中国综合临床
中國綜閤臨床
중국종합림상
CLINICAL MEDICINE OF CHINA
2012年
7期
704-707
,共4页
骨桥蛋白%柯萨奇B3病毒%心肌成纤维细胞%Ⅰ型胶原
骨橋蛋白%柯薩奇B3病毒%心肌成纖維細胞%Ⅰ型膠原
골교단백%가살기B3병독%심기성섬유세포%Ⅰ형효원
Osteopontin%Coxsackievirus B3%Myocardial fibroblasts%Collagen type Ⅰ
目的 检测柯萨奇病毒B3( CVB3)对大鼠心肌成纤维细胞骨桥蛋白(OPN)及Ⅰ型胶原表达的影响,探讨OPN在病毒性心肌炎的可能发病机制.方法 用重复感染率(MOI)为0.5 PFU/cell的CVB3感染体外培养的大鼠心肌成纤维细胞,细胞分对照组(12 h)、病毒组(感染后12 h、ld、2d及3 d).用RT-PCR及Western Blot、免疫组化检测OPN及Ⅰ型胶原表达,并将OPN表达与Ⅰ型胶原表达进行直线相关分析.结果 (1)RT-PCR测得对照组12h、病毒组12 h、1d、2d及3d5组OPN/β-actin吸光度比值分别为:0.38±0.06、0.56±0.06、0.72 ±0.05、0.98 ±0.06、0.86 ±0.02；Western Blot测得上述5组的OPN/β-actin灰度值分别为:0.26±0.03、0.36±0.03、0.52±0.04、0.76±0.05、0.62±0.02;感染后12 h,OPN表达即增高,于2d达到高峰,3d呈下降趋势,OPN/β-actin吸光度比值、灰度值比较差异均有统计学意义(F值分别为74.965、53.004,P均＜0.05)；(2)RT-PCR测得心肌成纤维细胞对照组12 h、病毒组12h、1d、2d、3dⅠ型胶原/β-actin吸光度比值分别为:1.12±0.03、1.18±0.01、1.22±0.02、1.33±0.02、1.28±0.03,感染后12h Ⅰ型胶原表达即增高,于2d达到高峰,3d呈下降趋势,差异有统计学意义(F=38.241,P＜0.05)；(3)OPN表达与Ⅰ型胶原有正相关关系(r=0.948,P＜0.001).结论 CVB3可诱导心肌成纤维细胞Ⅰ型胶原及OPN表达,OPN表达与Ⅰ型胶原表达呈正相关,提示OPN可能促进心肌成纤维细胞Ⅰ型胶原表达,参与病毒性心肌炎心肌纤维化的发病机制.
目的 檢測柯薩奇病毒B3( CVB3)對大鼠心肌成纖維細胞骨橋蛋白(OPN)及Ⅰ型膠原錶達的影響,探討OPN在病毒性心肌炎的可能髮病機製.方法 用重複感染率(MOI)為0.5 PFU/cell的CVB3感染體外培養的大鼠心肌成纖維細胞,細胞分對照組(12 h)、病毒組(感染後12 h、ld、2d及3 d).用RT-PCR及Western Blot、免疫組化檢測OPN及Ⅰ型膠原錶達,併將OPN錶達與Ⅰ型膠原錶達進行直線相關分析.結果 (1)RT-PCR測得對照組12h、病毒組12 h、1d、2d及3d5組OPN/β-actin吸光度比值分彆為:0.38±0.06、0.56±0.06、0.72 ±0.05、0.98 ±0.06、0.86 ±0.02；Western Blot測得上述5組的OPN/β-actin灰度值分彆為:0.26±0.03、0.36±0.03、0.52±0.04、0.76±0.05、0.62±0.02;感染後12 h,OPN錶達即增高,于2d達到高峰,3d呈下降趨勢,OPN/β-actin吸光度比值、灰度值比較差異均有統計學意義(F值分彆為74.965、53.004,P均＜0.05)；(2)RT-PCR測得心肌成纖維細胞對照組12 h、病毒組12h、1d、2d、3dⅠ型膠原/β-actin吸光度比值分彆為:1.12±0.03、1.18±0.01、1.22±0.02、1.33±0.02、1.28±0.03,感染後12h Ⅰ型膠原錶達即增高,于2d達到高峰,3d呈下降趨勢,差異有統計學意義(F=38.241,P＜0.05)；(3)OPN錶達與Ⅰ型膠原有正相關關繫(r=0.948,P＜0.001).結論 CVB3可誘導心肌成纖維細胞Ⅰ型膠原及OPN錶達,OPN錶達與Ⅰ型膠原錶達呈正相關,提示OPN可能促進心肌成纖維細胞Ⅰ型膠原錶達,參與病毒性心肌炎心肌纖維化的髮病機製.
목적 검측가살기병독B3( CVB3)대대서심기성섬유세포골교단백(OPN)급Ⅰ형효원표체적영향,탐토OPN재병독성심기염적가능발병궤제.방법 용중복감염솔(MOI)위0.5 PFU/cell적CVB3감염체외배양적대서심기성섬유세포,세포분대조조(12 h)、병독조(감염후12 h、ld、2d급3 d).용RT-PCR급Western Blot、면역조화검측OPN급Ⅰ형효원표체,병장OPN표체여Ⅰ형효원표체진행직선상관분석.결과 (1)RT-PCR측득대조조12h、병독조12 h、1d、2d급3d5조OPN/β-actin흡광도비치분별위:0.38±0.06、0.56±0.06、0.72 ±0.05、0.98 ±0.06、0.86 ±0.02；Western Blot측득상술5조적OPN/β-actin회도치분별위:0.26±0.03、0.36±0.03、0.52±0.04、0.76±0.05、0.62±0.02;감염후12 h,OPN표체즉증고,우2d체도고봉,3d정하강추세,OPN/β-actin흡광도비치、회도치비교차이균유통계학의의(F치분별위74.965、53.004,P균＜0.05)；(2)RT-PCR측득심기성섬유세포대조조12 h、병독조12h、1d、2d、3dⅠ형효원/β-actin흡광도비치분별위:1.12±0.03、1.18±0.01、1.22±0.02、1.33±0.02、1.28±0.03,감염후12h Ⅰ형효원표체즉증고,우2d체도고봉,3d정하강추세,차이유통계학의의(F=38.241,P＜0.05)；(3)OPN표체여Ⅰ형효원유정상관관계(r=0.948,P＜0.001).결론 CVB3가유도심기성섬유세포Ⅰ형효원급OPN표체,OPN표체여Ⅰ형효원표체정정상관,제시OPN가능촉진심기성섬유세포Ⅰ형효원표체,삼여병독성심기염심기섬유화적발병궤제.
Objective To study the expression of osteopontin (OPN) and collagen type Ⅰ in the rat myocardial fibroblasts infected by coxsackievirus B3 (CVB3) and to explore the possible pathogenesis of OPN on viral myocarditis.Methods Cardiac fibroblasts were isolated from neonatal rat and cultured with an traditional method.The primary cultured rat myocardial fibroblasts were infected by CVB3 which multiplicity of infection (MOI) was 0.5 PFU/cell.The myocardial fibroblasts were divided into control group 12 h and CVB3 groups (infected after 12 h,1 d,2 d,3 d).The expression of OPN and collagen type Ⅰ were detected by RT-PCR,Western Blotting and immunohistochemistry.And the linear correlation between the expression of OPN and the expression of collagen type Ⅰ was analyzed.Results ( 1 ) The gray scale values of the OPN/β-actin in control group(12 h)and viral groups (12 h,1 d,2 d,3 d) were 0.38 ± 0.06,0.56 ± 0.06,0.72 ± 0.05,0.98 ± 0.06,0.86 ± 0.02 respectively with RT-PCR,and were 0.26 ± 0.03,0.36 ± 0.03,0.52 ± 0.04,0.76 ± 0.05,0.62 ± 0.02 respectively with Western Blotting.The expression of OPN was found to be increased after 12 h infection,reached to the maximum after 2 d infection and displayed a decreased tendency after 3 d infection.There was significant difference on the gray scale values of the OPN/β3-actin ( F=74.965,53.004,respectively,P ＜ 0.05 ).(2) The gray scale values of the collagen type Ⅰ/β-actin in control group (12 h)and viral groups (12 h,1 d,2d,3 d) were 1.12 ± 0.03,1.18 ± 0.01,1.22 ± 0.02,1.33 ± 0.02,1.28 ± 0.03,respectively with RTPCR.These results suggested that the collagen type Ⅰ expression started to increase at 12 h when infected by CVB3,reached to the maximum at 2 d,and then decreased after 3 d infection,the difference was significant ( F =38.241,P ＜ 0.05).(3) The expression of OPN was positively correlated with the expression of collagen type Ⅰ ( r=0.948,P ＜ 0.001 ).Conclusion CVB3 can induce the expression of OPN and collagen type Ⅰ in the rat myocardial fibroblasts and the expression of OPN and collagen type Ⅰ displays positive correlation.It suggests that OPN can promote the expression of collagen type Ⅰ in myocardial fibroblasts,and may play an important role in the pathogenesis of viral myocarditis.
