Effect Of Periplaneta Americana Extract On Nerve Cell Proliferation And Apoptosis

Purpose:In vivo experiments were conducted to study the effects of Kangfuxin fluid,an adult drug extracted from Periplanella americana,on the proliferation of neurons in the dentate gyrus hippocampus of C57BL/6 mice and 3-transgenic Alzheimer’s disease model mice,as well as the learning and memory ability of the mice.The mechanism of Kangfuxin fluid to improve the learning and memory ability of mice was also discussed.The effects of different components of Periplaneta americana extracts on the proliferation and apoptosis of mouse hippocampal neuron cell lines were studied in vitro.Finally,the effect of Periplaneta americana extract on learning and memory and its possible mechanism were elucidated,providing a basis for the use of Periplaneta americana extract to improve cognitive dysfunction caused by neural aging.Method:1.Twenty 4-month-old healthy C57BL/6 male mice were randomly divided into control group and administration group.After 8 weeks of administration,5-ethynyl-2’-deoxyuridine(Ed U)was injected intraperitoneally.After 4 days of perfusion,the brain was taken and the number of new neurons in the dentate gyrus of the hippocampus of mice was detected by Ed U staining.2.Twenty 6-month-old 3-transgenic AD model mice were randomly divided into control group and administration group.After 8 weeks of treatment,Ed U was injected intraperitoneally.The learning and memory ability of AD model mice was detected by Y maze method.After 4 days of cerebral perfusion,the expression levels of Ed U,β-catenin and GSK-3β in the hippocampus dentate gyrus were detected.3.The mouse hippocampal neuronal cell line(HT22 cells)was cultured in vitro,and HT22 cells were treated with water extract,40% ethanol extract,60% ethanol extract,95% ethanol extract and crude extract of Periplanata americana,respectively.Cell proliferation and cell apoptosis were detected by cell counting and CCK-8,and cell apoptosis was detected by flow cytometry.Results:1.The number of new neurons in hippocampal dentate gyrus of C57BL/6 mice in Kangfuxin fluid administration group was significantly higher than that in control group(P<0.01).2.In Y maze test,the training times of reaching the target(P<0.05)and the error times of reaching the target(P<0.05)in the 3-transgenic AD model mice in Kangfuxin fluid administration group were significantly higher than those in the control group;The number of errors in memory ability test was significantly lower than that in control group(P<0.05).Compared with the control group,the Ed U staining and the number ofβ-catenin immunopositive cells in the hippocampal dentate gyrus of the 3-transgenic AD model mice in the Kangfuxin fluid administration group were significantly increased(P<0.05),and the number of GSK-3β immunopositive cells were significantly decreased(P<0.05).3.The cell count and CCK-8 assay of different components of Periplaneta americana extracts showed that water extract and 40% ethanol extract of Periplaneta americana significantly promoted cell proliferation(P<0.05);The 60% ethanol extract of Periplaneta americana had no effect on cell proliferation.95% ethanol extract and crude extract of Periplaneta americana inhibited cell proliferation(P<0.05).4.Flow cytometry showed that water extract and 40% ethanol extract of Periplaneta americana inhibited HT22 cell apoptosis compared with solvent control group(P<0.01);95% ethanol extract and crude extract of Periplaneta americana promoted HT22 cell apoptosis(P<0.01).The 60% ethanol extract of Periplaneta americana showed no significant difference in apoptosis of HT22 cells(P>0.05);Apoptosis rate of water extract of Periplaneta americana was the lowest at the same concentration.Conclusion:1.Kangfuxin liquid can improve the learning and memory ability of 3-transgenic AD model mice.2.Kangfuxin fluid can promote the proliferation of neurons in the hippocampal dentate gyrus of C57BL/6 mice and 3-transgenic AD model mice.3.Kangfuxin fluid may promote the regeneration of hippocampal neurons by up-regulating the expression of β-catenin in the dentate gyrus of 3-transgenic AD model mice,down-regulating the expression of GSK-3β,and activating the Wnt/β-catenin signaling pathway,thus improving the learning and memory ability of AD model mice.4.The main components of Periplaneta americana extract to improve learning and memory may exist in water extract and 40% ethanol extract of Periplaneta americana.