基于NF-κB/NLRP3/caspase-1信号轴探讨牛膝皂苷Ⅰ抑制软骨细胞焦亡的研究  

Research of Achyranthoside Ⅰ inhibiting pyroptosis in chondrocytes based on the NF-kB/NLRP3/caspase-1 signaling axis

作　　者：刘泽萱 韩易言 关雪峰 张宇 戴俭宇[1,2] LIU Ze-xuan;HAN Yi-yan;GUAN Xue-feng;ZHANG Yu;DAI Jian-yu(College of Acupuncture and Massage,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning Province,China;Key Laboratory of Acupuncture and Meridian Biology of Education Department of Liaoning,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning Province,China;Party and Government Office,Shenyang Pharmaceutical University,Shenyang 110016,Liaoning Province,China)

机构地区：[1]辽宁中医药大学针灸推拿学院,辽宁沈阳110847 [2]辽宁中医药大学辽宁省教育厅针灸生物学重点实验室,辽宁沈阳110847 [3]沈阳药科大学党政办公室,辽宁沈阳110016

出　　处：《中国临床药理学杂志》2025年第2期198-202,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家重点研发计划基金资助项目(2021YFC1712800);国家资助博士后研究人员计划基金资助项目(GZC20231025);辽宁省科学技术计划项目创新能力提升联合基金资助项目(2022-NLTS-13-02);辽宁省教育厅基本科研基金资助项目(JYTQN2023458)。

摘　　要：目的探索牛膝皂苷Ⅰ通过核因子κB(NF-κB)/DON样受体蛋白3(NLRP3)/半胱氨酸蛋白酶-1(caspase-1)信号传导途径抑制软骨细胞发生焦亡的作用机制。方法将小鼠原代软骨细胞分为空白组(加入等体积的磷酸盐缓冲溶液)、模型组[10 ng·mL^(-1)白细胞介素1β(IL-1β)]、对照组(10ng·mL^(-1)IL-1β+20μmol·L-1塞来昔布)和实验组(10ng·mL^(-1)IL-1β+3μg·mL^(-1)牛膝皂苷Ⅰ)。干预24h后,用细胞计数试剂盒8法检测细胞增殖能力,用酶联免疫吸附试验法检测细胞上清液中超氧化物歧化酶(SOD)、丙二醛(MDA)、IL-1和IL-6水平,用蛋白质印迹法检测NF-κBp65、NLRP3和caspase-1的表达水平。结果实验组、对照组、模型组和空白组的细胞凋亡率分别为(13.34±0.61)%、(15.64±1.01)%、(21.81±1.10)%和0,SOD水平分别为(147.03±16.49)、(130.09±7.33)、(122.03±10.71)和(164.40±22.74)nU·mL^(-1),MDA水平分别为(6.43±0.71)、(7.63±1.01)、(8.89±1.84)和(5.69±0.81)nmol·L-1,IL-1水平分别为(338.69±40.95)、(361.78±32.15)、(391.44±30.59)和(289.23±25.19)pg·mL^(-1),IL-6水平分别为(89.96±8.81)、(101.10±11.59)、(120.39±14.71)和(60.29±6.03)pg·mL^(-1),NF-κBp65蛋白相对表达水平分别为0.68±0.05、0.97±0.05、1.26±0.05和0.57±0.05,NLRP3蛋白相对表达水平分别为0.71±0.08、1.02±0.10、1.50±0.06和0.31±0.05,caspase-1蛋白相对表达水平分别为0.70±0.07、1.29±0.08、1.66±0.07和0.51±0.07。实验组的上述指标与模型组相比,在统计学上差异均有统计学意义(均P<0.05)。结论牛膝皂苷Ⅰ可以改善IL-1β诱导软骨细胞炎症的氧化应激状态,降低NF-κB信号通路相关蛋白的表达,减少caspase-1依赖性细胞焦亡的发生,从而对软骨细胞起到保护作用。Objective To investigate the mechanism of Achyranthoside Ⅰ inhibits pyroptosis in chondrocytes through the nuclear factor-kB(NF-kB)/NOD receptor protein structure domain related proteins 3(NLRP3)/cystine containing aspartate specific proteins-1(caspase-1)signaling pathway.Methods Primary mouse chondrocytes were divided into blank group(phosphate buffered solution with the same volume),model group[10 ng·mL^(-1) interleukin-1β(IL-1β)],control group(10 ng·mL^(-1) IL-1β+20μmol·L^(-1)celecoxib)and experimental group(10 ng·mL^(-1)IL-1β+3μg·mL^(-1)Achyranthoside I).After 24 hours of intervention,the cell proliferation was measured by cell counting kit 8,the levels of superoxide dismutase(SOD),malondialdehyde(MDA),IL-1 and IL-6 were detected by enzyme-linked immunosorbent assay,the protein expression levels of NF-kB p65,NLRP3 and caspase-1 were detected by Western Blot.ResultsTThe apoptosis rates in experimental,control,model and blank groups were(13.34±0.61)%,(15.64±1.01)%,(21.81±1.10)%and 0;the S0D levels were(147.03±16.49),(130.09±7.33),(122.03±10.71)and(164.40±22.74)nU·mL^(-1)1;the MDA levels were(6.43±0.71),(7.63±1.01),(8.89±1.84)and(5.69±0.81)nmol·L^(-1)1;the IL-1 levels were(338.69±40.95),(361.78±32.15),(391.44±30.59)and(289.23±25.19)pg·mL^(-1);the IL-6 levels were(89.96±8.81),(101.10±11.59),(120.39±14.71)and(60.29±6.03)pg·mL^(-1);the relative expression levels of NF-kB p65 were 0.68±0.05,0.97±0.05,1.26±0.05 and 0.57±0.05;the relative expression levels of NLRP3 were 0.71±0.08,1.02±0.10,1.50±0.06 and 0.31±0.05;the relative expression levels of caspase-1 were 0.70±0.07,1.29±0.08,1.66±0.07 and 0.51±0.07,respectively.Compared with the model group,the differences of above indexes were statistically significant in the experimental group(all P<0.05).Conclusion Achyranthoside Ⅰ can improve the oxidative stress status induced by IL-1βin chondrocytes,reduce the expression of proteins related to the NF-kB signaling pathway,and thereby decrease the occurrence of caspase-1 dep

关 键 词：牛膝皂苷Ⅰ 软骨细胞 氧化应激 半胱氨酸蛋白酶-1依赖性 细胞焦亡 

分 类 号：R28[医药卫生—中药学]