| Objective:To study the effect of epimedium wushanense flavonoids on depression-like behavior of prenatal stress offspring male rats by observing their behavioral changes.To explore the the improvement effect and mechanism of epimedium wushanense flavonoids on depression-like behavior,a series of experiments had been carried out by detecting the changes of the content of malondialdehyde,superoxide dismutase activity and catalase activity in serum,the concentration of corticosterone in serum,and the expression of brain-derived neurotrophic factor BDNF and its receptor TrkB in hippocampus.Methods:1.The SD pregnancy rats were used to establish the model of prenatal chronic restraint stress.The pregnant rats were randomly allocated to 5 groups(6 per group):①Controlgroup(C),②prenatal stress group(PS),③Prenatal stress+saline group(NS),④Prenatal stress+EWFD group(EWFD,500 mg/kg/day),⑤Prenatal stress+EWFZ group(EWFZ,300 mg/kg/day).On the 7th to 21st day of pregnancy,the PS+NS group was given normal saline once a day,and the stress groups(PS+EWFD,PS+EWFZ)were given EWF daily for 14 days,and each stress group was given restraint stress,the control group was fed normally without any intervention.The offspring rats were randomly selected 1 to 2 male rats and divided into OC,OPS,ONS,OEWFD and OEWFZ groups(6 per group)according to the corresponding groups of the pregnant rats.All the offspring rats were sampled to behavioral experiments on the 30th day after birth,then they were subjected to having hippocampus decapitated,blood taked from abdominal aorta,serum separated,and various indexes measured.2.The effects of epimedium wushanense flavone on the depression-like behaviors of prenatal stressed offspring male rats were evaluated with experiment of sugar water preference,forced swimming and open field.3.The effects of epimedium wushanense flavone on CORT concentration in serum of PS offspring rats were observed by enzyme-linked immunosorbent assay(ELISA)Kit.4.The effects of epimedium wushanense flavone on oxidative stress marker were observed by detecting changes of the content of MDA and the activity of SOD and CAT in serum of PS offspring rats.5.The effects of epimedium wushanense flavone on the protein expression of BDNF and TrkB in hippocampus of PS male offspring rats were observed by Western blot.6.The effects of epimedium wushanense flavone on BDNF and TrkB mRNA expressions in the hippocampus of PS male offspring rats were observed by real-time quantitative PCR.Results:1.The results of sugar water preference experiment showed that the sugar water consumption of OPS group was significantly lo wer than OC group(p<0.01).Compared with ONS group,the preference of sugar water in OEWFD and OEWFZ was significantly increased(p<0.01).There were significant difference in total drinking water between the other groups of rats and blank control group(p<0.01).2.The results of force-swimming experiment of PS induced male offspring rats showed that the immobility time was significantly prolonged which compared with OC group(p<0.01).Compared with ONS group,OEWFD and OEWFZ group significantly reduced the immobility time of prenatal stress offspringrats in force-swimming(p<0.01,p<0.05),and the high dose group reduced the immobility time of prenatal stress offspring rats more than the low dose group.3.The results showed that the number of horizontal crossing cells in OPS group was significantly lower than that in OC group(p<0.05),the residence time of central cell was significantly longer(p<0.01),the number of erections was significantly lower(p<0.01),and the number of fecal granules was significantly lower(p<0.01).Compared with ONS group,the number of crossing cells of prenatal stress rats increased significantly(p<0.05),the residence time in the central cell decreased significantly(p<0.01),the number of erections increased significantly(p<0.05),the number of modified cells increased significantly(p<0.01),and the number of fecal particles increased significantly(p<0.01)of OEWFD group.4.The results showed that serum corticosterone concentration of PS male offspring rats were increased(p<0.01),the serum corticosterone concentration were decreased by EWF compared with the PS male offspring rats group(p<0.01),significantly.The results showed that EWF could improve depressive-like symptoms of PS male offspring rats by regulating the concentration of corticosterone.5.The determination of antioxidant index showed that the total SOD activity and CAT activity in serum of male rats were significantly decreased(p<0.01),MDA content was significantly increased(p<0.01)of PS offspring rats,which reflected that the body’s ability to scavenge oxygen free radicals was weakened,the ability to protect cells from damage was reduced.Compared with ONS group,the total SOD activity and CAT activity of OEWFD and OEWFZ were strengthened by epimedium wushanense flavone(p<0.01),MDA content became much lower(p<0.01),which reflected that the scavenging capacity of oxygen free radicals were enhanced by EWF intervention6.The results of western-blot showed that the protein expressions of BDNF and TrkB in hippocampus of PS male offspring rats were significantly lower than OC group(p<0.01).Compared with ONS group,the protein expressions of BDNF in hippocampus were significantly increased with the intervention of EWFD(p<0.01).The protein expressions level of TrkB were significantly increased with EWFD or EWFZ(p<0.01).7.The results of real-time quantitative PCR showed that the mRNA expression of BDNF and TrkB were significantly decreased in hippocampus of PS male offspring rats(p<0.01).Compared with ONS group,the expression of BDNF and TrkB mRNA in hippocampus of PS male offspring rats were increased by EWFD and EWFZ(p<0.01),significantly.Conclusion:1.EWF could improve depression-like behavior of male offspring rats caused by PS.2.The increase of CORT concentration could induced depression-like behavior of PS male offspring rats.EWF has improved the depression-like behavior of PS rats by inhibiting this change.3.EWF has changed the activity decrease of SOD and CAT,and the content increase of MDA in serum of male offspring rats caused by PS.Furthermore,it has improved the depression-like behavior of PS male offspring rats.4.EWF has changed the down-regulation of protein expression of BDNF and TrkB in hippocampus of male offspring rats caused by PS.Moreover,it has improved the depressive-like behavior of PS male offspring rats via up-regulating their protein expression level.5.EWF has changed the down-regulation of mRNA expression of BDNF and TrkB in hippocampus of male offspring rats caused by PS.Furthermore,it has improved the depressive-like behavior of PS male offspring rats via up-regulating their mRNA expression level. |
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