目的:探讨补阳还五汤含药血清对体外缺氧损伤神经干细胞(NSCs)增殖及Notch信号通路相关因子m RNA表达的影响。方法:体外分离培养小鼠胚胎NSCs,取第3代悬浮生长的NSCs随机分组,空白组予以无血清条件培养基常氧培养,其余3组分别给予缺氧4h、缺氧4h加10%正常血清和缺氧4h加10%补阳还五汤含药血清干预;培养第5天观察细胞增殖情况,同时提取细胞总RNA,Real-time PCR检测Notch通路4个关键节点因子Notch-1、Hes-1、Neurogenin-1(Ngn-1)和Delta-like-1(Dll-1)m RNA相对表达水平。结果:正常生长的NSCs Notch-1、Hes-1 m RNA高表达,Ngn-1、Dll-1 m RNA低表达;缺氧4h后NSCs克隆球数减少(P<0.05),Notch-1、Hes-1 m RNA下调(P<0.05),Ngn-1、Dll-1 m RNA上调(P<0.05);10%补阳还五汤含药血清能显著上调缺氧NSCs Notch-1、Hes-1、Ngn-1 m RNA以及下调Dll-1 m RNA的表达水平,增加缺氧NSCs克隆球数,与10%正常血清相比差异显著(P<0.05)。结论:调节NSCs Notch信号通路相关因子Notch-1、Hes-1、Ngn-1、Dll-1 m RNA的表达水平可能是补阳还五汤促进缺氧损伤后NSCs增殖的机制。
目的观察补阳还五汤对Cav-1^-/-小鼠脑缺血模型PI3K、Akt、PTEN表达的影响,探讨其可能的作用机制。方法将野生型小鼠(WT)及Cav-1^-/-小鼠(KO)随机分为假手术组、模型组与补阳组。采用大脑中动脉栓塞法制备脑缺血模型,干预10 d后运用Ayelet Levy 14分评分法观察小鼠神经功能评分。免疫组化和qRT-PCR分别检测小鼠脑组织PI3K、Akt、PTEN蛋白和mRNA的表达。结果与假手术组比较,各模型组小鼠均出现明显神经功能障碍(P<0.01),且PI3K、Akt蛋白和mRNA表达明显升高(P<0.01),PTEN蛋白和mRNA表达明显降低(P<0.01),WT模型组小鼠脑组织PI3K、Akt、PTEN蛋白和mRNA表达更显著,且神经功能评分优于KO模型组(P<0.01,P<0.05);WT补阳组小鼠脑组织PI3K、Akt、PTEN蛋白和mRNA的调控程度及神经功能恢复程度均优于WT模型组和KO补阳组(P<0.01)。结论补阳还五汤可能通过Cav-1调控PI3K/Akt信号通路活性,发挥抗脑缺血损伤的作用。
Objective:To investigate the effect of Buyang Huanwu Decoction(补阳还五汤,BYHWD) on estradiol(E_2) and estradiol receptor(ER) in serum and brain in ovariectomized rats after middle cerebral artery occlusion(MCAO).Methods:Adult female rats were ovariectomized and focal cerebral ischemic was induced by MCAO.Rats were randomly divided into normal,ovariectomy(OVX),MCAO,OVX+MCAO,OVX+MCAO+E_2,and OVX+MCAO+BYHWD group.Rats were administered BYHWD 5 g/kg daily,estradiol valerate 500 μg/kg per day or distilled water for 7 consecutive days.Neuronal function and infarct volume were measured on day 7 after artery occlusion,and E_2 and ER concentration in serum and brain were checked by enzyme-linked immunosorbent assay.Results:BYHWD significantly improved the neurological behavior,reduced the infarction volume,increased E_2concentration in serum and brain,and increased ER concentration in the brain in ovariectomized rats after MCAO.Conclusion:The neuroprotective effects of BYHWD are associated with estrogen and its receptor.
