壮通饮对体外培养新生大鼠海马神经干细胞增殖分化的影响

Effect of “Zhuang Tong Yin”on Proliferation and Differentiation of Hippocampus Neural Stem Cell of Cultured Neonatal Rat

探讨壮通饮对体外培养大鼠大脑海马区内源性神经干细胞(neural stem cells,NSCS)分化作用的影响。利用无血清DMEM/F12(含20 ng/mL bFGF和EGF及2%B27)体外培养技术从新生Wistar大鼠大脑海马区中培养NSCS,在NSCS分化过程中添加不同剂量(20、40和80 mg/L)壮通饮进行干预,以免疫荧光及细胞化学方法对NSCS及其分化后的细胞进行鉴定。从大脑海马区分离培养的NSCS能够表达巢蛋白(Nestin),并具有分化为神经元、星形胶质细胞及少突胶质细胞的能力。在同一接种密度条件下,不同剂量壮通饮干预组的Nestin阳性细胞表达数分别为:20 mg/L组:21.5±2.9;40 mg/L组:18.6±2.4;80 mg/L组:16.7±2.2,与对照组(24.8±3.1)比较明显减少(P<0.05)。而壮通饮干预组表达神经元核心抗原(NeuN)、胶原纤维酸性蛋白(GFAP)、髓鞘碱性蛋白(MBP)的阳性细胞数较对照组明显增加(P<0.05)。通过本实验证实壮通饮能诱导大脑海马区NSCS分化,具有一定的促进神经再生的作用。

The objective of this study was to explore the effect of a Chinese proprietary medicine ＂Zhuang Tong Yin＂ on proliferation and differentiation of cultured rat brain hippocampus region derived neural stem cells （ neural stem cells, NSCS）. Serum2free DMEM/F12 （containing 20 ng/mL of bFGF, EGF and 2% B27） in vitro culturing technique was used to culture hippocampus NSCS from newborn Wistar rats. In the NSC$ differentiation process, different doses （20 mg/L,40 mg/L and 80 mg/L） of ＂Zhuang Tong Yin＂ were added to intervene. Immunottuorescence and cytochemieal methods were used to identify NSCS and its differentiated cells. The results showed that NSCS isolated and cultured from the hippocampus region of the brain was able to express Nastin and it can be differentiate into neurons, astrocytes and ol- igedendrocytes. Under the same inoculation density condition, the expressions of Nestin-positive cells, of different dose groups of ＂Zhuang Tong Yin＂ were：20 mg/L group：21.5 ~ 2.9;40 mg/L group：IS.6 ~ 2.4;80 mg/L group：16.7± 2.2, decreased significantly （P 〈 0. 05） compare with with control group （24.8± 3.1 ）. In addition, the expression of neurons core antigen （NeuN） ,glial fibrillary acidic protein （GFAP） ,myelin basic protein （MBP） cells significantly increased compared with the control group （P 〈 0.05）. This experiment confirmed that ＂Zhuang Tong Yin＂ can induce the brain hippocampus NSCS differentiation, has a certain role in promoting nerve regeneration.