Objective To investigate the effect of gene of phosphate and tension homology deleted on chromsome ten (PTEN) inhibitor bpV (pic) on the proliferation and differentiation of neural stem cells (NSCs).Metods Stem cell suspension was prepared from the cerebral cortex of the 16 days of fetal rat.The cells were divided into2 groups:group A (control group):no treatment,group B (experimental group):bpV treatment group (culture medium containing bpV).The number of stem cell spheres was observed by immunofluorescence staining,the proliferation of NSCs was detected by cell counting kit-8 (CCK-8)test,the differentiation of NSCs was detected by immunofluorescence staining,and the expression of PTEN and the mammalian target of rapamycin (mTOR) genes was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) on the fifth day.Results In the experimental group,the number of stem cells per low field (× 50) in the experimental group was 60 ± 5,which was significantly higher than that in the control group (40 ± 3,P ＜ 0.01).The CCK-8 test confirmed that the absorbance value of the experimental group at 425 nm was 0.997 ± 0.085,which was higher than that of the control group 0.788 ± 0.083 (P ＜0.01).The percentage of NSCs differentiated into neurons in the experimental group was (27.860 ±1.927)％,which was significantly higher than that in the control group [(13.120 ± 1.130)％,P＜0.01],while the percentage of NSCs differentiated into glia was (61.900 ± 1.840) ％,which was significantly lower than that in the control group [(77.520 ± 1.035) ％,P ＜ 0.01];the percentage of NSCs differentiated into oligodendrocytes was (5.320 ± 0.975)％ in the experimental group and (4.460 ± 0.586) ％ in the control group (P ＜ 0.05);the percentage of NSCs differentiated into other cells was (4.780 ± 0.572) ％ in the experimental group and (4.900 ± 1.208) ％ in the control group (P ＞ 0.05).The expression of PTEN in experimental NSCs was 2.13 times higher than that in control NSCs (P ＜0.01),while the expression of mTOR in experimental NSCs was 3.62 times higher than that in control NSCs (P ＜ 0.01).Conclusion PTEN inhibitor bpV (pic) can promote the proliferation of neural stem cells and promote the differentiation of neural stem cells into neurons to a certain extent.%目的 观察第10号染色体上缺失与张力蛋白同源的磷酸酯酶基因(PTEN)抑制剂牛细小病毒bpV(pic)对神经干细胞(NSCs)增殖及分化的影响.方法 取胚胎16 d SD大鼠的大脑皮层,制成干细胞悬液,细胞分组为:A组(对照组):不做任何处理;B组(实验组):bpV处理组(培养液中含bpV).于第5天用免疫荧光染色观察两组干细胞球的数量,行细胞计数试剂盒(CCK-8)试验检测NSCs增殖,免疫荧光染色检测NSCs分化,反转录-聚合酶链反应(RT-PCR)检测两组细胞在第5天时PTEN和哺乳动物雷帕霉素靶蛋白(mTOR)基因的表达.结果 实验组的每个低倍视野(×50)干细胞数量为60±5,明显多于对照组(40±3,P＜0.01).CCK-8试验证实实验组在425nm处的吸光度值为0.997±0.085,高于对照组(0.788±0.083,P＜0.01).实验组NSCs分化为神经元的百分率为(27.860±1.927)％,明显多于对照组[(13.120±1.130)％,P＜0.01];而分化为胶质细胞的百分率为(61.900±1.840)％,明显少于对照组[(77.520±1.035)％,P＜0.01];分化为少突胶质细胞的百分率实验组为(5.320±0.975)％,对照组为(4.460±0.586)％ (P ＜0.05);分化为其他细胞的百分率实验组为(4.780±0.572)％,对照组为(4.900±1.208)％ (P ＞0.05).实验组NSCs中PTEN的表达量是照组NSCs中PTEN表达量的2.13倍(P＜0.01),而实验组NSCs中mTOR的表达量是照组NSCs中mTOR表达量的3.62倍(P＜0.01).结论 PTEN抑制剂bpV(pic)可促进神经干细胞增殖,并在一定程度上促进神经干细胞向神经元分化.
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