中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
12期
2786-2789
,共4页
薛华明%涂意辉%马童%文涛%夏志道%梅炯
薛華明%塗意輝%馬童%文濤%夏誌道%梅炯
설화명%도의휘%마동%문도%하지도%매형
软骨细胞%增殖%地塞米松%乳铁蛋白
軟骨細胞%增殖%地塞米鬆%乳鐵蛋白
연골세포%증식%지새미송%유철단백
Chondrocytes%Proliferation%Dexamethasone%Lactoferrin
目的 观察地塞米松(Dex)对人骨关节炎软骨细胞(HACs)增殖力和活力的影响以及乳铁蛋白(LF)的干预作用.方法 体外培养HACs,在浓度为25 mg/L的Dex和不同浓度LF(12.5、25.0、50.0、100.0、200.0 mg/L)的培养基中培养72 h,采用噻唑蓝(MTT)法测定HACs的增殖力,LIVE/DEAD染色法检测HACs存活力,实时定量聚合酶链反应(Real-time PCR)和免疫细胞化学染色法检测磷酸化细胞外信号调节激酶(p-ERK)/ERK基因的表达.结果 Dex显著抑制HACs的增殖力及活力(P＜0.01),并伴随着ERK mRNA和p-ERK/ERK蛋白表达明显下调(P＜0.05).单纯Dex及各浓度LF组均能以浓度依赖方式显著抑制Dex诱发的HACs增殖力下降,各组分别为(38.9±1.3)％、(55.6±3.1)％、(94.4±1.4)％、(111.1±3.2)％、(127.8±2.5)％和(150.0±4.3)％(P＜0.01).12.5 mg/L的LF并未明显促进软骨细胞的存活力(P＞0.05),而25.0～ 200.0 mg/L的LF可以抑制Dex诱发的HACs的存活力下降,各组分别为(73.2±3.7)％、(82.1±5.1)％、(89.1±4.5)％和(95.5±2.9)％(P＜0.01),呈明显的浓度依赖性,同时ERK基因mRNA和p-ERK/ERK蛋白表达的下调得到显著改善(P＜0.05).结论 LF可阻止Dex诱导的HACs增殖力及活力抑制,其作用机制可能与明显上调增殖基因ERK的mRNA和蛋白水平的表达有关.
目的 觀察地塞米鬆(Dex)對人骨關節炎軟骨細胞(HACs)增殖力和活力的影響以及乳鐵蛋白(LF)的榦預作用.方法 體外培養HACs,在濃度為25 mg/L的Dex和不同濃度LF(12.5、25.0、50.0、100.0、200.0 mg/L)的培養基中培養72 h,採用噻唑藍(MTT)法測定HACs的增殖力,LIVE/DEAD染色法檢測HACs存活力,實時定量聚閤酶鏈反應(Real-time PCR)和免疫細胞化學染色法檢測燐痠化細胞外信號調節激酶(p-ERK)/ERK基因的錶達.結果 Dex顯著抑製HACs的增殖力及活力(P＜0.01),併伴隨著ERK mRNA和p-ERK/ERK蛋白錶達明顯下調(P＜0.05).單純Dex及各濃度LF組均能以濃度依賴方式顯著抑製Dex誘髮的HACs增殖力下降,各組分彆為(38.9±1.3)％、(55.6±3.1)％、(94.4±1.4)％、(111.1±3.2)％、(127.8±2.5)％和(150.0±4.3)％(P＜0.01).12.5 mg/L的LF併未明顯促進軟骨細胞的存活力(P＞0.05),而25.0～ 200.0 mg/L的LF可以抑製Dex誘髮的HACs的存活力下降,各組分彆為(73.2±3.7)％、(82.1±5.1)％、(89.1±4.5)％和(95.5±2.9)％(P＜0.01),呈明顯的濃度依賴性,同時ERK基因mRNA和p-ERK/ERK蛋白錶達的下調得到顯著改善(P＜0.05).結論 LF可阻止Dex誘導的HACs增殖力及活力抑製,其作用機製可能與明顯上調增殖基因ERK的mRNA和蛋白水平的錶達有關.
목적 관찰지새미송(Dex)대인골관절염연골세포(HACs)증식력화활력적영향이급유철단백(LF)적간예작용.방법 체외배양HACs,재농도위25 mg/L적Dex화불동농도LF(12.5、25.0、50.0、100.0、200.0 mg/L)적배양기중배양72 h,채용새서람(MTT)법측정HACs적증식력,LIVE/DEAD염색법검측HACs존활력,실시정량취합매련반응(Real-time PCR)화면역세포화학염색법검측린산화세포외신호조절격매(p-ERK)/ERK기인적표체.결과 Dex현저억제HACs적증식력급활력(P＜0.01),병반수착ERK mRNA화p-ERK/ERK단백표체명현하조(P＜0.05).단순Dex급각농도LF조균능이농도의뢰방식현저억제Dex유발적HACs증식력하강,각조분별위(38.9±1.3)％、(55.6±3.1)％、(94.4±1.4)％、(111.1±3.2)％、(127.8±2.5)％화(150.0±4.3)％(P＜0.01).12.5 mg/L적LF병미명현촉진연골세포적존활력(P＞0.05),이25.0～ 200.0 mg/L적LF가이억제Dex유발적HACs적존활력하강,각조분별위(73.2±3.7)％、(82.1±5.1)％、(89.1±4.5)％화(95.5±2.9)％(P＜0.01),정명현적농도의뢰성,동시ERK기인mRNA화p-ERK/ERK단백표체적하조득도현저개선(P＜0.05).결론 LF가조지Dex유도적HACs증식력급활력억제,기작용궤제가능여명현상조증식기인ERK적mRNA화단백수평적표체유관.
Objective To examine the effect of dexamethasone (Dex) on proliferation and viability of chondrocytes and preventive effect of lactoferrin (LF).Methods Human articular chondrocytes (HACs) were incubated in cell culture media containing 25 mg/L Dex and different concentrations of LF (12.5,25.0,50.0,100.0,and 200.0 mg/L) for 72 h.Proliferation was assessed by methyl thiazol tetrazolium (MTT).Viability was determined by Live/Dead assay.The mRNA and protein expression of extracellular signal-regulated kinase (ERK) was detected by real-time quantitative polymerase chain reaction (Real-time PCR) and immunocytochemistry,respectively.Results Dex significantly inhibited the proliferation (P ＜ 0.01) and viability (P ＜ 0.01) of HACs and the expression levels of ERK mRNA and protein (P ＜0.05) were significantly down-regulated.LF could inhibite Dex-induced proliferation reduction of HACs from 0 mg/L to 200.0 mg/L,with the proliferation rate at LF concentrations of 12.5,25.0,50.0,100.0,and 200.0 mg/L being (38.9±1.3)％,(55.6±3.1)％,(94.4±1.4)％,(111.1±3.2)％,(127.8 ± 2.5) ％ and (150.0 ± 4.3) ％ respectively (P ＜ 0.01).In addition,Dex-induced viability reduction of HACs was significantly inhibited by LF from 25.0 mg/L to 200.0 mg/L in a dose-dependent manner with (73.2 ± 3.7) ％,(82.1 ± 5.1) ％,(89.1 ± 4.5) ％ and (95.5 ± 2.9) ％ respectively (P ＜ 0.01).Treatment of HACs with LF also resulted in a significant increase in the expression of ERK mRNA and p-ERK/ERK protein in cultured HACs (P ＜ 0.05).Conclusion LF can afford preventive effects against the inhibition of proliferation and viability of HACs induced by Dex,which may be related to the significant up-regulation of the p-ERK/ERK gene expression.
