加味五子衍宗方含药脑脊液对β淀粉样蛋白诱导海马神经元损伤的保护作用

Protective Effect of Cerebrospinal Fluid Containing Jiawei Wuzi Yanzong Formula on β-amyloid Proteininduced Injury of Hippocampal Neurons

目的研究加味五子衍宗方及其总多糖、总黄酮含药脑脊液对β淀粉样蛋白诱导的原代大鼠海马神经元损伤的保护作用。方法体外原代培养大鼠胎鼠海马神经元,并将加味五子衍宗方的提取物(总方、总多糖、总黄酮)对大鼠进行灌胃给药,然后抽取含药脑脊液,将含药脑脊液与20μmol/L的Aβ25-35共同作用海马神经元24h后,分别检测正常对照组、Aβ25-35损伤组、空白脑脊液保护组、加味五子衍宗方含药脑脊液(总方、总多糖、总黄酮)保护组的海马神经元活力、凋亡率以及凋亡相关蛋白(caspase-3,PARP,Bcl-2,Bcl-XL,JNK1/2,p38MAPK)的变化情况。结果加味五子衍宗方不同提取物(总方、总多糖、总黄酮)的含药脑脊液对Aβ25-35所诱导的海马神经元毒性均有保护作用,且总黄酮、总多糖含药脑脊液的保护作用明显高于总方含药脑脊液(P<0.05或P<0.01),但是加味五子衍宗方不同组分(总方、总多糖、总黄酮)的含药脑脊液对凋亡相关蛋白的调控机制并不一致。结论加味五子衍宗方中的部分黄酮类和多糖类成分可以透过血脑屏障并进入脑脊液,同时这些成分对β淀粉样蛋白所致的神经损伤有一定的保护作用。

Objective To investigate the protective effects of cerebrospinal fluid（CSF）-containing Jiawei Wuzi Yanzong formula（CSF-C） on β-amyloid protein-induced injury of hippocampal neurons.Methods Primary hippocampal neurons were isolated and cultured for 7 days in vitro.CSF-C was prepared by cerebrospinal fluid extracted from SD rats fed continuously with various components of Jiawei Wuzi Yanzong formula（total formula,total flavonoids or total polysaccharides） respectively.The viability,morphological change,apoptotic rate and apoptosis-related proteins（caspase-3,PARP,Bcl-2,Bcl-XL,JNK1 /2 and p38 MAPK） of neurons were detected in different groups：the untreated normal group,the Aβ25-35-treated group,and the CSF-C protected groups（coacted by CSF-C and 20 μmol/L of Aβ25-35 ）,respectively.Results CSF-C showed significant neuro-protective effect,and the protection of CSF-C contained total flavonoids or total polysaccharides was significantly greater than that contained total formula（P 0.05 or P 0.01） .Moreover,the effects of CSF-C contained various components on apoptosis-related proteins were different.Conclusion Some flavonoid and polysaccharide components in Jiawei Wuzi Yanzong formula can pass through the blood brain barrier and protect neurons from β-amyloid protein-induced neuron injury to some extents.