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The Effects on the Proliferation Apoptosis and Collagen Synthesis of Human Embryo Fibroblast

Author: ZhangXiaoFei
Tutor: WangChangMing
School: Guilin Medical College,
Course: Internal Medicine
Keywords: artesunate embryonic lung fibroblasts procollagen apoptosis Bcl-2 mRNA Bax mRNA Survivin mRNA RT-PCR
CLC: R285
Type: Master's thesis
Year: 2010
Downloads: 27
Quote: 0
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Abstract


Objective To investigate the effects and mechanism of Artesunate on theproliferation and apoptosis of human embryo fibroblast, To investigate theeffects of Artesunate on the collagen synthesis of human embryo fibroblastHFL-I.Method Human embryo fibroblasts were incubated with 0、1、10、100mg/L，at a scheduled time，the proliferation activity of the human embryofibroblasts was determined by CCK-8 assay，the morphology of apoptosis wasobserved by Hochst33258 staining. the cellcycle phase and apoptosis ratio wereanalyzed by flow cytometry( FCM), Human embryonic lung fibroblasts (HELF)cell lines HFL-I were cultured to log phase in vitro and divided into 3 observedgroups and control group，the former 3 groups were treated with artesunate atthe mass concentration of 1、10、100mg/L，the control group were givenequivalent DMEM，24h later，the cells was collected and total RNA wasextracted，the expression of Bcl-2 mRNA，Bax mRNA and Survivin mRNA wasdetected by RT-PCR, Human embryo fibroblasts were incubated with 0、1、10、100mg/L，the collegan synthesis was observed by sirius red staining，theexpression of type I procollagen and type III procollagen mRNA was detected by RT-PCR.Results Compared with the normal control Artesunate significantlyinhibited HFL-I proliferation and restrained collagen synthesis in a dose-andtime-dependent manner (P<0. 05). Flow cytometry showed that，the number ofHFL-I was increased in the G0/G1 phase but decreased in the S phase，and thecells were arrested in the G0/G1 phase. Artesunate significantly inhibitsproliferation of human embryo fibroblast，makes it arrest in G0/G1 phase，Apoptosis rate was increased, Compared with the control group，the expressionof Bcl-2 mRNA and Survivin mRNA decreased significantly and the expressionof Bax mRNA was Contrary, Compared with the normal control，Artesunatesignificantly restrained collagen synthesis in a dose-dependent manner (P<0.05)，the expression of type I procollagen and type III procollagen mRNAdecreased significantly.Conclusion Artesunate can inhibit human embryonic lung fibroblastproliferation，promoting their apoptosis，and probable mechanism relate withinhibiting HFL-I proliferation and inducing HFL-I apoptosis throughdownregulating the expression of Bcl-2 mRNA and Survivin mRNA butupregulating the expression of Bax mRNA. Artesunate reduces the secretion ofcollagen probable mechanism relate with reduced the expression of type Iprocollagen and type III procollagen mRNA，those indicate that artesunate maycontrol pulmonary fibrosis.

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