竹荪多糖通过抑制NLRP3/GSDMD/IL-1β信号通路缓解慢性乙醇暴露小鼠小脑浦肯野细胞的损伤OA

Objective:To explore the alleviating effects of dictyophora polysaccharide(DIP)on the damage of Purkinje cells(PC)in the cerebellum of mice caused by chronic ethanol exposure through inhibiting the NLRP3/GSDMD/IL-1β signaling path-way.Methods:A mouse model of chronic ethanol exposure was constructed.The mice were divided into 5 groups(n=8)according to the random number table method:the Control(Con)group,the 100 mg/kg DIP(DIPH)group,the 4 g/kg 20%ethanol(EtOH)group,the 4 g/kg 20%ethanol+25 mg/kg DIP(EtOH+DIPL)group,and the 4 g/kg 20%ethanol+100 mg/kg DIP(EtOH+DIPH)group.The gait test and rotarod test were used to detect the motor coordination and balance ability of the mice,the expres-sion levels of inflammatory factors interleukin-1β(IL-1β)and interleukin-18(IL-18)were detected by ELISA.Hematoxylin-eosin staining(H&E)was used to observe the morphology of PC.Nissl staining was used to observe the distribution and morphology of Nissl bodies in PC.Calbindin staining was used to observe the morphology,distribution,and number of PC.TUNEL staining was used to observe the apoptosis of PC.Glial fibrillary acidic protein(GFAP)staining and ionized calcium-binding adapter molecule-1(Iba-1)staining were used to observe the proliferation state of glial cells.Staining with 2',3'-cyclic-nucleotide 3'-phosphodiesterase(CNP)was used to reflect the integrity of myelin sheaths.Gomori's silver impregnation staining was used to observe the changes of axons of PC.Transmission electron microscopy(TEM)was used to observe the surface morphology of PC,the number of syn-apses,and the damage of cell membranes,cell nuclei,and mitochondria.Western blot was used to detect the expression levels of proteins related to the NLRP3 inflammatory pathway in cerebellar tissues.Golgi staining was used to observe the changes of den-dritic spines of PC.Results:Compared to the Con group,the motor coordination and balance ability of mice in the EtOH group were significantly decreased(P＜0.01).H&E staining,Nissl staining,and Calbindin staining showed that in the EtOH group,the number of PC was significantly decreased(P＜0.01),the number of GFAP and Iba-1 positive cells was significantly increased(P＜0.01),and a large number of PC had demyelinating lesions,as well as the loss of axon trees and dendrites(P＜0.01).The ex-pression levels of downstream proteins of the NLRP3 inflammatory pathway were significantly increased(P＜0.01).TEM showed that the synaptic density was significantly decreased(P＜0.01),the length of the postsynaptic density(PSD)became shorter and the thickness became thinner(P＜0.01),cristae structure of mitochondria in neurons disappeared,and the mitochondi-ra was vacuolized.After DIP intervention,the motor coordination and balance ability significantly improved,the number and mor-phology of PC became normal,the expression levels of downstream proteins of the NLRP3 inflammatory pathway were signifi-cantly decreased(P＜0.01),the numbers of GFAP and Iba-1 positive cells were significantly decreased(P＜0.01),the number of synapses of PC increased,the length of PSD increased,the thickness increased,and the morphology of cell membranes,cell nuclei,and mitochondria was improved(P＜0.05).Conclusion:DIP alleviates ethanol-induced mice cerebellar PC damage and mice motor dysfunction,improves ultrastructural abnormalities,and suppresses PC pyroptosis by inhibiting the NLRP3/GSDMD/IL-1β inflammatory pathway.