18 Gy+SP group. The experimental groups were irradiated with electron beam at doses of 12 Gy and 18 Gy, while the 12 Gy+SP group and 18 Gy+SP group were treated with 10^-7 mol/L SP one hour before irradiation. After 48 hours of irradiation, cell cycle and apoptosis were detected by flow cytometry. The expression of apoptosis-related genes Bcl-2 and Bax in cells of each experimental group was quantitatively detected by real-time quantitative PCR. Cell cycle detection showed that the percentage of cells in G₂ phase in 12 Gy and 18 Gy experimental groups was significantly higher than that in 0 Gy group. And, compared with the 18 Gy experimental group, the percentage of cells in G₂ phase in the 18 Gy+SP experimental group was significantly lower than that in the 18 Gy group. Apoptosis detection showed that compared with 0 Gy experimental group, the apoptosis rate of 12 Gy and 18 Gy irradiation group increased significantly, while the apoptosis rate of substance P intervention group (12 Gy+SP, 18 Gy+SP) was significantly lower than that of radiation group (12 Gy, 18 Gy). Real-time fluorescence quantitative PCR detection of Bcl-2 and Bax expression showed that compared with 0 Gy experimental group, Bax gene expression was significantly increased and Bcl-2 gene expression was significantly decreased in 12 Gy and 18 Gy irradiated groups. And, the expression of Bcl-2 gene in substance P intervention group (12 Gy+SP and 18 Gy+SP) was significantly higher than that in radiation group (12 Gy and 18 Gy), but there was no significant difference in Bax gene expression. The above results suggest that electron irradiation can induce apoptosis and cell G₂ phase arrest of human skin fibroblasts. Substance P can inhibit the apoptosis and G₂ phase arrest of human skin fibroblasts damaged by radiation.

Key words: substance P, human skin fibroblasts, radiation injury, cell apoptosis, cell cycle

中图分类号:

刘晓明, 党旭红, 张睿凤, 李晓臻, 原雅艺, 刘红艳, 柴栋良, 任越, 张忠新, 左雅慧. 神经肽P物质对放射损伤皮肤成纤维细胞周期及凋亡的影响[J]. 辐射防护, 2023, 43(1): 83-89.

LIU Xiaoming, DANG Xuhong, ZHANG Ruifeng, LI Xiaozhen, YUAN Yayi, LIU Hongyan, CHAI Dongliang, REN Yue, ZHANG Zhongxin, ZUO Yahui. Effects of substance P on cell cycle and apoptosis of irradiated skin fibroblasts[J]. RADIATION PROTECTION, 2023, 43(1): 83-89.

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