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目的:观察宁夏无果枸杞叶提取物(FLE)对衰老小鼠脑海马区神经细胞增殖、凋亡及凋亡相关蛋白Bcl-2、Bax和Caspase-3表达的影响。方法:13月龄C57BL/6J雄性小鼠随机分为老年对照组和FLE低剂量组(FLE1组)、FLE中剂量组(FLE2组)、FIE高剂量组(FLE3组),6月龄为成年对照组。FLE1组、FIE2组、FLE3组分别用5 mg/kg、10 mg/kg、20 mg/kg FLE灌胃,0.2 mmol/I/(10 g·d),老年及成年对照组灌胃等量生理盐水,持续8周。流式细胞术检测海马区神经细胞凋亡率,免疫荧光组织化学方法观察小鼠海马齿状回神经细胞的增殖。免疫组化SP法及WesternBlot技术检测小鼠海马区凋亡蛋白Bcl-2、Bax、Caspase-3的表达,结果:流式细胞术检测结果显示,各组均检测到早期凋亡细胞,与成年对照组比较,老年对照组早期细胞凋亡率明显增多(P<0.05);FLE2组、FLE3组早期细胞凋亡率明显低于FLEl组和老年对照组(P<0.05)。免疫荧光染色结果显示:老年对照组海马齿状回神经细胞增殖率较成年对照组低(P<0.05),FLE2及FLE3组小鼠海马齿状回神经细胞增殖率较老年对照组增高(P<0.05)。免疫组化及Western Blot检测结果显示:老年对照组海马区Bcl-2蛋白的表达较成年对照组低(P<0.01);Bax、Caspase-3蛋白的表达较成年对照组高(P<0.01);FLE2及FLE3组Bcl-2蛋白的表达较老年对照组增加(P<0.01);Bax、Caspase-3蛋白的表达量较老年对照组降低(p<0.01)。结论:中、高浓度的FLE能够促进海马区神经细胞的增殖,提高凋亡蛋白Bcl-2表达,降低Bax、Caspase-3蛋白的表达,减少神经细胞的凋亡,发挥抗衰老作用。
OBJECTIVE: To observe the effects of Lycium barbarum leaf extract (FLE) on the proliferation, apoptosis and the expression of apoptosis-related proteins Bcl-2, Bax and Caspase-3 in the hippocampus of senile mice. Methods: Thirteen-month-old C57BL / 6J male mice were randomly divided into three groups: old control group, FLE1 low dose group, FLE2 middle dose group and FLE3 high dose group, Control group. FLE1 group, FIE2 group and FLE3 group were intragastrically administrated with 5 mg / kg, 10 mg / kg and 20 mg / kg FLE respectively, and 0.2 mmol / l (10 g · d) Brine for 8 weeks. Flow cytometry was used to detect the apoptosis rate of hippocampal neurons. Immunofluorescence histochemistry was used to observe the proliferation of dentate gyrus neurons in the hippocampus of mice. The expression of Bcl-2, Bax and Caspase-3 in the hippocampus of mice were detected by immunohistochemical SP method and WesternBlot technique. Results: Flow cytometry showed that early apoptotic cells were detected in all groups, Compared with the control group, the apoptosis rate of early control group was significantly increased (P <0.05). The early apoptosis rate of FLE2 group and FLE3 group was significantly lower than that of FLEl group and elderly control group (P <0.05). Immunofluorescence staining showed that the proliferation rate of hippocampal dentate gyrus neurons was lower in aged control group than in adult control group (P <0.05), and the proliferation rate of hippocampal dentate gyrus neurons in FLE2 and FLE3 groups was higher than that in old control group (P < 0.05). The results of immunohistochemistry and Western Blot showed that the expression of Bcl-2 protein in hippocampus of elderly control group was lower than that of adult control group (P <0.01), while the expression of Bax and Caspase-3 protein was higher than that of adult control group (P <0.01) ; The expression of Bcl-2 protein in FLE2 and FLE3 groups was higher than that in the old control group (P <0.01); The expression of Bax and Caspase-3 protein was lower than the old control group (p <0.01). CONCLUSION: FLE at medium and high concentration can promote the proliferation of neural cells in hippocampus, increase the expression of Bcl-2, decrease the expression of Bax and Caspase-3, decrease the apoptosis of nerve cells and exert anti-aging effects.