BACKGROUND:Hydroxy safflower yellow A has anti-ischemia,anti-oxidation,anti-thrombotic and anti-inflammatory effects.Whether it affects neuronal pyroptosis after glucose-oxygen deprivation/reglucose-reoxygenation is still unclear. OBJECTIVE:To investigate the protective effect of hydroxy safflower yellow A on neuronal pyroptosis and its mechanism. METHODS:HT22 cells in logarithmic growth phase were randomly divided into five groups:normal group,model group,hydroxy safflower yellow A group,colivelin group,and colivelin+hydroxy safflower yellow A group.HT22 cells were treated with glucose-oxygen deprivation/reglucose-reoxygenation to establish neuronal pyroptosis model,and then treated with STAT3 agonist Colivelin and hydroxy safflower yellow A.JC-1 probe was employed to assess changes in mitochondrial membrane potential.Reactive oxygen species kit was used to determine the content of reactive oxygen species in cells.GSDMD/TUNEL staining was conducted to observe cell pyroptosis.Immunofluorescence analysis was performed to detect STAT3 and GSDMD protein expression.RT-PCR was utilized for assessing mRNA expression levels of STAT3,NLRP3,and Caspase-1.Western blot assay was utilized to measure the protein expression levels of p-STAT3,NLRP3,GSDMD,Cleaved-caspase-1,and interleukin-1β. RESULTS AND CONCLUSION:(1)Compared with the normal group,the number of pyroptotic cells increased in HT22 cells in the model group along with a significant increase in protein expression levels of p-STAT3,NLRP3,Cleaved-caspase-1,GSDMD,and interleukin-1β.Compared with the model group,the number of pyroptotic cells reduced,and the expression of pyroptosis-related proteins significantly decreased in the hydroxy safflower yellow A group.(2)In comparison with the model group,pyroptosis worsened in the colivelin group where mitochondrial membrane potential decreased along with elevated reactive oxygen species content and increased mRNA expression levels of STAT3,NLRP3,and Caspase-1,as well as increased protein expression levels of p-STAT3,NLRP3,GSDMD,Cleaved-caspase-1,and interleukin-1β.Compared with the Colivelin group,above indexes were improved in the colivelin+hydroxy safflower yellow A group.These results suggest that hydroxy safflower yellow A plays a neuroprotective role through STAT3 signaling pathway to inhibit HT22 pyroptosis after glucose-oxygen deprivation/reglucose-reoxygenation treatment.

Objective To analyze the total alkaloids and six alkaloid components,including peimisine,of Fritillariae Anhuiensis Bulbus and five types of Fritillaria medicinal materials;To providebasis for the medicinal use of Fritillariae Anhuiensis Bulbus.Methods The total alkaloid content and 6 alkaloid components(peimisine,sipeimine-3β-D-glucoside,sipeimine,peimine,peiminine and isoverticine)of Fritillariae Anhuiensis Bulbus and 5 medicinal Fritillaria were determined by UV-Spectrophotometry and UPLC-ELSD method,and 18 batches of Fritillaria medicinal materials were classified by clustering analysis and principal component analysis.Results The total alkaloid content of Fritillariae Anhuiensis Bulbus was significantly higher than that of other Fritillarias.The results of clustering and principal component analysis showed that Fritillariae Anhuiensis Bulbus was classified into one class with Fritillariae Thunbergii Bulbus and Fritillariae Hupehensis Bulbus.Conclusion Fritillariae Anhuiensis Bulbus is similar to Fritillariae Thunbergii Bulbus and Fritillariae Hupehensis Bulbus in the composition of alkaloids,which can be used as a medicinal cultivation variety.

Objective To investigate the effects of berberine combined with quercetin on the rats with polycystic ovary syndrome(PCOS)based on toll-like recepter 4(TLR4)/nuclear-factor-KB(NF-κB)signaling pathway.Methods Thirty SD female rats were stratified by weight and then randomly divided into the blank control group,model control group,berberine group,quercetin group and berberine+quercetin group,6 rats in each group.The rats conducted daily subcutaneous injection of dehydroepiandrosterone(DHEA)for con-structing the PCOS model.After consecutive medication intervention for 21 d,the body weight and ovary mass of the rats in each group were weighed,and the hematoxylin-eosin(HE)staining was used for conducting the histological morphological observation on ovarian tissues.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of serum luteinizing hormone(LH),follicle stimulating hormone(FSH),testosterone(T),interleukin-6(IL-6)and tumor cytokine alpha(TNF-α).The Western blot was used to determine the relative levels of TLR4 and NF-κB protein.Results Compared with the blank control group,the ovaries in the model control group were manifested by polycystoid,atretic follicles increase,body weight and ovarian mass increase(P＜0.01),the increase of LH,T,IL-6 and TNF-α levels(P＜0.01),decrease of FSH level(P＜0.01)and increase of TLR4 and p-NF-κBp65 protein expression levels(P＜0.01).Compared with the model control group,the ovarian cystic dilatation in the berberine group,quercetin group and the berberine+querce-tin group was decreased,the atretic follicles were decreased,the local ovarian morphology was similar to that of the blank control group.Compared with the model control group,the body weight in the berberine group was decreased(P＜0.05),the levels of LH,T,IL-6 and TNF-α in the berberine group and quercetin group were decreased(P＜0.01),the levels of TLR4 and p-NF-κBp65 protein expression were decreased(P＜0.01)and the ovarian mass was reduced(P＜0.01).But the FSH and the body weight level had no statistical differ-ence between the berberine group and quercetin group(P＞0.05).The body mass and ovarian mass in the berberine+quercetin group were decreased(P＜0.01),the FSH level was increased(P＜0.01),the LH,T,IL-6 and TNF-α levels were decreased(P＜0.01),and the TLR4 and p-NF-κBp65 protein expression levels were decreased(P＜0.01).Conclusion Berberine combined with quercetin may regulate the FSH,LH and T hormone secretion,decrease the release of IL-6 and TNF-α inflammatory factors and improve the ovarian func-tion possibly by inhibiting the TLR4/NF-κB signaling pathway.