目的 观察干扰素α(IFN α)对血小板衍生生长因子-BB(PDGF-BB)刺激的大鼠肝星状细胞(HSC)分泌Ⅰ型胶原及转化生长因子β1(TGF β1)基因表达的影响,探讨IFN α抗肝纤维化的可能机制.方法 体外培养大鼠HSC系rHSC-99,分别用0、0.0125、0.0250、0.0500,0.1000、0.2000,0.4000 ng/ml IFN α,PDGF-BB干预和两者共同干预,用四甲基偶氮唑盐实验观察各组对HSC细胞活力的影响,采用逆转录聚合酶链反应方法测定各组对HSC细胞Ⅰ型胶原mRNA和TGF β1 mRNA表达的影响.结果 (1)HSC细胞活力(A值)PDGF-BB干预组为1.35±0.22,空白对照组为0.89±0.12,两组比较,F=16.311,P＜0.05,差异有统计学意义,说明PDGF-BB可提高HSC细胞活力.0.025,0.050、0.100、0.2000,0.400ng/ml IFN α加PDGF-BB共干预组,A值分别为0.84±0.18.0.45±0.15、0.26±0.01、0.33±0.07,0.30±0.06,较空白对照组明显降低,F=7.430,P＜0.05,差异有统计学意义,说明IFN α与PDGF-BB共同作用可抑制HSC细胞活力,且在0.025-0.100 ng/ml范围内随着IFN α浓度的增加其抑制作用越明显.(2)0.050、0.100、0.200ng/ml IFN α加PDGF-BB共干预各组Ⅰ型胶原mRNA相对表达值分别为0.94±0.19、0.61±0.12,0.52±0.02,空白对照组为1.41±0.01,共干预各组比空白对照组均明显降低,F=127.921,P＜0.05,差异有统计学意义.0.050、0.100,0.200ng/mlIFN α加PDGF-BB共干预组各组TGFβ1 mRNA相对表达值分别为1.18±0.06、1.15±0.10、1.39±0.04,空白对照组为1.62±0.12,共干预各组比空白对照组均明显降低,F=82.115,P＜0.05,差异有统计学意义,说明IFN α与PDGF-BB共同作用对HSC细胞Ⅰ型胶原、TGFβ1基因的表达有抑制作用,且随着浓度的增加其抑制作用越明显.结论 IFN α对PDGF-BB诱导的HSC细胞活力及Ⅰ型胶原、TGF β1基因的表达有抑制作用,且随着浓度的增加其抑制作用越明显.这可能是IFN α发挥抗肝纤维化作用的途径之一.
目的 觀察榦擾素α(IFN α)對血小闆衍生生長因子-BB(PDGF-BB)刺激的大鼠肝星狀細胞(HSC)分泌Ⅰ型膠原及轉化生長因子β1(TGF β1)基因錶達的影響,探討IFN α抗肝纖維化的可能機製.方法 體外培養大鼠HSC繫rHSC-99,分彆用0、0.0125、0.0250、0.0500,0.1000、0.2000,0.4000 ng/ml IFN α,PDGF-BB榦預和兩者共同榦預,用四甲基偶氮唑鹽實驗觀察各組對HSC細胞活力的影響,採用逆轉錄聚閤酶鏈反應方法測定各組對HSC細胞Ⅰ型膠原mRNA和TGF β1 mRNA錶達的影響.結果 (1)HSC細胞活力(A值)PDGF-BB榦預組為1.35±0.22,空白對照組為0.89±0.12,兩組比較,F=16.311,P＜0.05,差異有統計學意義,說明PDGF-BB可提高HSC細胞活力.0.025,0.050、0.100、0.2000,0.400ng/ml IFN α加PDGF-BB共榦預組,A值分彆為0.84±0.18.0.45±0.15、0.26±0.01、0.33±0.07,0.30±0.06,較空白對照組明顯降低,F=7.430,P＜0.05,差異有統計學意義,說明IFN α與PDGF-BB共同作用可抑製HSC細胞活力,且在0.025-0.100 ng/ml範圍內隨著IFN α濃度的增加其抑製作用越明顯.(2)0.050、0.100、0.200ng/ml IFN α加PDGF-BB共榦預各組Ⅰ型膠原mRNA相對錶達值分彆為0.94±0.19、0.61±0.12,0.52±0.02,空白對照組為1.41±0.01,共榦預各組比空白對照組均明顯降低,F=127.921,P＜0.05,差異有統計學意義.0.050、0.100,0.200ng/mlIFN α加PDGF-BB共榦預組各組TGFβ1 mRNA相對錶達值分彆為1.18±0.06、1.15±0.10、1.39±0.04,空白對照組為1.62±0.12,共榦預各組比空白對照組均明顯降低,F=82.115,P＜0.05,差異有統計學意義,說明IFN α與PDGF-BB共同作用對HSC細胞Ⅰ型膠原、TGFβ1基因的錶達有抑製作用,且隨著濃度的增加其抑製作用越明顯.結論 IFN α對PDGF-BB誘導的HSC細胞活力及Ⅰ型膠原、TGF β1基因的錶達有抑製作用,且隨著濃度的增加其抑製作用越明顯.這可能是IFN α髮揮抗肝纖維化作用的途徑之一.
목적 관찰간우소α(IFN α)대혈소판연생생장인자-BB(PDGF-BB)자격적대서간성상세포(HSC)분비Ⅰ형효원급전화생장인자β1(TGF β1)기인표체적영향,탐토IFN α항간섬유화적가능궤제.방법 체외배양대서HSC계rHSC-99,분별용0、0.0125、0.0250、0.0500,0.1000、0.2000,0.4000 ng/ml IFN α,PDGF-BB간예화량자공동간예,용사갑기우담서염실험관찰각조대HSC세포활력적영향,채용역전록취합매련반응방법측정각조대HSC세포Ⅰ형효원mRNA화TGF β1 mRNA표체적영향.결과 (1)HSC세포활력(A치)PDGF-BB간예조위1.35±0.22,공백대조조위0.89±0.12,량조비교,F=16.311,P＜0.05,차이유통계학의의,설명PDGF-BB가제고HSC세포활력.0.025,0.050、0.100、0.2000,0.400ng/ml IFN α가PDGF-BB공간예조,A치분별위0.84±0.18.0.45±0.15、0.26±0.01、0.33±0.07,0.30±0.06,교공백대조조명현강저,F=7.430,P＜0.05,차이유통계학의의,설명IFN α여PDGF-BB공동작용가억제HSC세포활력,차재0.025-0.100 ng/ml범위내수착IFN α농도적증가기억제작용월명현.(2)0.050、0.100、0.200ng/ml IFN α가PDGF-BB공간예각조Ⅰ형효원mRNA상대표체치분별위0.94±0.19、0.61±0.12,0.52±0.02,공백대조조위1.41±0.01,공간예각조비공백대조조균명현강저,F=127.921,P＜0.05,차이유통계학의의.0.050、0.100,0.200ng/mlIFN α가PDGF-BB공간예조각조TGFβ1 mRNA상대표체치분별위1.18±0.06、1.15±0.10、1.39±0.04,공백대조조위1.62±0.12,공간예각조비공백대조조균명현강저,F=82.115,P＜0.05,차이유통계학의의,설명IFN α여PDGF-BB공동작용대HSC세포Ⅰ형효원、TGFβ1기인적표체유억제작용,차수착농도적증가기억제작용월명현.결론 IFN α대PDGF-BB유도적HSC세포활력급Ⅰ형효원、TGF β1기인적표체유억제작용,차수착농도적증가기억제작용월명현.저가능시IFN α발휘항간섬유화작용적도경지일.
Objective To investigate the effect of IFN alpha on the expressions of Collagen land TGF|(A) 1 jn hepatic stellate eell activated by PDGF-BB.Methods Hepatic stellate cells(rHSC-99)treated with IFN alpha of different concentration(0,0.0125,0.025,0.050,0.100,0.200,0.400 ng/m1).The cell viability of HSC was measured by MTT.The leveis of Col-I mRNA and TGF β1 mRNA were measured by the quantitative reverse-transcription polymerase chain reaction(RT-PCR).Results (1)When HSC was exposed in PDGF-BB,the cell viability of HSC (1.35±0.22) was higher than that of the control group (0.890±0.12) (F=16.311,P＜0.05),indicating that PDGF-BB can promote the cell viability of HSC. When HSC was exposed to both PDGF-BB and different concentration of IFN alpha (0.025,0.05,0.1,0.2,0.4 ng/ml),the cell viability of HSC (0.840±0.18,0.450±0.15,0.260±0.01,0.330±0.07,0.30±0.06) were lower than that of the control group (0.890±0.12) (F=7.430,P＜0.05),indicating that the cell viability of HSC was inhibited when HSC was exposed to both PDGF-BB and different concentrations of IFN alpha. Furthermore,within the range of 0.025 ng/ml to 0.1 ng/ml,the effect of IFN alpha was dosedependant. (2). The relative expression values of Col-I mRNA in different groups of (0.05,0.1,0.2 ng/ml)IFN alpha +PDGF-BB are (0.940±0.19,0.610±0.12,0.520±0.02),which were lower than those in the control group (1.410±0.01) (F=127.921,P＜0.05). The relative expression values of TGF β1 mRNA in different groups of (0.05,0.1,0.2 ng/ml) IFN alpha +PDGF-BB are (1.180±0.06,1.150±0.10,1.390±0.04),again were lower than those in the control group (1.620±0.12) (F=82.115,P＜0.05). These results indicated that the expression of Col-I mRNA and TGF β1 mRNA was remarkably inhibited when HSC was exposed in both PDGF-BB and IFN alpha. Conclusion The cell viability of HSC and the expression of Col-I mRNA and TGFβI mRNA is remarkably inhibited when HSC is exposed in both PDGF-BB and IFN alpha,and the inhibition is dose-dependant.
