北京医学
北京醫學
북경의학
BEIJING MEDICAL JOURNAL
2010年
4期
270-274
,共5页
上皮-间充质细胞转化%红细胞生成素%转化生长因子-β1%血管内皮生长因子%E钙黏蛋白α-平滑肌肌动蛋白
上皮-間充質細胞轉化%紅細胞生成素%轉化生長因子-β1%血管內皮生長因子%E鈣黏蛋白α-平滑肌肌動蛋白
상피-간충질세포전화%홍세포생성소%전화생장인자-β1%혈관내피생장인자%E개점단백α-평활기기동단백
Epithelial-mesenchymal transition(EMT)%Erythropoietin%Transform growth factor-β1 (TGF-β1)Vascular endothelial growth factor(VEGF)%E-cadherin protein%α-smooth muscle actin (α-SMA)
目的 探讨红细胞生成素(rHuEPO)对人肾小管上皮细胞(HKC)上皮-间充质细胞转化(EMT)的作用及其与该细胞血管内皮生长因子(VEGF)表达变化的关系.方法 体外培养HKC,以未处理的HKC为阴性对照,以8ng/ml的转化生长因子-β1(TGF-β1)处理的HKC为阳性对照,以不同浓度(0.1、1.0、10、50、100 IU/ml)的rHuEPO与8ng/ml的TGF-β1共同处理HKC,或在不同时间(-24、0、12、24、36 h)用同一浓度rHuEPO(100 IU/ml)对8ng/ml的TGF-β1处理的HKC进行干预.应用RT-PCR方法检测α-平滑肌肌动蛋白(α-SMA)和VEGFmRNA转录水平,应用免疫印迹方法检测α-SMA、E钙黏蛋白以及VEGF的蛋白表达水平.结果 rHuEPO以剂量和时间依赖的方式抑制TGF-β1诱导的HKC α-SMA mRNA和蛋白的表达(P＜0.05或P＜0.01),保护性上调TGF-β1抑制的HKC细胞E钙黏蛋白的表达(P＜0.05或P＜0.01);同时上调TGF-β1抑制的HKC细胞VEGF mRNA和蛋白的表达(P＜0.05或P＜0.01).结论 rHuEPO对TGF-β1诱导的EMT具有一定的抑制作用,并具剂量一时间依赖性;该作用可能与rHuEPO上调肾小管上皮细胞VEGF的表达有关.
目的 探討紅細胞生成素(rHuEPO)對人腎小管上皮細胞(HKC)上皮-間充質細胞轉化(EMT)的作用及其與該細胞血管內皮生長因子(VEGF)錶達變化的關繫.方法 體外培養HKC,以未處理的HKC為陰性對照,以8ng/ml的轉化生長因子-β1(TGF-β1)處理的HKC為暘性對照,以不同濃度(0.1、1.0、10、50、100 IU/ml)的rHuEPO與8ng/ml的TGF-β1共同處理HKC,或在不同時間(-24、0、12、24、36 h)用同一濃度rHuEPO(100 IU/ml)對8ng/ml的TGF-β1處理的HKC進行榦預.應用RT-PCR方法檢測α-平滑肌肌動蛋白(α-SMA)和VEGFmRNA轉錄水平,應用免疫印跡方法檢測α-SMA、E鈣黏蛋白以及VEGF的蛋白錶達水平.結果 rHuEPO以劑量和時間依賴的方式抑製TGF-β1誘導的HKC α-SMA mRNA和蛋白的錶達(P＜0.05或P＜0.01),保護性上調TGF-β1抑製的HKC細胞E鈣黏蛋白的錶達(P＜0.05或P＜0.01);同時上調TGF-β1抑製的HKC細胞VEGF mRNA和蛋白的錶達(P＜0.05或P＜0.01).結論 rHuEPO對TGF-β1誘導的EMT具有一定的抑製作用,併具劑量一時間依賴性;該作用可能與rHuEPO上調腎小管上皮細胞VEGF的錶達有關.
목적 탐토홍세포생성소(rHuEPO)대인신소관상피세포(HKC)상피-간충질세포전화(EMT)적작용급기여해세포혈관내피생장인자(VEGF)표체변화적관계.방법 체외배양HKC,이미처리적HKC위음성대조,이8ng/ml적전화생장인자-β1(TGF-β1)처리적HKC위양성대조,이불동농도(0.1、1.0、10、50、100 IU/ml)적rHuEPO여8ng/ml적TGF-β1공동처리HKC,혹재불동시간(-24、0、12、24、36 h)용동일농도rHuEPO(100 IU/ml)대8ng/ml적TGF-β1처리적HKC진행간예.응용RT-PCR방법검측α-평활기기동단백(α-SMA)화VEGFmRNA전록수평,응용면역인적방법검측α-SMA、E개점단백이급VEGF적단백표체수평.결과 rHuEPO이제량화시간의뢰적방식억제TGF-β1유도적HKC α-SMA mRNA화단백적표체(P＜0.05혹P＜0.01),보호성상조TGF-β1억제적HKC세포E개점단백적표체(P＜0.05혹P＜0.01);동시상조TGF-β1억제적HKC세포VEGF mRNA화단백적표체(P＜0.05혹P＜0.01).결론 rHuEPO대TGF-β1유도적EMT구유일정적억제작용,병구제량일시간의뢰성;해작용가능여rHuEPO상조신소관상피세포VEGF적표체유관.
Objective To examine the effect of rHuEPO on epithelial-mesenehymal transition(EMT) of human renal proximal tubular epithelail cells (HKC) cultured in vitro and to elucidate the relationship between EMT and vascular endothelial growth factor (VEGF) expression of the cells. Methods HKC cells were divided into three groups, including negative control (Untreated HKC), positive control (treated with TGF-β1, 8 ng/mL ) and HKC co-treated with rHuEPO (0.1,1.0,10,50,100 IU/ml) and TGF-β1 ( 8 ng/mL). HKC cells were incubated with the same concentration of rHuEPO (100 IU/ml) for variouse periods (-24h,0h,12h,24h,36h,48h). Western blot and RT-PCR were used to evaluate the mRNA and protein levels of α-SMA, E-cadherin and VEGF. Results The VEGF mRNA , VEGF and E-cadherin protein expressions significantly increased (P＜0.05或P＜0.01), while the α-SMA mRNA and protein expressions significantly decreased (P＜0.05或P＜0.01) in HKC cells concomitantly treated with rHuEPO and TGF-β1 when comparing with the positive control. The expressions of α-SMA mRNA and α-SMA protein were significantly decreased(P＜0.05或P＜0.01)while the expressions of VEGF mRNA, VEGF and E-eadherin protein were significantly increased (P＜0.05或P＜0.01)in rHuEPO (100 IU/mL) treated for various periods (-24h,Oh,12h,24h,36h,48h) when compared with the TGF-β1(8 ng/mL)treated alone. Conclusions rHuEPO may inhibit TGF-β1 induced EMT of cultured HKC cells in dose-and time-dependent manner. This effect is probably related to up-regulated expression of VEGF induced by rHuEPO.
