中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
11期
2490-2493
,共4页
谭维军%程真顺%谭秋月%叶燕青%江平
譚維軍%程真順%譚鞦月%葉燕青%江平
담유군%정진순%담추월%협연청%강평
碱性成纤维细胞生长因子%反义寡核苷酸%肺间质纤维化%转化生长因子-β1-Smad信号通路
堿性成纖維細胞生長因子%反義寡覈苷痠%肺間質纖維化%轉化生長因子-β1-Smad信號通路
감성성섬유세포생장인자%반의과핵감산%폐간질섬유화%전화생장인자-β1-Smad신호통로
Basic fibroblast growth factor%Antisense oligodeoxynucleotide%Pulmonary fibrosis%Transforming growth factor-β1-Smad
目的 观察碱性成纤维细胞生长因子(bFGF)反义寡核苷酸对大鼠肺纤维化的影响.方法 将30只大鼠随机分为正常对照组、肺纤维化组、空载体转染组、反义bFGF转染组、正义bFGF转染组,每组6只.造模处理后,计算各组大鼠肺系数,比较肺组织纤维化程度,酶联免疫吸附试验(ELISA)检测血清和肺泡灌洗液bFGF含量及肺匀浆羟脯氨酸含量,逆转录-聚合酶链反应(RT-PCR)检测肺匀浆中bFGF、α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(Collagen Ⅰ)、转化生长因子-β1(TGF-β1)、结缔组织生长因子(CTGF)、Smad3和Smad7的mRNA表达.结果 对照组、肺纤维化组、空载体转染组、反义bFGF组、正义bFGF组大鼠肺系数值分别为:(7.5±1.5)％、(1O.8±1.4)％、(11.0±1.4)％、(8.8±1.7)％、(12.5±1.6)％;肺间质纤维化分级为:0、2.38±0.52、2.41±0.48、1.62±0.45、2.82±0.54;反义bFGF转染组肺系数明显降低(P＜0.05),正义bFGF转染组肺系数明显增高(P＜0.05);反义bFGF转染组肺间质纤维化明显减轻(P＜0.05),正义bFGF转染组明显加重(P＜0.05);反义bFGF转染组血清、肺泡灌洗液中bFGF含量及肺匀浆中羟脯氨酸含量明显降低(P＜0.05),正义bFGF转染组含量明显增高(P＜0.05);反义bFGF转染组肺匀浆中α-SMA、Collagen Ⅰ、TGF-β1、CTGF及Smad3 mRNA的表达水平明显降低(P＜0.05),正义bFGF转染组表达明显增高(P＜0.05),Smad7 mRNA的表达刚好相反.结论 bFGF反义寡核苷酸可以减轻博莱霉素诱导的大鼠肺纤维化,可能机制是bFGF反义寡核苷酸下调了TGF-β1 Smad信号通路.
目的 觀察堿性成纖維細胞生長因子(bFGF)反義寡覈苷痠對大鼠肺纖維化的影響.方法 將30隻大鼠隨機分為正常對照組、肺纖維化組、空載體轉染組、反義bFGF轉染組、正義bFGF轉染組,每組6隻.造模處理後,計算各組大鼠肺繫數,比較肺組織纖維化程度,酶聯免疫吸附試驗(ELISA)檢測血清和肺泡灌洗液bFGF含量及肺勻漿羥脯氨痠含量,逆轉錄-聚閤酶鏈反應(RT-PCR)檢測肺勻漿中bFGF、α-平滑肌肌動蛋白(α-SMA)、Ⅰ型膠原(Collagen Ⅰ)、轉化生長因子-β1(TGF-β1)、結締組織生長因子(CTGF)、Smad3和Smad7的mRNA錶達.結果 對照組、肺纖維化組、空載體轉染組、反義bFGF組、正義bFGF組大鼠肺繫數值分彆為:(7.5±1.5)％、(1O.8±1.4)％、(11.0±1.4)％、(8.8±1.7)％、(12.5±1.6)％;肺間質纖維化分級為:0、2.38±0.52、2.41±0.48、1.62±0.45、2.82±0.54;反義bFGF轉染組肺繫數明顯降低(P＜0.05),正義bFGF轉染組肺繫數明顯增高(P＜0.05);反義bFGF轉染組肺間質纖維化明顯減輕(P＜0.05),正義bFGF轉染組明顯加重(P＜0.05);反義bFGF轉染組血清、肺泡灌洗液中bFGF含量及肺勻漿中羥脯氨痠含量明顯降低(P＜0.05),正義bFGF轉染組含量明顯增高(P＜0.05);反義bFGF轉染組肺勻漿中α-SMA、Collagen Ⅰ、TGF-β1、CTGF及Smad3 mRNA的錶達水平明顯降低(P＜0.05),正義bFGF轉染組錶達明顯增高(P＜0.05),Smad7 mRNA的錶達剛好相反.結論 bFGF反義寡覈苷痠可以減輕博萊黴素誘導的大鼠肺纖維化,可能機製是bFGF反義寡覈苷痠下調瞭TGF-β1 Smad信號通路.
목적 관찰감성성섬유세포생장인자(bFGF)반의과핵감산대대서폐섬유화적영향.방법 장30지대서수궤분위정상대조조、폐섬유화조、공재체전염조、반의bFGF전염조、정의bFGF전염조,매조6지.조모처리후,계산각조대서폐계수,비교폐조직섬유화정도,매련면역흡부시험(ELISA)검측혈청화폐포관세액bFGF함량급폐균장간포안산함량,역전록-취합매련반응(RT-PCR)검측폐균장중bFGF、α-평활기기동단백(α-SMA)、Ⅰ형효원(Collagen Ⅰ)、전화생장인자-β1(TGF-β1)、결체조직생장인자(CTGF)、Smad3화Smad7적mRNA표체.결과 대조조、폐섬유화조、공재체전염조、반의bFGF조、정의bFGF조대서폐계수치분별위:(7.5±1.5)％、(1O.8±1.4)％、(11.0±1.4)％、(8.8±1.7)％、(12.5±1.6)％;폐간질섬유화분급위:0、2.38±0.52、2.41±0.48、1.62±0.45、2.82±0.54;반의bFGF전염조폐계수명현강저(P＜0.05),정의bFGF전염조폐계수명현증고(P＜0.05);반의bFGF전염조폐간질섬유화명현감경(P＜0.05),정의bFGF전염조명현가중(P＜0.05);반의bFGF전염조혈청、폐포관세액중bFGF함량급폐균장중간포안산함량명현강저(P＜0.05),정의bFGF전염조함량명현증고(P＜0.05);반의bFGF전염조폐균장중α-SMA、Collagen Ⅰ、TGF-β1、CTGF급Smad3 mRNA적표체수평명현강저(P＜0.05),정의bFGF전염조표체명현증고(P＜0.05),Smad7 mRNA적표체강호상반.결론 bFGF반의과핵감산가이감경박래매소유도적대서폐섬유화,가능궤제시bFGF반의과핵감산하조료TGF-β1 Smad신호통로.
Objective To observe the effect of antisense oligodeoxynucleotide (AODN) of basic fibroblast growth factor (bFGF) on the rat models of pulmonary fibrosis.Methods Thirty rats were divided into five groups (6 rats of each group):control group,pulmonary fibrosis group,empty vector transfection group,AODN group and RODN group.Lung coefficient was counted,pathologic changes in the lung tissue were observated to determine degree of pulmonary fibrosis,enzyme linked immunosorbent assay (ELISA) was used to determine the concentrations of hydroxyproline in lung tissue homogenate and the expression of bFGF in the serum and bronchoalveolar lavage fluid,and reverse transcription-polymerase chain reaction (RT-PCR) was employed to detect the mRNA expression of bFGF,α-smooth muscle actin (α-SMA),type Ⅰ collagen,transforming growth factor-β1 (TGF-β1),connective tissue growth factor (CTGF),Smad3 and Smad7 in lung tissue homogenate.Results In control group,pulmonary fibrosis gToup,empty vector transfection group,AODN group and RODN group,the lung coefficient was 7.5 ± 1.5,10.8 ± 1.4,11.0 ± 1.4,8.8 ± 1.7 and 12.5 ± 1.6 respectively,the degree of pulmonary fibrosis was 0,2.38 ± 0.52,2.41 ± 0.48,1.62 ± 0.45 and 2.82 ±0.54 respectively,the synthesis of bFGF was significantly decreased in AODN group and significantly increased in RODN group (P ＜ 0.05),the concentration of HYP was markedly decreased in AODN group and markedly increased in RODN group (P ＜ 0.05),the mRNA expression of bFGF,α-SMA,type Ⅰ collagen,TGF-β1,CTGF and Smad3 was markedly decreased in AODN group and markedly increased RODN group (P ＜ 0.05),and that of Smad7 was markedly increased in AODN group and markedly decreased in RODN group.Conclusion:These results demonstrate that AODN of bFGF can inhibit the development of pulmonary fibrosis induced by Bleomycyn in rats possibly by down-regulating the TGF-β1-Smad signaling pathway.
