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The Effects of Thrombospondin 1 in Transforming Growth Factorβ1 Induce Rat Cardiac Fibroblasts

Author: ChenLing
Tutor: HuJianXin
School: Nanchang University
Course: Internal Medicine
Keywords: Thrombospondin Vascular endothelial growth factor Antisense oligonucleotides Rat cardiac fibroblasts
CLC: R542.21
Type: Master's thesis
Year: 2007
Downloads: 74
Quote: 0
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Objective: To investigate thrombospondin 1 (TSP-1) and vascular endothelial growth factor (VEGF) in rats induced by transforming growth factor-β1 (TGF-β1) myocardial fibroblast proliferation and collagen synthesis process, TSP- 1 and VEGF relations provide a theoretical basis for the prevention and treatment of myocardial fibrosis. Method: digestion differential paste the wall method of primary cultured myocardial ,3-4 cells into fiber cells used in the experiment, the cell morphology was observed under an inverted microscope, anti-fibronectin protein antibody staining cells were identified. The experiment was divided into five groups: (1) control group, did not add any inducer cultured for 24 hours. ② of TGF-β1 in a dose-dependent group, were added to 10ng/ml, 20ng/ml, 30ng/mlTGF-β1 cultured for 24 hours. ③ TGF-β1 time-dependent group, join the TGF-β1 were cultured for 12, 24 and 48 hours. ④ antisense oligonucleotide group, added TGF-β1 and TSP-1 antisense oligonucleotides and cultured for 24 hours. The the ⑤ sense oligonucleotide Group added TGF-β1 and TSP-1 sense oligonucleotide after 24 hours of incubation. The growth of the number of Hydroxyproline Determination of collagen content were detected by MTT cardiac fibroblasts (CFs). By semi-quantitative RT-PCR analysis of VEGFmRNA in TSP-1 mRNA expression. The VEGFmRNA relative expression and the TSP-1mRNA relative expression level of correlation analysis. Results: 1, myocardial fibroblast proliferation and collagen: group of TGF-β1-induced myocardial into fibroblasts the MTT reaction A value and collagen content were significantly higher than those in the control group (P lt; 0.01) compared with control. The MTT reaction of TSP-1 antisense oligonucleotide TGF-β1 group A value and collagen synthesis magnitude were significantly lower than the TGF-β1 group (P lt; 0.01). No significant difference between the TSP-1 sense oligonucleotide group with TGF-β1 group (P gt; 0.05). 2 mRNA expression: (1) VEGFmRNA expression: ① 10ng/mlTGF-β1 group and the culture of the 12-hour group VEGFmRNA relative expression compared to the control group significantly increased (P lt; 0.01) ② from 20,30 ng/mlTGF-β1 group and 24 hours, 48 ??hours the group VEGFmRNA the relative expression than that of the control group was significantly lower (P lt; 0.01). ③ TSP-1 antisense oligonucleotide group than 20ng/mlTGF-β1 group of VEGFmRNA the relative expression level was significantly higher (P lt; 0.01). ④ TSP-1 the sense oligonucleotide Group compared with 20ng/mlTGF-β1 groups of VEGFmRNA in relative expression levels, no significant changes (P gt; 0.05). (2) TSP-1mRNA expression the: ① 10ng/mlTGF-β1 group and culture of TSP-1mRNA in 12 hours group relative expression than that of the control group had no significant difference (P gt; 0.05). ② groups 20,30 ng/mlTGF-β1 and 24 hours and 48 hours groups of TSP-1mRNA the relative expression was significantly higher than that of the control group (P lt; 0.01). The ③ TSP-1 antisense oligonucleotide Group 20ng/ml TGF-β1 compare the TSP-1mRNA relative expression levels significantly decreased (P lt; 0.01). The ④ TSP-1 sense oligonucleotide Group over 20ng/mlTGF-β1 group relative expression level of TSP-1mRNA no significant change (P gt; 0.05). (3) Correlation analysis: correlation analysis showed that expression of the amount of time and concentration-dependent group VEGFmRNA of TGF-β1 and TSP-1mRNA r values ??of -0.537 and -0.714 (P lt; 0.01). VEGF and TSP-1 expression was negatively correlated. Conclusion: TGF-β1-induced rat cardiac fibroblast proliferation and collagen synthesis process elevated TSP-1 expression is induced by TGF-β1. 2.TSP-1 expression is inhibited VEGF expression. 3.TSP-1 antisense oligonucleotides can inhibit myocardial TGF-β1-induced fibroblast proliferation and collagen synthesis. 4.TSP-1 antisense oligonucleotide may have the role against myocardial fibrosis.

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CLC: > Medicine, health > Internal Medicine > Heart, blood vessels ( circulatory ) disease > Heart disease > Myocardial diseases > Myocarditis
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