摘要
目的观察罗哌卡因对大鼠脂肪干细胞(ADSCs)增殖、凋亡及成骨分化的影响。方法分离并提取大鼠ADSCs,传代培养后,取对数生长期ADSCs分别加入含0 mg/mL、200 mg/mL、500 mg/mL罗哌卡因的培养基100µL,分别记为对照组、200 mg/mL组、500 mg/mL组。取各组细胞,继续培养24、48、72 h时,采用CCK-8实验观察细胞增殖能力。取各组细胞,继续培养24 h,使用流式细胞仪检测细胞凋亡情况。取各组细胞,加入成骨诱导培养基,观察各组细胞成骨分化能力,包括成骨潜能(以ALP活性表示)、矿化能力(以OD值表示)和成骨相关蛋白骨钙素(OCN)表达水平。结果与对照组相比,200 mg/mL组、500 mg/mL组细胞培养24、48、72 h时的增殖能力均显著下降,且500 mg/mL组低于200 mg/mL组。对照组、200 mg/mL组、500 mg/mL组细胞凋亡率分别为2.78%±0.23%、4.15%±0.62%、5.88%±0.78%,组间相比,P均<0.05。与对照组相比,200 mg/mL组、500 mg/mL组细胞ALP活性、矿化能力、OCN蛋白表达水平均显著下降,且500 mg/mL组低于200 mg/mL组。结论200 mg/mL、500 mg/mL的罗哌卡因均可抑制大鼠ADSCs增殖能力,降低其成骨能力,促进其凋亡,且500 mg/mL罗哌卡因的抑制效果高于200 mg/mL。
Objective To observe the effects of ropivacaine on proliferation,apoptosis and osteogenic differentiation of rat adipose-derived stem cells(ADSCs).Methods ADSCs were isolated and extracted from rats.After subculture,ADSCs in the logarithmic growth phase were added into medium containing 0,200 and 500 mg/mL ropivacaine of 100µL each,and were recorded as the control group,200 mg/mL group and 500 mg/mL group,respectively.Cells were taken from each group and cultured for 24,48 and 72 h.The proliferation ability of cells was observed by CCK-8 assay.Cells from each group were cultured for 24 h and apoptosis was detected by flow cytometry.The cells of each group were select-ed,osteogenic differentiation ability of cells in each group was observed by adding osteogenic induction medium,including osteogenic potential(indicated by ALP activity),mineralization ability(represented by OD value)and expression level of osteocalcin(OCN).Results Compared with the control group,the proliferation capacities of the 200 mg/mL group and 500 mg/mL group significantly decreased at 24,48 and 72 h,and that in the 500 mg/mL group was lower than that of the 200 mg/mL group.The apoptosis rates of the control group,200 mg/mL group and 500 mg/mL group were 2.78%±0.23%,4.15%±0.62%,and 5.88%±0.78%,respectively(all P<0.05).Compared with the control group,the ALP activity,mineralization ability and OCN protein expression levels in the 200 mg/mL group and 500 mg/mL group signifi-cantly decreased,and those were lower in the 500 mg/mL group than in the 200 mg/mL group.Conclusion Both 200 and 500 mg/mL ropivacaine can inhibit the proliferation of ADSCs,reduce their osteogenic ability and promote their apop-tosis,and the inhibitory effect of 500 mg/mL ropivacaine is higher than that of 200 mg/mL.
作者
王静
唐琳
宋双荣
苏小丽
任丽洁
王萍
WANG Jing;TANG Lin;SONG Shuangrong;SU Xiaoli;REN Lijie;WANG Ping(Department of Stomatology,Cangzhou Medical College,Cangzhou 061001,China)
出处
《山东医药》
CAS
2024年第9期52-55,共4页
Shandong Medical Journal
关键词
罗哌卡因
脂肪干细胞
细胞增殖
细胞成骨分化
细胞凋亡
ropivacaine
adipose-derived stem cells
cell proliferation
osteogenic differentiation
apoptosis
