健脾补土法组方含药血清对体外嗅鞘细胞低氧/复氧损伤模型P65、P50及磷酸化IκBα表达的影响

Effect of Spleen-Strengthening Therapy Contained Serum on P65, P50 and P-IκBα Expression in Olfactory Ensheathing Cells with Hypoxia / Reoxygenation Injury Models

目的探讨健脾补土法组方含药血清对体外嗅鞘细胞低氧/复氧损伤模型核因子KB(nuclear factor-kappa B,NFKB)的亚单位P65、P50及NF-KB抑制蛋白(P-IκBα)表达的影响。方法将原代培养的嗅鞘细胞分为5组:正常血清对照组、正常血清+低氧组、低氧+依达拉奉含药血清组、低氧+健脾补土法组方含药血清组、正常血清+低氧+NF-KB抑制剂/抗氧化剂(PDTC,50μmol/L)组,采用微需氧袋低氧/复氧的方法诱导嗅鞘细胞形成脑缺血再灌注损伤样模型,用Western blot法检测嗅鞘细胞胞浆P65、P50、P-IκBα和胞核P65、P50的含量。结果与正常血清对照组比较,正常血清+低氧组胞浆和胞核P65、P50,胞浆P-IκBα表达均显著升高,差异有统计学意义(P<0.01);与正常血清+低氧组比较,低氧+依达拉奉组、低氧+健脾补土组、正常血清+低氧+PDTC组、低氧+健脾补土+PDTC组胞浆和胞核P65、P50,胞浆P-IκBα表达均降低,差异有统计学意义(P<0.05)。结论健脾补土法组方能通过抑制嗅鞘细胞缺氧/复氧损伤后P65、P50、P-IκBα的过度表达,对NF-κB通路的活化具有抑制作用。

Objective To investigate the effect of spleen-strengthening therapy contained serum on subgene P65, P50 of nudear factor-kappa B （NT-KB） and P-IκBα expression in olfactory ensheathing cells with hypoxia/reoxygenation injury. Methods Primary cultured olfactory ensheathing cells were randomly divided into five groups： normal serum control group, normal serum＋hypoxia group, hypoxia＋edaravone contained serum group, hypoxia＋spleen-strengthening therapy contained serum group, normal serum＋hypoxia＋NF-KB inhibitor/antioxidant （PDTC 50 μmol/L） group. The cerebral ischemia-reperfusion injury model of olfactory ensheathing cells were induced by hypoxia/reoxygenation of micro-aerobic bags method. Then, the cytoplasm P65, P50, P-IκBα and nuclear P65, P50 content in olfactory ensheathing cells were detected by Western blot assay. Results Compared with normal serum control group, the cytoplasm and nucleus P65, P50, cytoplasm P-IκBαexpression in normal serum＋hypoxia group were significantly increased, the difference was statistically significant （P〈0.01）. Compared with normal serum control group, the cytoplasm and nucleus P65, P50, cytoplasm P-IκBα expression were lower in hypoxia＋edaravone group, hypoxia＋Jianpi Butu group, normal serum＋hypoxia＋PDTC group, hypoxia＋Jianpi Butu＋PDTC group, the difference was statistically significant （P〈0.05）. Conclusion Spleen-strengthening therapy can prevent the activation of NF-κB signal path, which may be due to its inhihitory effect on P65/P50 and P-IκBα overexpression in olfactory ensheathing cells with hypoxia/reoxygenation injury.