葛根素对大鼠颈椎间盘纤维环细胞凋亡的影响  

Effects of puerarin on apoptosis of annulus fibrosis cells in rat cervical intervertebral disc

作　　者：何坚[1,2,3] 邱艺斌 罗庆禄[1,2,3] 

机构地区：[1]福建中医药大学康复医学院,福州350122 [2]福建省运动功能康复重点实验室,福州350003 [3]福建中医药大学附属康复医院,福州350003

出　　处：《中华中医药杂志》2015年第12期4509-4512,共4页China Journal of Traditional Chinese Medicine and Pharmacy

摘　　要：目的:观察不同浓度葛根素对白细胞介素-1β(IL-1β)诱导的大鼠颈椎间盘纤维环细胞凋亡率及凋亡因子B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)基因与胱天蛋白酶-3(Caspase-3)的影响。方法:对大鼠颈椎间盘纤维环细胞进行体外培养3代,鉴定成功后分为5组:对照组,模型组,葛根素低、中、高浓度组。除对照组外,其余4组均给予10ng/m L IL-1β诱导细胞凋亡,同时葛根素低、中、高浓度组分别予以125、250、500μmol/L的葛根素进行干预,24h后用流式细胞仪检测各组纤维环细胞凋亡率,Western blot检测凋亡调控基因Caspase-3和Bcl-2蛋白表达。结果:葛根素能抑制IL-1β诱导的椎间盘纤维环细胞凋亡。Caspase-3蛋白表达水平比较:与对照组比较,模型组显著升高(P<0.05);与模型组比较,3个给药组均显著降低(P<0.01,P<0.05),且随着葛根素浓度的提高表达减弱,葛根素高浓度组与葛根素低、中浓度组比较差异均有统计学意义(P<0.05)。3个给药组Bcl-2蛋白表达水平均显著高于对照组和模型组,差异具有统计学意义(P<0.01)。结论:一定浓度葛根素能够有效地抑制大鼠颈椎间盘纤维环细胞的凋亡,其途径可能是通过上调抗凋亡因子Bcl-2,下调促凋亡因子Caspase-3蛋白的表达。Objective： To explore the effects of different concentrations of puerarin on apoptosis rate, Bcl-2 gene and Caspase-3 gene of annulus fibrosis cells in rat cervical intervertebral disc induced by IL-1β. Methods： Isolated and cultured rat cervical intervertebral disc cells in vitro, stable three passaged, after identified cells were cultured successfully, then were randomly divided into control group, model group, low, middle and high content puerarin groups. All of the groups were intervened with 10ng/m L IL-1β except the control group in order to induce cervical intervertebral disc cells apoptosis. At the same time, low, middle and high content puerarin groups were intervened with 125, 250, 500μmol/L puerarin, respectively. After 24 h, l ow cytometry method was used to detect apoptosis of intervertebral disc cells, Western blot method was used to detected the protiens expression levels of Bcl-2 and Caspase-3 in different groups. Results： Puerarin could inhibit the apoptosis of intervertebral disc cells. The Caspase-3 expression in model group were singnificantly higher than control group（P〈0.05）, the Caspase-3 expression in all puerarin groups were singnificantly lower than model group（P〈0.01, P〈0.05）. With the concentrations puerarin increaseing, the Caspase-3 expression is decreasing, the caspase-3 expression in high content puerarin groups were singnificantly lowere than the low and middle content puerarin groups（P〈0.05）. The Bcl-2 expression in low, middle and high content puerarin groups were singnificantly higher than control group and model group（P〈0.01）. Conclusion： Puerarin could effectively inhibit apoptosis of annulus fibrosis cells in rat cervical intervertebral disc, which might be related to up-regulate anti-apoptotic factor Bcl-2 and downregulate the apoptotic factor Caspase-3.

关 键 词：葛根素 纤维环细胞 白细胞介素-1Β B淋巴细胞瘤-2基因 胱天蛋白酶-3 

分 类 号：R285[医药卫生—中药学]