细胞焦亡在T-2毒素致大鼠关节软骨损伤中的作用研究  

The role of pyroptosis in rat articular cartilage injury induced by T-2 toxin

作　　者：董鹤萱 张鑫 李昊楠 齐芳 于倩 姜红 林步逸 于钧 Dong Hexuan;Zhang Xin;Li Haonan;Qi Fang;Yu Qian;Jiang Hong;Lin Buyi;Yu Jun(Institute of Kashin-Beck Disease,Center for Endemic Disease Control,Chinese Center for Disease Control and Prevention,Harbin Medical University,Harbin 150081,China)

机构地区：[1]哈尔滨医科大学中国疾病预防控制中心地方病控制中心大骨节病防治研究所,哈尔滨150081

出　　处：《中华地方病学杂志》2025年第1期10-16,共7页Chinese Journal of Endemiology

基　　金：国家自然科学基金(81972983,82473744)。

摘　　要：目的探讨细胞焦亡在T-2毒素致大鼠关节软骨损伤中的作用。方法选择145只SPF级Wistar雄性大鼠,按体重(50~70 g)随机分为空白对照组(n=45)、溶剂对照组(n=45)、T-2毒素组(n=55),T-2毒素组和溶剂对照组分别给予100 ng·g^(-1)·d^(-1)T-2毒素、等量无水乙醇溶剂灌胃处理,空白对照组常规饲养;分别于干预6、12、24周时,每组各处死15只大鼠,收集大鼠双侧膝关节。采用苏木素-伊红染色观察大鼠膝关节软骨病理改变;末端脱氧核苷酸转移酶介导的dUTP原位切口末端标记(TdT-mediated dUTP nick-end labeling,TUNEL)染色检测软骨细胞损伤情况;蛋白质印迹法检测焦亡执行蛋白消皮素D(gasdermin D,GSDMD)及其活化的N端结构域(cleaved N-terminal of gasdermin D,GSDMD-N)、NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、半胱氨酸天冬氨酸蛋白酶1(cysteinyl aspartate specific proteinase 1,Caspase-1)、白细胞介素1β(interleukin 1β,IL-1β)、白细胞介素18(interleukin 18,IL-18)、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing CARD,ASC)表达情况。结果干预6、12、24周时,T-2毒素组大鼠膝关节软骨组织出现软骨细胞数量减少、死亡等不同程度损伤。干预24周时,空白对照组、溶剂对照组、T-2毒素组软骨细胞TUNEL染色阳性率分别为(1.28±0.45)%、(0.73±0.27)%、(4.01±2.37)%,组间比较差异有统计学意义(F=6.11,P=0.036);且T-2毒素组高于空白对照组(P<0.05)。干预24周时,空白对照组、溶剂对照组、T-2毒素组NLRP3、Caspase-1、GSDMD、GSDMD-N、IL-1β蛋白表达水平比较,差异均有统计学意义(F=3.81、11.81、6.74、3.71、155.49,P=0.044、0.003、0.023、0.036、0.001);且T-2毒素组均高于空白对照组(均P<0.05)。不同干预周期时,各组ASC、IL-18蛋白表达水平比较,差异均无统计学意义(F=0.78、0.93、3.73,2.26、0.88、0.11,均P>0.05)。结论NLRP3/Caspase-1/GSDObjective To investigate the role of pyroptosis in T-2 toxin induced articular cartilage injury.Methods A total of 145 SPF grade male Wistar rats were randomly divided into blank control group(n=45),solvent control group(n=45),and T-2 toxin group(n=55)based on body weight(50-70 g).The T-2 toxin group and the solvent control group were given 100 ng·g^(-1)·d^(-1)T-2 toxin and an equal amount of anhydrous ethanol by gavage,respectively;the blank control group was fed routinely.Fifteen rats from each group were euthanized at 6,12,and 24 weeks of intervention,and bilateral knee joints of the rats were collected.Pathological changes in rat knee articular cartilage were observed using hematoxylin and eosin staining.TdT-mediated dUTP nick-end labeling(TUNEL)staining was used to detect chondrocyte injury.Western blot was used to detect the protein expression of gasdermin D(GSDMD),cleaved N-terminal of gasdermin D(GSDMD-N),NOD like receptor thermal protein domain associated protein 3(NLRP3),cysteinyl aspartate specific proteinase 1(Caspase-1),interleukin 1β(IL-1β),interleukin 18(IL-18),and apoptosis-associated spike-like protein containing CARD(ASC).Results At 6,12,and 24 weeks of intervention,the T-2 toxin group rats showed varying degrees of damage to the knee articular cartilage tissue,including a decrease in the number of chondrocytes and death.At 24 weeks of intervention,the TUNEL staining positivity rates of chondrocytes in the blank control group,solvent control group,and T-2 toxin group were(1.28±0.45)%,(0.73±0.27)%,and(4.01±2.37)%,respectively,with statistically significant differences between the groups(F=6.11,P=0.036);and the T-2 toxin group was higher than the blank control group(P<0.05).At 24 weeks of intervention,there were statistically significant differences in the expression levels of NLRP3,Caspase-1,GSDMD,GSDMD-N,and IL-1βproteins among the blank control group,solvent control group,and T-2 toxin group(F=3.81,11.81,6.74,3.71,155.49,P=0.044,0.003,0.023,0.036,0.001);and the T-2 toxin group was highe

关 键 词：关节软骨 T-2毒素 细胞焦亡 

分 类 号：R684.1[医药卫生—骨科学]