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（E2） 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＋E2,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 E2 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 E2concentration 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.
