摘要
目的:探讨当归红芪超滤物(UFE-AH)对X射线引起人脐静脉内皮细胞(HUVECs损伤的保护作用,阐明其可能的机制。方法:体外培养HUVECs,采用不同剂量(0、2、4、6、8和10 Gy) X射线辐射,筛选出最佳辐射剂量(6 Gy);用不同浓度(0、 50、100、200、400、600、800和1 000μg·L^(-1)) UFE-AH干预HUVECs,筛选出最佳促增殖浓度(100、200和400μg·L^(-1)^(-1))。实验分为空白组、模型组(6 Gy X射线)、低剂量UFE-AH组(6 Gy X射线+100μg·L^(-1)UFE-AH)、中剂量UFE-AH组(6 Gy X射线+200μg·L^(-1)UFE-AH)和高剂量UFE-AH组(6 Gy X射线+400μg·L^(-1)UFE-AH)。CCK-8法检测各组细胞存活率,流式细胞术检测各组细胞凋亡率,透射电子显微镜下观察各组细胞超微结构,Western blotting法检测各组细胞中磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B (Akt)、磷酸化Akt (p-Akt)、血管内皮生长因子(VEGF)、 B细胞淋巴瘤2 (Bcl-2)、Bcl-2相关X蛋白(Bax)和含半胱氨酸的天冬氨酸蛋白水解酶3 (caspase-3)蛋白表达水平。结结果果:CCK-8法检测,与0 Gy比较,当辐射剂量大于2 Gy时,辐射后48和72 h X射线对HUVECs的抑制率随辐射剂量增加而升高(P<0.05或P<0.01),6 Gy及6 Gy以上辐射剂量细胞抑制率升高更明显(P<0.01)。与0μg·L^(-1)UFE-AH比较,100、200、400和600μg·L^(-1)UFE-AH作用时细胞存活率升高(P<0.05或P<0.01),且100、200和400μg·L^(-1)UFE-AH作用24、48和72 h时细胞存活率明显升高(P<0.01)。与空白组比较,模型组细胞存活率明显降低(P<0.01);与模型组比较,中和高剂量UFE-AH组细胞存活率明显升高(P<0.01)。流式细胞术检测,与空白组比较,模型组细胞凋亡率明显升高(P<0.01);与模型组比较,低、中和高剂量UFE-AH组细胞凋亡率均明显降低(P<0.01)。透射电子显微镜观察,空白组细胞膜完整,胞质较均匀,线粒体呈卵圆形,未见明显肿胀,胞内可见少量溶酶体;与空白组比较,模型组细胞重度肿胀,细胞膜多处破损,胞质稀松且出现
Objective:To investigate the protective effect of ultra-filtration extract from Angelica Sinensis Radix and Hedysari Radix(UFE-AH)on the injury of human umbilical vein endothelial cells(HUVECs)induced by X-ray,and to clarify its possible mechanism. Methods:The HUVECs were cultured in vitro and irradiated with different doses(0,2,4,6,8 and 10 Gy)of X-ray;the optimal radiation dose(6 Gy)was selected. The HUVECs were intervened with different concentrations(0,50,100,200,400,600,800 and1 000 μg·L^(-1))of UFE-AH, and the optimal proliferation promoting concentrations(100,200 and400 μg·L^(-1))were selected. The experiment was divided into blank group,model group(6 Gy X-ray),low dose of UFE-AH group(6 Gy X-ray + 100 μg·L^(-1)UFE-AH),medium dose of UFE-AH group(6 Gy X-ray+200 μg·L^(-1)UFE-AH)and high dose of UFE-AH group(6 Gy X-ray + 400 μg·L^(-1)UFE-AH).The survival rates of cells in various groups were measured by CCK-8 assay,the apoptotic rates of cells in various groups were measured by flow cytometry,the ultrastructures were observed by transmission electron microscope,and the expression levels of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt),phosphorylated Akt(p-Akt),vascular endothelial growth factor(VEGF),B-cell lymphoma-2(Bcl-2),Bcl-2associated X protein(Bax)and cysteinyl aspartate specific proteinase-3(caspase-3)proteins in the cells in various groups were measured by Western blotting method. Results:The CCK-8 assay results showed that compared with 0 Gy X-ray,the inhibitory rates of HUVECs irradiated by X-ray were increased with the increase of radiation dose when the indiation dose was greater than 2 Gy at 48 and 72 h after radiation(P<0. 05 or P<0. 01),and the increasing of inhibition rates at 6 Gy X-ray and above were more significant(P<0. 01). Compared with 0 μg·L^(-1)UFE-AH,the survival rates of cells after treated with 100,200,400 and 600 μg·L^(-1)UFE-AH were increased(P<0. 05 or P<0. 01),especially treated with 100,200 and400 μg·L^(-1)UFE-AH for 24,48 and 72 h(P<0. 01). Compa
作者
武兵兵
张爱平
赵信科
李应东
刘凯
WU Bingbing;ZHANG Aiping;ZHAO Xinke;LI Yingdong;LIU Kai(Key Clinical Experimental Center of Integrated Traditional Chinese and Western Medicine,College of Integrated Chinese and Western Medicine,Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China;Department of Cardiovascular Medicine,Affiliated Hospital,Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2022年第5期1139-1147,共9页
Journal of Jilin University：Medicine Edition
基金
国家中医药管理局国家中医药循证能力建设项目(2019XZZX-XXG002)
甘肃省科技厅自然科学基金项目(21JR7RA562)
甘肃省中医药研究中心开放课题(ZYZX-2020-16)。
关键词
当归红芪超滤物
血管内皮细胞
细胞凋亡
X射线
辐射
放射治疗
Ultra-filtration extract from Angelica Sinensis Radix and Hedysari Radix
Vascular endothelial cells
Apoptosis
X-ray
Radiation
Radiation therapy
