摘要
目的探讨益气化瘀方对H2O2诱导凋亡椎间盘纤维环细胞凋亡率的调控作用及作用机制。方法体外培养新西兰白兔纤维环细胞,RT-PCR技术检测纤维环细胞Ⅰ、Ⅱ型胶原及Aggrecan mRNA表达。在给予100μMH2O2诱导凋亡前1h、同时、延迟2h给予益气化瘀方药物血清,或中药干预同时给予PI3K/PKB信号通路阻断剂渥曼青霉素(Wortmannin),流式细胞仪检测不同时间点给药纤维环细胞凋亡率。结果与正常培养椎间盘纤维环细胞相比,100μM的H2O2诱导凋亡后,细胞凋亡率增高(P<0.01)。诱导凋亡前1h、同时、延迟2h给予益气化瘀方均可以降低凋亡率,诱导凋亡前或同时给予益气化瘀方,抗凋亡效果优于延迟给药;加入渥曼青霉素阻断PI3K/PKB信号通路后,益气化瘀方抗凋亡作用减弱,与同时加药组比较,差异有统计学意义(P<0.05)。结论益气化瘀方对H2O2诱导纤维化细胞凋亡有保护作用,其机理可能是通过激活PI3K信号转导通路,抑制纤维环细胞凋亡。
Objective To study the effect of "Yiqi Huayu Prescription" on apoptosis of annulus fibrosus cells induced by hydrogen peroxide(H2O2) and its mechanisms.Methods New Zealand white rabbit annulus fibrosus cells cultured in vitro were intervened by H2O2,with or without "Yiqi Huayu Prescription" at 1 hour before treatment,or the same time,or after 2 hours,furthermore,PI3K/PKB signaling pathway inhibitor wortmannin was added to "Yiqi Huayu Prescription" group.Cellular morphology was examined by light microscopy,ColleganⅠ,Ⅱand Aggrecan mRNA expressions were detected by RT-PCR technology;apoptotic changes were evaluated by flow cytometry.Results 100 μm H2O2 induced apoptotic changes in vitro.Treatment with "Yiqi Huayu Prescription" at different time all significantly reduced the rate of apoptosis,the effects of anti-appotosis of administration in advance and simultaneous administration was better than delayed administration.Furthermore,blocked PI3K/PKB signaling pathway,and the effect of "Yiqi Huayu Prescription" on anti-apoptosis decreased.Conclusion "Yiqi Huayu Prescription" protects annulus fibrosus cells from H2O2 induced apoptosis.The mechanism may activate the PI3K/PKB signaling pathway to promote cell survival annulus.
出处
《上海中医药杂志》
2010年第6期109-111,共3页
Shanghai Journal of Traditional Chinese Medicine
基金
国家杰出青年科学基金(30625043)
国家自然科学基金重大国际合作项目(30710103904)
国家自然科学基金青年基金(30801478
30701118
30600829)
上海市青年科技启明星计划(09QA1405600)
上海市优秀学科带头人计划(08XD1404000)
上海市教委创新项目(08YZ56)