中华实用儿科临床杂志
中華實用兒科臨床雜誌
중화실용인과림상잡지
Journal of Applied Clinical Pediatrics
2015年
1期
59-63
,共5页
吴祖波%彭华%白燕%林雯%张志泉%刘亚黎
吳祖波%彭華%白燕%林雯%張誌泉%劉亞黎
오조파%팽화%백연%림문%장지천%류아려
硫化氢%柯萨奇病毒B3%病毒性心肌炎%肿瘤坏死因子-α%白细胞介素-6%白细胞介素-1β%核转录因子-κB
硫化氫%柯薩奇病毒B3%病毒性心肌炎%腫瘤壞死因子-α%白細胞介素-6%白細胞介素-1β%覈轉錄因子-κB
류화경%가살기병독B3%병독성심기염%종류배사인자-α%백세포개소-6%백세포개소-1β%핵전록인자-κB
Hydrogen sulfide%Coxsackie virus B3%Viral myocarditis%Tumor necrosis factor-α%Interleukin-1β%Interleukin-6%Nuclear factor kappa-B
目的 观察硫化氢(H2S)新型缓释剂GYY4137对柯萨奇B3病毒感染的大鼠乳鼠心肌细胞炎性因子的影响,探讨其可能的作用机制.方法 培养乳鼠原代心肌细胞,建立病毒性心肌炎(VMC)体外模型,采用简单随机法随机分为正常组、模型组、模型+ 10 μmol/L GYY4137组、模型+50 μmol/L GYY4137组、模型+100 μmol/L GYY4137组.采用细胞增殖毒性实验(CCK-8)法检测各组心肌细胞活力.24h后,酶联免疫吸附法(ELISA)检测各组白细胞介素-6(IL-6)、IL-1β及肿瘤坏死因子-α(TNF-α)的表达.Western blot法检测各组核转录因子-κB(NF-κB)抑制蛋白(Iκβoα)、细胞核因子蛋白(NF-κB p65)的表达.结果 不同浓度的GYY4137对各组心肌细胞活力无明显影响,各组心肌细胞活力均在80％以上;与正常组比较,CVB3感染各组IL-6、IL-1β、TNF-α[(175.80 ±5.05) ng/L,(25.80±1.97) ng/L,(65.33 ±3.51) ng/L]及胞核NF-κB p65表达显著升高(P＜0.01),Iκβα及胞质NF-κB p65表达降低(P＜0.01);与CVB3组比较,模型+10 μmol/L GYY4137组、模型+ 50 μmol/L GYY4137组、模型+100 μmol/L GYY4137组IL-6、IL-1β、TNF-α及胞核NF-κB p65表达明显降低,而Iκβα及胞质NF-κB p65表达增加(P＜0.01).结论 GYY4137可能通过抑制NF-κB信号转导通路,进而抑制柯萨奇B3病毒感染大鼠心肌细胞的炎性细胞因子的释放.
目的 觀察硫化氫(H2S)新型緩釋劑GYY4137對柯薩奇B3病毒感染的大鼠乳鼠心肌細胞炎性因子的影響,探討其可能的作用機製.方法 培養乳鼠原代心肌細胞,建立病毒性心肌炎(VMC)體外模型,採用簡單隨機法隨機分為正常組、模型組、模型+ 10 μmol/L GYY4137組、模型+50 μmol/L GYY4137組、模型+100 μmol/L GYY4137組.採用細胞增殖毒性實驗(CCK-8)法檢測各組心肌細胞活力.24h後,酶聯免疫吸附法(ELISA)檢測各組白細胞介素-6(IL-6)、IL-1β及腫瘤壞死因子-α(TNF-α)的錶達.Western blot法檢測各組覈轉錄因子-κB(NF-κB)抑製蛋白(Iκβoα)、細胞覈因子蛋白(NF-κB p65)的錶達.結果 不同濃度的GYY4137對各組心肌細胞活力無明顯影響,各組心肌細胞活力均在80％以上;與正常組比較,CVB3感染各組IL-6、IL-1β、TNF-α[(175.80 ±5.05) ng/L,(25.80±1.97) ng/L,(65.33 ±3.51) ng/L]及胞覈NF-κB p65錶達顯著升高(P＜0.01),Iκβα及胞質NF-κB p65錶達降低(P＜0.01);與CVB3組比較,模型+10 μmol/L GYY4137組、模型+ 50 μmol/L GYY4137組、模型+100 μmol/L GYY4137組IL-6、IL-1β、TNF-α及胞覈NF-κB p65錶達明顯降低,而Iκβα及胞質NF-κB p65錶達增加(P＜0.01).結論 GYY4137可能通過抑製NF-κB信號轉導通路,進而抑製柯薩奇B3病毒感染大鼠心肌細胞的炎性細胞因子的釋放.
목적 관찰류화경(H2S)신형완석제GYY4137대가살기B3병독감염적대서유서심기세포염성인자적영향,탐토기가능적작용궤제.방법 배양유서원대심기세포,건립병독성심기염(VMC)체외모형,채용간단수궤법수궤분위정상조、모형조、모형+ 10 μmol/L GYY4137조、모형+50 μmol/L GYY4137조、모형+100 μmol/L GYY4137조.채용세포증식독성실험(CCK-8)법검측각조심기세포활력.24h후,매련면역흡부법(ELISA)검측각조백세포개소-6(IL-6)、IL-1β급종류배사인자-α(TNF-α)적표체.Western blot법검측각조핵전록인자-κB(NF-κB)억제단백(Iκβoα)、세포핵인자단백(NF-κB p65)적표체.결과 불동농도적GYY4137대각조심기세포활력무명현영향,각조심기세포활력균재80％이상;여정상조비교,CVB3감염각조IL-6、IL-1β、TNF-α[(175.80 ±5.05) ng/L,(25.80±1.97) ng/L,(65.33 ±3.51) ng/L]급포핵NF-κB p65표체현저승고(P＜0.01),Iκβα급포질NF-κB p65표체강저(P＜0.01);여CVB3조비교,모형+10 μmol/L GYY4137조、모형+ 50 μmol/L GYY4137조、모형+100 μmol/L GYY4137조IL-6、IL-1β、TNF-α급포핵NF-κB p65표체명현강저,이Iκβα급포질NF-κB p65표체증가(P＜0.01).결론 GYY4137가능통과억제NF-κB신호전도통로,진이억제가살기B3병독감염대서심기세포적염성세포인자적석방.
Objective To study the protective effect of GYY4137 on rat myocardial cells infected by Coxsackie virus B3 (CVB3) and its signal transduction mechanism.Methods Cardiomyocytes were treated with different concentrations of GYY4137(10,50,100 μmol/L) for 24 hours before addition of 100 TCID50 CVB3 for 2 hours before serum-free conditions.After treatment,the cell viability was ascertained with Cell Counting Kit-8 (CCK-8) assay.At the same time,the levels of tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β),interleukin-6 (IL-6) in the supernatants were analyzed by enzyme-linked immunosorbent assay (ELISA).Western blot was used to study the expression of nuclear factor kappa-B (NF-κB) protein and the inhibitory subunit of (IκBα) in myocardial cells.Results After exposure of cardiomyocytes to GYY4137 with different concentration (10,50,100 μmol/L)for 24 hours cell viability had no change.The NF-κB expression and the levels of TNF-α,IL-1β,IL-6 [(175.80 ± 5.05) ng/L,(25.80 ± 1.97) ng/L,(65.33 ± 3.51) ng/L] in the GYY4137-treated CVB3 infection group were significantly reduced when compared with untreated CVB3 infection group (P ＜ 0.01),respectively.Compared with the normal group,the GYY4137 concentration-dependently restrained the CVB3-mediated IκBα degradation(P ＜ 0.01).Conclusions GYY4137 exerts anti-inflammatory effects in CVB3-infected cardiomyocytes.This anti-inflammatory mechanism may be associated with suppression of the activation of the NF-κB.
