摘要
目的探讨CXC趋化因子配体14(CXCL14)对高糖暴露环境中脂肪细胞焦亡的影响。方法利用"鸡尾酒法"诱导3T3-L1细胞分化为成熟脂肪细胞,用5.5 mmol/L低糖(NG)或25 mmol/L高糖(HG)葡萄糖培养基培养脂肪细胞24 h;HG环境下用不同浓度CXCL14处理3T3-L1细胞不同时间。Western blot检测消化道皮肤素(GSDMD)、核苷酸结合寡聚化结构样受体蛋白3(NLRP3)、天冬氨酸蛋白水解酶-1(Caspase-1)蛋白、白细胞介素-6(IL-6)蛋白表达水平。实时荧光定量PCR检测GSDMD、NLRP3、白细胞介素-1β(IL-1β)mRNA转录水平。LDH测定试剂盒检测脂肪细胞上清液中乳酸脱氢酶(LDH)活力;Annexin V-FITC荧光检测细胞死亡情况;CCK-8法检测各组细胞增殖活力。结果高糖环境下脂肪细胞焦亡发生率升高,CXCL14处理可提高脂肪细胞增殖活力,但脂肪细胞焦亡相关指标却受到CXCL14浓度梯度的不同影响。50 nmol/L CXCL14处理可降低高糖环境下脂肪细胞GSDMD、NLRP3、IL-1βmRNA以及NLRP3蛋白的表达,下调脂肪细胞LDH活力,减少Annexin V-FITC荧光染色细胞死亡率,但25、100、200 nmol/L CXCL14对其焦亡的指标却呈反向趋势。并且50 nmol/L CXCL14干预后脂肪细胞NLRP3、Caspase-1蛋白随干预时间的延长呈先下降再上升趋势。结论CXCL14对高糖环境中脂肪细胞焦亡的影响与其浓度存在相关性。
Aim To investigate the effect of C-X-C motif chemokine ligand 14(CXCL14)on pyroptosis of adi-pocytes exposed to high glucose.Methods 3 T3-L1 cell lines were induced to differentiate into mature adipocytes byclassical‘cocktail’method.Adipocytes were cultured in 5.5 mmol/L(NG)or 25 mmol/L(HG)glucose medium for 24 hours.3 T3-L1 cells were treated with different concentrations of CXCL14 in HG environment for different time,and theprotein expression levels of gasdermin D(GSDMD),nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),Caspase-1 and interleukin-6(IL-6)were detected by Western blot.The mRNA transcription levels of GSD-MD,NLRP3 and interleukin-1β(IL-1β)were detected by real-time fluorescence quantitative PCR.Lacticdehydrogenase(LDH)activity in supermatant of adipocytes were detected by LDH assay kit.Cell death was detected byAnnexin V-FITC fluorescence.And the proliferation activity of each group was detected by CCK-8 method.Results The incidence of adipocyte pyroptosis in high glucose environment increased.CXCL14 treatment can improve the prolifer-ation activity of adipocytes.However,adipocyte pyroptosis-related indicators are differently affected by the concentrationgradient of CXCL14.50 nmol/L CXCL14 treatment can reduce the expression of GSDMD,NLRP3,IL-1βmRNA andNLRP3 protein in adipocytes under high glucose environment,down-regulate the LDH activity of adipocytes and reduce An-nexin V-FITC fluorescent staining cell death rate.But 25,100,200 nmol/L CXCL14 treatment groups show a reverse trend.And the NLRP3 and Caspase-1 proteins of adipocytes treated with 50 nmol/L CXCL14 decreased first and then in-creased with the prolongation of intervention time.Conclusion The effect of CXCL14 on the pyroptosis of adipocytesin a high glucose environment is related to its concentration.
作者
侯丽娜
刘嘉
李亚兰
孙振
张莉莉
王中群
HOU Lina;LIU Jia;LI Yalan;SUN Zhen;ZHANG Lili;WANG Zhongqun(Department of Cardiology,Affiliated Hospital of Jiangsu University,Zhenjiang,Jiangsu 212001,China)
出处
《中南医学科学杂志》
CAS
2022年第1期7-12,共6页
Medical Science Journal of Central South China
基金
国家自然科学基金资助项目(82070455,81770450)
江苏省自然科学基金资助项目(BK20201225)。
