CA72-4对单核细胞IL-1β和痛风急性发作的调控作用OA
Regulatory Effect of CA72-4 on Monocyte IL-1β and Acute Gout Attack
目的:探究糖类抗原(carbohydrate antigen 72-4,CA72-4)对单核细胞白细胞介素-1β(interleukin-1β,IL-1β)和痛风急性发作的调控作用.方法:收集淄博市中心医院痛风专病门诊收治的急性痛风患者外周血浆,分析CA72-4 含量,获取CA72-4≥50 U/mL的患者血浆,随机分为no-CA72-4 组(n=37)和CA72-4 组(n=38),no-CA72-4 组采用DynaMag磁架去除血浆中的CA72-4,CA72-4 组不做处理,将处理前后的血浆分别诱导THP1 细胞,收集诱导后THP1细胞培养液和细胞,采用酶联免疫法(enzyme-linked immunosorbent assay,ELISA)检测THP1 细胞和培养液中IL-1β表达水平;将小鼠背部皮下注射灭菌过滤空气,造成皮下气囊,随后将尿酸盐(monosodium urate,MSU)结晶注入气囊,构建痛风性滑膜炎模型,将模型小鼠分成试验组(n=10)和对照组(n=10),试验组腹腔注射 1.5×1010 PFU/mL腺病毒(adenovirus,Ad)-CA72-4,对照组注射照腺病毒Ad-ctr,收集小鼠血浆和滑膜液,采用CA72-4 检测试剂盒检测小鼠血浆和滑膜液中CA72-4 含量;采用ELISA法检测小鼠血浆中IL-1β和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达水平,实时定量荧光聚合酶链反应和蛋白质印迹法(Western blot)检测各组小鼠血浆IL-1β、核苷酸结合寡聚化结构域样受体蛋白 3(nucleotide binding oligomerization domain-like receptor protein 3,NLRP3)炎症小体基因和蛋白表达水平.结果:与CA72-4组相比,no-CA72-4 组THP1 细胞和培养液中IL-1β明显升高,差异有统计学意义(P<0.001);与对照组相比,试验组小鼠血浆和滑膜液中CA72-4 表达水平显著高,差异有统计学意义(P<0.001);与对照组相比,试验组小鼠血浆中IL-1β和TNF-α表达均显著高,差异有统计学意义(P<0.001);与对照组相比,试验组小鼠血浆中IL-1β、NALP3 基因和蛋白表达均显著升高,差异有统计学意义(P<0.001).结论:CA72-4 能增强单核细胞炎症因子分泌,加重痛风炎症反应,其具体的作用机制需进一步研究.
Objective:To investigate the regulatory effect of carbohydrate antigen 72-4(CA72-4)on monocyte interleukin 1β(IL-1β)and acute gout attack.Method:The peripheral plasma of acute gout patients who admitted to the Gout Specialist Clinic of Zibo Central Hospital was collected,and the content of CA72-4 was analyzed to obtain the plasma of patients with CA72-4≥50 U/mL,which was randomly divided into no-CA72-4 group(n=37)and CA72-4 group(n=38),no-CA72-4 group used DynaMag magnetic frame to remove CA72-4 from the plasma,while CA72-4 group was not treated,THP1 cells were induced by plasma before and after treatment,the induced THP1 cell culture medium and cells were collected,and the expression level of IL-1β in THP1 cells and culture medium was detected by enzyme-linked immunoassay(ELISA).Subcutaneous injection of sterilized and filtered air into the back of mice resulted in subcutaneous air sac,and then monosodium urate(MSU)crystals were injected into the air sac to construct gouty synovitis model,the model mice were divided into the experimental group(n=10)and the control group(n=10),the experimental group was intraperitoneally injected with 1.5×1010 PFU/mL overexpressed adenovirus(Ad)-CA72-4,while the control group was injected with adenovirus Ad-ctr,plasma and synovial fluid of mice were collected,and CA72-4 detection kit was used to detect CA72-in the plasma and synovial fluid of mice.The expression levels of IL-1β and tumor necrosis factor-α(TNF-α)in plasma of mice were detected by ELISA,the gene and protein expression levels of IL-1β and nucleotide binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome in plasma of mice in each group were detected by real-time quantitative fluorescence polymerase chain reaction and Western blot.Result:Compared with CA72-4 group,IL-1β in THP1 cells and culture medium in no-CA72-4 group were significantly increased,and the differences were statistically significant(P<0.001);compared with the control group,the expression levels of CA72-4 in plasma and synovial fluid of mice in the experimental group were significantly increased,and the differences were statistically significant(P<0.001);compared with the control group,the expressions of IL-1β and TNF-α in the plasma of the experimental group were significantly increased,and the differences were statistically significant(P<0.001);compared with the control group,the gene and protein expressions of IL-1β and NALP3 in the plasma of mice in the experimental group were significantly increased,and the differences were statistically significant(P<0.001).Conclusion:CA72-4 can enhance the secretion of inflammatory factors in monocytes and aggravate the inflammatory response of gout,and its specific mechanism needs further study.
胡红蕾;郑宁宁;聂宵;白雪山
淄博市中心医院 山东 淄博 255000
CA72-4痛风单核细胞核苷酸结合寡聚化结构域样受体蛋白3炎症小体
Carbohydrate antigen 72-4GoutMonocyteNucleotide binding oligomerization domain-like receptor protein 3 inflammatory corpuscles
《中外医学研究》 2024 (004)
5-9 / 5
山东省医药卫生科技发展计划项目(202103060479)
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