中华儿科杂志
中華兒科雜誌
중화인과잡지
Chinese Journal of Pediatrics
2013年
1期
42-46
,共5页
费文君%闫波%袁丽萍%张琴%胡波%鹿玲
費文君%閆波%袁麗萍%張琴%鬍波%鹿玲
비문군%염파%원려평%장금%호파%록령
紫癜,过敏性%内皮细胞%免疫球蛋白A%细胞凋亡
紫癜,過敏性%內皮細胞%免疫毬蛋白A%細胞凋亡
자전,과민성%내피세포%면역구단백A%세포조망
Purpura,sch(o)nlein-Henoch%Endothelial cells%Immunoglobulin A%Apoptosis
目的 探讨过敏性紫癜(HSP)患儿血清IgA1对脐静脉内皮细胞(HUVEC)凋亡的影响.方法 培养人HUVEC,分为HSP组、正常对照组和空白对照组.亲和层析法分离HSP患儿及正常儿童血清IgA1,分别用50、100、200 μg/ml予以刺激HUVEC,空白对照组采用RPMI-1640培养.6、12、24 h后分别行流式细胞仪和TUNEL法检测细胞凋亡率.Real time PCR和Western blot检测HUVEC中Fas和Caspase-3表达情况.结果 HSP患儿的IgA1(200 μg/ml)可诱导脐静脉内皮细胞发生凋亡,其凋亡率显著高于空白对照组[(14.77 ±2.23)％ vs.(2.25 ±0.77)％,P＜0.01]；健康对照的IgA1(200μg/ml)也可诱导脐静脉内皮细胞发生凋亡,凋亡率也显著高于空白对照组[(7.97±1.48)％ vs.(2.25±0.77)％,P＜0.01].HSP患儿的IgA1诱导HUVEC的凋亡呈浓度依赖及时间依赖性.而且HSP患儿的IgA1能显著促进HUVEC中Caspase-3及Fas的表达(P＜0.01).结论 HSP患儿体内IgA1可诱导HUVEC凋亡,其可能参与了HSP发生发展的过程.
目的 探討過敏性紫癜(HSP)患兒血清IgA1對臍靜脈內皮細胞(HUVEC)凋亡的影響.方法 培養人HUVEC,分為HSP組、正常對照組和空白對照組.親和層析法分離HSP患兒及正常兒童血清IgA1,分彆用50、100、200 μg/ml予以刺激HUVEC,空白對照組採用RPMI-1640培養.6、12、24 h後分彆行流式細胞儀和TUNEL法檢測細胞凋亡率.Real time PCR和Western blot檢測HUVEC中Fas和Caspase-3錶達情況.結果 HSP患兒的IgA1(200 μg/ml)可誘導臍靜脈內皮細胞髮生凋亡,其凋亡率顯著高于空白對照組[(14.77 ±2.23)％ vs.(2.25 ±0.77)％,P＜0.01]；健康對照的IgA1(200μg/ml)也可誘導臍靜脈內皮細胞髮生凋亡,凋亡率也顯著高于空白對照組[(7.97±1.48)％ vs.(2.25±0.77)％,P＜0.01].HSP患兒的IgA1誘導HUVEC的凋亡呈濃度依賴及時間依賴性.而且HSP患兒的IgA1能顯著促進HUVEC中Caspase-3及Fas的錶達(P＜0.01).結論 HSP患兒體內IgA1可誘導HUVEC凋亡,其可能參與瞭HSP髮生髮展的過程.
목적 탐토과민성자전(HSP)환인혈청IgA1대제정맥내피세포(HUVEC)조망적영향.방법 배양인HUVEC,분위HSP조、정상대조조화공백대조조.친화층석법분리HSP환인급정상인동혈청IgA1,분별용50、100、200 μg/ml여이자격HUVEC,공백대조조채용RPMI-1640배양.6、12、24 h후분별행류식세포의화TUNEL법검측세포조망솔.Real time PCR화Western blot검측HUVEC중Fas화Caspase-3표체정황.결과 HSP환인적IgA1(200 μg/ml)가유도제정맥내피세포발생조망,기조망솔현저고우공백대조조[(14.77 ±2.23)％ vs.(2.25 ±0.77)％,P＜0.01]；건강대조적IgA1(200μg/ml)야가유도제정맥내피세포발생조망,조망솔야현저고우공백대조조[(7.97±1.48)％ vs.(2.25±0.77)％,P＜0.01].HSP환인적IgA1유도HUVEC적조망정농도의뢰급시간의뢰성.이차HSP환인적IgA1능현저촉진HUVEC중Caspase-3급Fas적표체(P＜0.01).결론 HSP환인체내IgA1가유도HUVEC조망,기가능삼여료HSP발생발전적과정.
Objective To observe the effect of apoptosis of human umbilical vein endothelial cells (HUVEC) induced by IgA1 from Henoch-Sch(o)nlein purpura (HSP) patients.Method HUVEC were cultured in 3 different conditional media with IgA1 from HSP patients,normal healthy children and simply the cell culture medium.Serum IgA1 was purified by jacalin affinity chromatography,rates of apoptosis in HUVEC cells at different concentration and different times after incubation with IgA1 were determined by TUNEL method and flow cytometry.Real-time PCR and Western blot methods were used to detect the expression of caspase-3 and Fas,respectively.Result Apoptosis rate of HUVEC by IgA1 isolated from HSP patients were significantly higher than that of the blank control [(14.77 ± 2.23)％ vs.(2.25 ± 0.77)％,P ＜0.01] and the apoptosis rate of HUVEC induced by IgA1 from normal healthy children was higher than that of blank control [(7.97 ± 1.48) ％ vs.(2.25 ± 0.77) ％,P ＜ 0.01].The apoptosis rate of HUVEC induced by IgA1 from HSP was time and concentration-dependent.Moreover IgA1 isolated from HSP patients could significantly increase the caspase-3 and Fas expression (P ＜ 0.01).Conclusion The IgA1 from HSP patients could induce the apoptosis of HUVEC,which might be related to the progression of HSP.
