注射用丹参多酚酸对周细胞增殖与迁移和凋亡的影响

Effect of Salvianolic acid for injection on the proliferation,migration and apoptosis of pericytes in vitro

目的探讨注射用丹参多酚酸对周细胞(C3H/10T1/2)增殖、迁移和凋亡的影响。方法C3H/10T1/2细胞采用氧糖剥夺/复氧(OGD/R)模型模拟缺血缺氧损伤。将细胞分为5组:正常组、模型组和低、中、高3个剂量的实验组(注射用丹参多酚酸的6.25,12.50,25.00μg·mL^-1),正常组将培养液置换为新鲜的高糖DMEM,6 h后敷箱培养18 h;模型组将培养液置换为无糖DMEM,6 h后换为高糖培养基敷箱培养18 h;实验组是在模型组的基础上6 h后置换为含药的高糖培养基培养箱孵育18 h。细胞计数法检测周细胞活力(OD值);细胞划痕实验检测周细胞迁移;蛋白质印迹法检测凋亡相关蛋白B淋巴细胞瘤-2(Bcl-2)、Bcl-2-相关X蛋白(Bax)的表达水平。结果正常组、模型组和低、中、高3个剂量的实验组的细胞活力分别是(100.00±8.18)%,(80.49±6.79)%,(98.36±9.65)%,(98.92±9.05)%和(103.10±8.73)%;这5组的细胞迁移率分别是(44.32±8.78)%,(26.96±2.78)%,(38.24±1.74)%,(41.13±1.88)%和(49.92±2.18)%;这5组的Bcl-蛋白表达水平分别是100%,(41.95±9.86)%,(109.73±9.00)%,(91.14±9.49)%和(82.14±10.36)%;这5组的Bax蛋白表达水平分别是100%,(131.61±4.60)%,(67.26±0.99)%,(85.17±14.35)%和(93.39±14.00)%。上述指标:模型组与正常组比较,或低、中、高3个剂量实验组与模型组比较,差异均有统计学意义(P<0.05,P<0.01)。结论注射用丹参多酚酸可以促进OGD/R损伤周细胞的细胞增殖及细胞迁移能力,抑制细胞凋亡。

Objective To investigate the effects of Salvianolic acid for injection on the proliferation,migration and apoptosis of pericytes(C3 H/10 T1/2).Methods Oxygen-glucose deprivation/reoxygenation(OGD/R)model was used to simulate ischemic and anoxic injury in cells.The cells were divided into 5 groups:normal group,model group and experimental-L,-M,-H groups(Salvianolic acid for injection:6.25,12.50,25.00μg·mL^-1).The normal group replaced the culture medium with fresh high Sugar DMEM,6 h later,cultured in the box for 18 h;the model group replaced the culture medium with sugar-free DMEM,after 6 h,changed to high-sugar medium,cultured in the box for 18 h;the experimental group was replaced with medicine-containing after 6 h on the basis of the model group incubate for 18 h in a high-sugar medium incubator.Cell counting kit-8 method was used to detect the proliferation level.The pericyte migration was detected by the cell scratch test.The expression levels of apoptosis-related proteins B cell lymphoma/lewkmia-2(Bcl-2)and Bcl-2-associated X protein(Bax)were detected by Western blot.Results The cell viability in normal group,model group,and low,medium,high dose experimental groups were(100.00±8.18)%,(80.49±6.79)%,(98.36±9.65)%,(98.92±9.05)%and(103.10±8.73)%,respectively;the migration rates in the 5 groups were(44.32±8.78)%,(26.96±2.78)%,(38.24±1.74)%,(41.13±1.88)%and(49.92±2.18)%,respectively;the expression level of Bcl-2 protein in the 5 groups were 100%,(41.95±9.86)%,(109.73±9.00)%,(91.14±9.49)%and(82.14±10.36)%;and the expression level of Bax protein in the 5 groups were 100%,(131.61±4.60)%,(67.26±0.99)%,(85.17±14.35)%and(93.39±14.00)%.Comparison between model group and normal group,or comparison between experimental-L,-M,-H groups and model group,the difference of the factors were significant(P<0.05,P<0.01).Conclusion Salvianolic acid for injection can promote the cell proliferation and cell migration ability of OGD/R injured pericytes and inhibit cell apoptosis.This study provides a basis for the clinical application of the protective effect of Salvianolic acid for injection on cerebral ischemia.