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目的:观察TGF-β1对原代培养的新生大鼠心肌成型与Ⅳ胶原表达的影响。方法:取出生3~7d的SD大鼠心脏,利用酶消化法结合差速贴壁体外培养心肌成纤维细胞;分别用不同浓度的TGF-β1(TGF-β1=10 ng/mL、1ng/mL、0 ng/mL)干预心肌成纤维细胞,分别于干预后5h、24h,采用RT-PCR检测I型胶原和Ⅳ型胶原的表达。结果:1.用0ng/mL(对照组),1ng/mL和10ng/mL的TGF-β1干预心肌成纤维细胞5h,RT-PCR检测I型胶原/β-actin光密度值分别为0.61±0.02,0.73±0.03和0.86±0.04,光密度值随着TGF-β1浓度的增加而增加,各组比较,差异有显著性(P<0.01);干预心肌成纤维细胞24h后,RT-PCR检测I型胶原/β-actin光密度值分别为0.65±.03,0.91±0.02和0.98±0.02,光密度值随着TGF-β1浓度的增加而增加,各组比较,差异有显著性(P<0.01)。2.用0 ng/mL(对照组),1ng/mL and 10ng/mL的TGF-β1干预心肌成纤维细胞5h,RT-PCR检测Ⅳ型胶原/β-actin光密度值分别为0.58±0.06,0.71±0.02,0.87±0.02,光密度值随着TGF-β1浓度的增加而增加,各组比较,差异有显著性(P<0.01);干预心肌成纤维细胞24h后,RT-PCR检测Ⅳ型胶原/β-actin光密度值分别为0.62±0.01,0.83±0.05,0.96±0.02,光密度值随着TGFβ1浓度的增加而增加,各组比较,差异有显著性(P<0.01)。结论:TGFβ1能加强I型和Ⅳ型胶原的表达,在24小时内呈时间和剂量依赖性。
OBJECTIVE: To observe the effects of TGF-β1 on cardiac myogenic and collagen IV expression in primary cultured neonatal rats. Methods: Cardiac fibroblasts were isolated from SD rats of 3 ~ 7 days old and cultured in vitro by using enzyme digestion and differential adherent method. The fibroblasts were cultured with different concentrations of TGF-β1 (TGF-β1 = 10 ng / mL, 1 ng / mL , 0 ng / mL), respectively. The expressions of type I collagen and type IV collagen were detected by RT-PCR at 5 and 24 h after intervention respectively. Myocardial fibroblasts were treated with 0 ng / mL (control group), 1 ng / mL and 10 ng / mL of TGF-β1 for 5 h, and the optical density of type I collagen / β-actin was 0.61 ± 0.02 , 0.73 ± 0.03 and 0.86 ± 0.04, respectively. The optical density value increased with the increase of TGF-β1 concentration, the difference was significant (P <0.01); After intervention of myocardial fibroblasts for 24 h, The optical density of β-actin was 0.65 ± 0.03, 0.91 ± 0.02 and 0.98 ± 0.02, respectively. The optical density value increased with the increase of TGF-β1 concentration, the difference was significant (P <0.01) ). The myocardial fibroblasts were treated with 0 ng / mL (control group), 1 ng / mL and 10 ng / mL of TGF-β1 for 5 h, the optical density of type Ⅳ collagen / β-actin was 0.58 ± 0.06 by RT- 0.71 ± 0.02,0.87 ± 0.02, the optical density value increased with the increase of TGF-β1 concentration, the difference was significant (P <0.01); After intervention of fibroblasts for 24h, RT- The optical density of collagen / β-actin was 0.62 ± 0.01, 0.83 ± 0.05 and 0.96 ± 0.02, respectively. The optical density increased with the increase of TGFβ1 concentration. The difference was significant (P <0.01). Conclusion: TGFβ1 can enhance the expression of type I and type IV collagen in a time and dose-dependent manner within 24 hours.