Objective:To investigate the effects of Apelin-13 regulatory peptide on neuronal cell pyroptosis in mice modeled with cerebral ischemia-reperfusion(I/R).Methods:We prepared a mouse cerebral I/R model using middle cerebral artery embolization and Reperfusion(MCAO/R).The HT22 cell injury model was prepared by the oxygen glu-cose deprivation/reoxygenation(OGD/R),and Apelin-13 treatment was also given.Neurological function was assessed by neurological deficit score;hematoxylin-eosin(HE)staining and Nissl staining were used to observe the morphologic changes of the infarcted area of the mice;and 2,3,5triphenyltetrazolium chloride(TTC)staining was used to observe the volume of cerebral infarcts;The expression of NOD-like receptor thermoprotein structural domain-related protein 3(NLRP3),gasdermin D(GSDMD),caspase-1,apoptosis-associated speck-like protein(ASC),interleukin 1β(IL-1β),and interleukin 18(IL-18)in brain tissues from infarcted areas or HT22 cells was detected by Western Blot,and IL-1β and IL-18 were detected by enzyme-linked immunosorbent assay(ELISA)in serum of mice and culture supema-tants;The cell viability and cell damage of HT22 were detected by CCK-8 kit and lactate dehydrogenase(LDH)assay kit,respectively;caspase-1 activity was measured by caspase-1 activity kit in HT22 cells;and the expression of caspase-1 and GSDMD was observed by immunofluorescence staining in HT22 cells.Results:Apelin-13 significantly improved neurological function and cerebral infarct volume in I/R mice,and attenuated pathological damage in the in-farcted area.It also reduced the serum levels of IL-1β and IL-18.In addition,Apelin-13 reduced the expression of mol-ecules such as NLRP3,GSDMD,caspase-1,IL-1β,and IL-18 in the cerebral infarct area of mice.In vitro experiments showed that Apelin-13 significantly increased the viability of OGD/R-treated HT22 cells,decreased caspase-1 activity,and reduced the LDH content,as well as decreased the expression of molecules such as NLRP3,GSDMD,caspase-1,IL-1β,IL-18,and so on,in OGD/R-treated HT22 cells.Conclusion:Apelin-13 inhibits pyroptosis through the NL-RP3/caspase-1/GSDMD pathway in cerebral ischemia/reperfusion mice and thus exerts neuroprotective effects.

Objective:To investigate whether Lycium barbarum polysaccharides(LBP)can inhibit ferroptosis through the solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)pathway and thus alleviate cerebral ischemia-reperfusion(I/R)injury.Methods:The mouse model of cerebral I/R injury was established by mid-dle cerebral artery occlusion(MCAO)and treated with Lycium barbarum polysaccharides(LBP).The mice were ran-domly divided into three groups:sham group,I/R group,and I/R+LBP group.2,3,5triphenyltetrazolium chloride(TTC)staining was used to observe the volume of cerebral infarcts;Neurological function was assessed by neurological deficit score;The superoxide dismutase(SOD),glutathione(GSH),malondialdehyde(MDA),and Fe2+detection kit to detect the changing levels of SOD,GSH,MDA,and Fe2+;Western Blot detects the expression of ferroptosis-related keys GPX4,SLC7A11,and Nrf2.Results:LBP can reduce the cerebral infarction volume and improve the neurological function.LBP increases the content of SOD and GSH and reduces the content of MDA in I/R injured mice.Western Blot results show that the expression levels of ferroptosis-related proteins GPX4,SLC7A11,and Nrf2 in the brain tissue of the I/R group were significantly lower than those of the Sham group.After LBP treatment,the levels of GPX4,SLC7A11,and Nrf2 in the LBP+I/R group were significantly higher than those of the I/R group.Conclusion:LBP at-tenuates cerebral I/R injury by inhibiting ferroptosis through the SLC7A11/GPX4 pathway.LBP has a neuroprotective effect on cerebral I/R injury and is a potential neuroprotective agent.

Objective:To investigate whether Lycium barbarum polysaccharides(LBP)can inhibit ferroptosis through the solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)pathway and thus alleviate cerebral ischemia-reperfusion(I/R)injury.Methods:The mouse model of cerebral I/R injury was established by mid-dle cerebral artery occlusion(MCAO)and treated with Lycium barbarum polysaccharides(LBP).The mice were ran-domly divided into three groups:sham group,I/R group,and I/R+LBP group.2,3,5triphenyltetrazolium chloride(TTC)staining was used to observe the volume of cerebral infarcts;Neurological function was assessed by neurological deficit score;The superoxide dismutase(SOD),glutathione(GSH),malondialdehyde(MDA),and Fe2+detection kit to detect the changing levels of SOD,GSH,MDA,and Fe2+;Western Blot detects the expression of ferroptosis-related keys GPX4,SLC7A11,and Nrf2.Results:LBP can reduce the cerebral infarction volume and improve the neurological function.LBP increases the content of SOD and GSH and reduces the content of MDA in I/R injured mice.Western Blot results show that the expression levels of ferroptosis-related proteins GPX4,SLC7A11,and Nrf2 in the brain tissue of the I/R group were significantly lower than those of the Sham group.After LBP treatment,the levels of GPX4,SLC7A11,and Nrf2 in the LBP+I/R group were significantly higher than those of the I/R group.Conclusion:LBP at-tenuates cerebral I/R injury by inhibiting ferroptosis through the SLC7A11/GPX4 pathway.LBP has a neuroprotective effect on cerebral I/R injury and is a potential neuroprotective agent.

Objective To observe the effect of salted processing on the contents of total alismol,24-acetylalisol A and 23-acetylalisol B in A lisma Rhizoma.Methods UV was used for the determination of total alismol′s content in crude A lisma Rhizoma and salted A lisma Rhizoma with Meta Nitrobenzene and Potassium Hydroxide as the developers.The detection wave-length was 545 nm.HPLC was used for the determination of 24-acetylalisol A′s and 23-acetylalisol B′s contents in crude A lis-ma Rhizoma and salted A lisma Rhizoma with Elite C18(4.6 mm×250 mm,5 μm)as the chromatographic column.The mobile phase was gradient elution with acetonitrile and water.The detection wavelength was 208 nm.The column temperature was set at 25℃.The flow rate was 1 ml/min.Results The linear relationships of total alismol,24-acetylalisol A and 23-acetylalisol B were fine(r>0.999 0)in the linear ranges by UV and HPLC.The average recoveries of them in crude A lisma Rhizoma and salted A lisma Rhizoma ranged from 97.81%-100.05%,with the RSD less than 3%.The average contents of total alismol,24-acetylalisol A and 23-acetylalisol B in ten batches of crude A lisma Rhizoma were 4.41%,0.208%,0.065% respectively.The average contents of them in ten batches of salted A lisma Rhizoma were 9.62%,0.232%,0.017% respectively.Conclusion The contents of total alismol,24-acetylalisol A and 23-acetylalisol B in A lisma Rhizoma were influenced considerably by salted processing.The content of total alismol increased obviously.The content of 24-acetylalisol A increased slightly.The content of 23-acetylalisol B decreased obviously.