基于正交实验设计补阳还五汤水提液对高糖培养视网膜微血管内皮细胞活性影响  

Effect of Buyang Huanwu Decoction Water Extract on Activity of High Glucose CulturedRetinal Microvascular Endothelial Cells Based on Orthogonal Experimental Design

作　　者：曹俊昌 石颖 胡艳红[1] 胡俊[1] 陈胜[1] CAO Junchang;SHI Ying;HU Yanhong;HU Jun;CHEN Sheng(The Second Affiliated People's Hospital of Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350003,China)

机构地区：[1]福建中医药大学附属第二人民医院,福建福州350003

出　　处：《福建中医药》2023年第11期55-58,共4页Fujian Journal of Traditional Chinese Medicine

基　　金：福建省卫生健康科技计划项目医学创新课题(2020CXB034);福建省卫生健康科技计划项目青年科研课题(2022QNA080)。

摘　　要：目的基于正交实验筛选高糖损伤模型的最佳葡萄糖干预浓度、补阳还五汤水提液(BYHWT)保护高糖诱导视网膜微血管内皮细胞(HRCECs)损伤的最佳浓度及最佳作用时间。方法CCK8法检测4.5、5、10、15、20、25 mg/mL葡萄糖浓度和0、5、10、15.625、31.25、62.5、125、250 mg/mL BYHWT浓度对HRCECs细胞增殖的影响。采用L9(34)正交实验设计以葡萄糖浓度、BYHWT浓度及孵育时间为影响因素,以OD值为评价指标,优选出高糖损伤模型的最佳葡萄糖干预浓度、BYHWT防护高糖损伤HRCECs的最佳浓度及最佳作用时间。根据正交实验优选出的葡萄糖浓度和BYHWT浓度,将HRCECs细胞分为正常组、模型组(优选葡萄糖浓度)与干预组(优选葡萄糖浓度+优选BYHWT浓度),培养24、48、72 h,每日换液1次,CCK8法测定3组细胞不同时间的OD值。结果①与4.5 mg/mL组比较,25 mg/mL葡萄糖浓度干预HRCECs细胞24、48、72 h后OD值均明显降低(P<0.05),提示该浓度可引起细胞损伤。与0 mg/mL组比较,5 mg/mL BYHWT干预24 h,10、15.625 mg/mL BYHWT干预48、72 h后OD值均明显提高(P<0.05),提示该浓度可促进增殖。②筛选正交实验可知,影响细胞增殖活性的主次因素为孵育时间>葡萄糖浓度>BYHWT浓度,选择25 mg/mL葡萄糖浓度建立高糖损伤模型,选择5 mg/mL BYHWT浓度干预细胞,观察细胞增殖活性,筛选干预时间。③与正常组比较,模型组干预24、48、72 h后OD值明显降低(P<0.05);干预组干预24、48 h后OD值明显升高(P<0.05),干预72 h后明显降低(P<0.05)。与模型组比较,干预组干预24、48、72 h后OD值明显升高(P<0.05)。结论25 mg/mL葡萄糖培养的HRCECs细胞可以作为高糖损伤模型,5 mg/mL BYHWT干预24 h可以用于研究其对高糖诱导HRCECs损伤的保护作用。Objective:To select the optimal glucose intervention concentration for high glucose injury models based on orthogonal experiments,and to screen the optimal concentration and optimal incubation time of Buyang Huanwu decoction water extract(BYHWT)to protect high glucose induced damage to retinal microvascular endothelial cells(HRCECs).Methods:CCK8 was used to assess the effect of glucose concentrations(4.5,5,10,15,20,25 mg/mL)and BYHWT concentrations(0,5,10,15.625,31.25,62.5,125,250 mg/mL)on HRCECs proliferation.An L9(34)orthogonal experimental design was employed with glucose concentration,BYHWT concentration,and incubation time as influencing factors,and OD value as the evaluation index to select the optimal glucose intervention concentration,BYHWT concentration,and duration for preventing high glucose damage in HRCECs.HRCECs were divided into normal group,model group(optimal glucose concentration),and intervention group(optimal glucose concentration and BYHWT concentration).After 24,48,and 72 hours of cultivation,the CCK8 method was used to measure the OD value at different time points.Results:1)Compared with the 4.5 mg/mL group,the OD values of HRCECs significantly decreased after 24,48,and 72 hours of intervention with 25 mg/mL glucose concentration(P<0.05),indicating that this concentration can cause cell damage.Compared with the 0 mg/mL group,after 24 hours of intervention with 5 mg/mL BYHWT and 48 and 72 hours of intervention with 10 and 15.625 mg/mL BYHWT,the OD value significantly increased(P<0.05),indicating that this concentration can promote proliferation.2)According to orthogonal experimental design,the factors affecting cell proliferation activity were incubation time,glucose concentration,and BYHWT concentration.The 25 mg/mL glucose concentration was selected to establish the high glucose injury model,and the 5 mg/mL BYHWT concentration was chosen to intervene in cells to observe cell proliferation activity and screen intervention time.3)Compared with the normal group,the OD value of the model group

关 键 词：补阳还五汤 高糖 视网膜微血管内皮细胞 正交实验设计 

分 类 号：R285[医药卫生—中药学]