Neuroprotective effect of cerebroprotein hydrolysate on cerebral ischemia-reperfusion injury mice

OBJECTIVE To investigate the neuroprotective effect of cerebroprotein hydroly⁃sate(CH)on cerebral ischemia-reperfusion injury in mice.METHODS A total of 60 male SPF Kunming mice were randomly divided,reforming longa method into sham group(sham),model group(tMCAO,reforming longa method),CH 0.2 and 0.5 g·kg-1 groups and positive drug control group(edaravone 0.008 g·kg-1).Neurological deficit score were performed 24 h after opera⁃tion.Mice with scores ranged between 1 and 3 were included in subsequent experiments.Each group had 8 mice.CH edaravone and normal sa⁃line were ip injected for 5 d.The tMCAO group and the sham group were administered the same amount of normal saline as administration groups.TTC staining was used to measure the volume of cerebral infarction;ELISA was per⁃formed to detect the levels of interleukin-6(IL-6),interleukin-1β(IL-1β),brain-derived neurotrophic factor(BDNF)and interferon-γ(IFN-γ)in serum and penumbra.RESULTS TTC staining results showed that there was no infarction in sham group.Compared with tMCAO group,the infarct volume in each administration group was signifi⁃cantly decreased(P<0.01).ELISA results showed that IL-6,IL-1βand IFN-γin serum and penumbra were of significant difference between tMCAO group and sham group(P<0.01),and BDNF was significantly decreased(P<0.01).Compared with tMCAO group,IL-6,IL-1βand IFN-γin serum and ischemic penumbra were sig⁃nificantly decreased in all administration groups(P<0.01),while the content of BDNF was in⁃creased in CH 0.2 g·kg-1 group and edaravone 0.008 g·kg-1 group(P<0.05),and other groups were significantly increased(P<0.01).CONCLU⁃SION CH could reduce the cerebral infarction vol⁃ume and improve the nerve injury caused by cerebral ischemia-reperfusion.The mechanism was related to inhibit the expression of IL-6,IL-1βand IFN-γand increase the expression of BDNF possibly.

Establishment of lipopolysaccharide induced microglial inflammation model

OBJECTIVE To establish an in vitro inflammatory model of BV2 by observing the activity,the release amount of NO and the expression of inflammatory factors of microglial cells(BV2)induced by lipopolysaccharides(LPS).METHODS BV2 was routinely cultured in vitro.Cell viability was measured by CCK-8 meth⁃od.And by drew cell growth curve to determine the logarithmic growth cycle of the cells.After 24 h of routine culture,BV2 were induced by adding different concentrations of LPS(0.1,1.0 and 10.0 mg·L-1)for 4,8,12,24 and 48 h,respectively.Meanwhile,the morphological changes of BV2 were observed under inverted microscope to compare the activation degree of microglia at dif⁃ferent time and concentration.Cell activity and nitric oxide(NO)level were determined by CCK-8 and Griess method respectively,which could help to determine the optimal concentration and time of modeling.Finally,It were determined by ELISA that the concentrations of tumor necrosis factorα(TNF-α),interleukin-6(IL-6)and IL-1βin supernatant of LPS 1 mg·L-1 culture for 24 h.RESULTS BV2 were in logarithmic growth phase for 1 to 3 d after subculture.LPS 1 mg·L-1 induced BV2 for 24 or 48 h which could increase the release amount of NO significantly(P<0.05).In order to save time,LPS induced BV2 for 24 h were selected for subsequent experiments.Microglial cells in resting state were observed to be elongated spindle shape under inverted micro⁃scope.After LPS activation,the cell body became larger and the branching processes shrank back,presenting an amoeba-like appearance.ELISA results showed that the concentrations of TNF-α,IL-6 and IL-1βin supernatant of LPS 1 mg·L-1 cultured for 24 h were significantly increased which compared with the control group(P<0.05).CONCLUSION LPS could induce the activation of BV2 and up-regulate the level of inflammatory factors.The optimal condition for establishing stable BV2 microglial inflammatory model was used LPS 1 g·L-1 induced for 24 h.

Protective effect of hydroxysafflor yellow A on Parkinson cell model and its network pharmacology analysis

As an effective component of safflower,hydroxysafflor yellow A(HSYA)has the effect of promoting blood circulation,removing blood stasis,and relieving pain,and it has a certain effect on blood stasis and wind-induced Parkinson‘s disease(PD).However,the current research mostly involves a single intervention mechanism,which is not conducive to the clinical transformation of this type of drugs.In the present study,rotenone was used to construct a PD cell model,and the protective effect of HSYA on the PD cell model was evaluated by cell viability and mitochondrial membrane potential.TTD database was used to query PD-related therapeutic targets,Swiss Target Prediction database to query HSYA-related targets,STRING database was used to search the gene interaction relationship of common targets,and ClueGO pathway was adopted to enrich and analyze common targets and their interaction targets,as well as to explore the comprehensive intervention mechanism.The results showed that rotenone could successfully establish the PD cell model,and HSYA had significant protective effect on PD cell model.Through the network pharmacological analysis,36 PD-related therapeutic targets and 88 HSYA-related targets were queried.The common targets of PD and HSYA were FKBP1A,HTR1A,SLC6A4 and SLC6A3.To enrich four common targets and their interaction targets through REACTOME pathway,eight cell signal pathways were obtained,and six cell biological processes were obtained through biological process pathway enrichment.