柴达木黑果枸杞花青素减轻中波紫外线辐射后人皮肤成纤维细胞氧化及炎性损伤的研究  

Study on alleviating the oxidative and inflammatory damage of HSFs cells from UVB radiation induced by the anthocyanins in Lycium ruthenicum from Chaidamu

作　　者：李喃欣 燕华玲[1] 余亚利 王丽雯[1] 张钦宁 万光梅 张美红 哈筱梅[1] LI Nanxin;YAN Hualing;YU Yali;WANG Liwen;ZHANG Qinning;WAN Guangmei;ZHANG Meihong;HA Xiaomei(Department of Dermatology,Qinghai University Affiliated Hospital,Xining 810000,China;Shijingshan Teaching Hospital,Capital Medical University,Beijing 100043,China)

机构地区：[1]青海大学附属医院皮肤科,青海西宁810000 [2]首都医科大学石景山教学医院,北京100043

出　　处：《西部医学》2023年第4期501-505,共5页Medical Journal of West China

基　　金：青海省科技厅项目(2019-ZJ-7058)。

摘　　要：目的 探讨柴达木黑果枸杞花青素减轻中波紫外线(UVB)辐射后人皮肤成纤维细胞(HSFs)氧化及炎性损伤。方法 将体外培养的HSFs细胞采用随机数字表法分为空白对照组(不进行任何处理)、UVB辐射组(30 mJ/cm~2照射30 min)、UVB+花青素低剂量组(0.1 mg/mL+30 mJ/cm~2照射30 min)、UVB+花青素中剂量组(0.5 mg/mL+30 mJ/cm~2照射30 min)、UVB+花青素高剂量组(1 mg/mL+30 mJ/cm~2照射30 min)。柴达木黑果枸杞花青素于UVB照射前24 h加入细胞培养基中,24 h后收集细胞。采用倒置显微镜观察细胞形态;台盼蓝染色观察细胞死亡情况;MTT法测细胞增殖活力;酶标法检测细胞超氧化物歧化酶(SOD)活力、过氧化氢酶(CAT)活力、谷胱甘肽(GSH-PX)含量、丙二醛(MDA)的含量。采用酶联免疫吸附实验(ELISA)法检测并比较白介素1(IL-1)、白介素6(IL-6)表达水平。结果 倒置显微镜下各组细胞形态:花青素中、高剂量组细胞形态接近于空白对照组,花青素低剂量组细胞形态接近于UVB副射组。台盼蓝染色显示,与空白对照组比较,UVB辐射组HSFs细胞蓝染数量增加,并出现漂浮细胞碎片;与UVB辐射组比较,UVB+低、中、高剂量柴达木黑果枸杞花青素组HSFs细胞蓝染数量下降,漂浮的细胞碎片减少。与空白对照组比较,UVB辐射组MDA、IL-1、IL-6表达水平明显增高(P<0.05);细胞增殖率、SOD活性、CAT活力、GSH-P含量下降,差异具有统计学意义(P<0.05)。与UVB辐射组比较,UVB+低、中、高剂量柴达木黑果枸杞花青素组细胞上清液中IL-1、IL-6、MDA表达水平明显降低(P<0.05);细胞增殖率、SOD活性、CAT活力、GSH-P含量均升高,差异具有统计学意义(P<0.05)。结论 柴达木黑果枸杞花青素通过提高抗氧化应激能力、减轻细胞的炎性损伤来改变UVB辐射后HSFs细胞的形态学变化,从而减轻紫外线辐射后引起的皮肤损伤。Objective To investigate the effect of anthocyanins in Lycium ruthenicum from Chaidamu on the antioxidative and inflammatory injury of human skin fibroblasts(HSFs cells)after UVB radiation.Methods HSFs cells cultured in vitro were randomly divided into 5 groups by random number table method:blank control group(no treatment),UVB irradiation group(30mJ/cm 2 irradiation for 30 minutes),UVB+low-dose anthocyanin group(0.1 mg/mL+30 mJ/cm 2 irradiation for 30 minutes),UVB+medium-dose anthocyanin group(0.5 mg/mL+30 mJ/cm 2 irradiation for 30 minutes),UVB+high-dose anthocyanin group(1 mg/mL+30 mJ/cm 2 irradiation for 30 minutes).24 hours before UVB radiation,the anthocyanins in Lycium ruthenicum from Chaidamu were added into the culture medium,and cells were collected after 20 hours radiation.The morphology of cells was observed by inverted microscope,cell death was observed by Trypan Blue.Cell proliferation activity was detected by MTT.SOD activity,CAT activity,GSH-Px content and MDA content was tested by enzyme linked immunosorbent assay.The expression levels of interleukin-1(IL-1),interleukin-6(IL-6)were detected and compared by enzyme-linked immunosorbent assay(ELISA).Results Trypan Blue showed that the number of blue staining of HSFs cells increased and floating cell fragments appeared in UVB radiation group compared with blank control group.Compared with UVB radiation group,the number of blue staining of HSFs cells and floating cell fragments decreased in UVB+low,medium and high dose anthocyanin group.The expression levels of MDA,IL-1 and IL-6 in UVB radiation group were significantly higher than those in blank control group(P<0.05).The cell proliferation rate,SOD activity,CAT activity,GSH-P content decreased,and the differences were statistically significant(P<0.05).Compared with UVB radiation group,the expression levels of IL-1,IL-6 and MDA in cell supernatant decreased significantly(P<0.05).The cell proliferation rate,SOD activity,CAT activity,GSH-P content increased,and the difference was statistically significa

关 键 词：柴达木黑果枸杞花青素 人皮肤成纤维细胞 中波紫外线辐射 氧化损伤 白介素1 白介素6 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学]