行政院國家科學委員會專題研究計畫 成果報告
Heme oxygenase 1 (Hsp32) 在無機砷與其代謝產物對血管 內皮與平滑肌細胞增生之角色探討(2/2)
計畫類別: 整合型計畫
計畫編號: NSC93-2321-B-038-009-
執行期間: 93 年 08 月 01 日至 94 年 07 月 31 日 執行單位: 臺北醫學大學生藥學研究所
計畫主持人: 陳彥州 共同主持人: 沈杏娟 計畫參與人員: 吳金燕
報告類型: 完整報告
處理方式: 本計畫可公開查詢
中 華 民 國 94 年 10 月 3 日
Heme oxygenase 1 (Hsp32) 在無機砷與其代謝產物對血管內皮與平 滑肌細胞增生之角色探討(2/2)
計畫類別: 整合型計畫
計畫編號: NSC 93-2321-B-038-009
執行期間: 93 年 08 月 01 日 至 94 年 07 月 31 日 執行單位: 臺北醫學大學生藥學研究所
計畫主持人: 陳彥州
共同主持人: 沈杏娟 計畫參與人: 吳金燕
Results of the present project:
Fig.1 Both As+3 and MMA+3, but not As+5, DMA+3, MMA+5, DMA+5, induce apoptosis in embryonic fibroblasts.
Fig. 2 Both As+3 and MMA+3, but not As+5, DMA+3, MMA+5, DMA+5, induce HO-1 protein expression and HSP90 protein procession with reducing telomerase activity in embryonic fibroblasts.
Fig. 3. NAC and catalase prevention of As+3 and MMA+3-induced HO-1 and HSP90 protein procession with attenuation of telomerase reduction.
MTT (OD 600 nm)
0.0 0.2 0.4 0.6 0.8
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**
## ##
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C + NAC CAT + NAC CAT
As+3 MM A+3
M 1 pe rc en ta g e (% )
0 10 20 30 50 60
CON + NAC CAT + NAC CAT
As+3
MMA+3
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**
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Fig. 4 NAC prevention of As+3 and MMA+3-induced mitochondrial dysfunction in embryonic fibroblast.
Fig. 5 DPI prevention of As+3 and MMA+3-induced cytotoxicity via blocking telomerse activation.
Fig. 6 Inhibition of HSP90 activity potentiates As+3 and MMA+3-inhibited telomerase activity by TRAP assay.
M1 percentage (%)
0 10 20 30 40 50 60 70
CON + 10 20 + 10 20
As+3
NAC NAC
MMA+3
**
**
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A M
MTT assay (OD 600)
0.0 0.2 0.4 0.6 0.8 1.0
CON GA + GA + GA
As+3 MMA+3
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** **
**
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Fig. 7. Activation of JNKs involves in As+3 and MMA+3-induced cell death in embryonic fibroblasts.
In conclusion
We provide first evidences to indicated that different biological effects of arsenics in embryonic fibroblasts. Induction of HO-1 gene expression at both mRNA and protein level and HSP90 protein procession was identified in As+3 and MMA-3-treated cells.
Data of pharmacological study showed that activation of JNKs via inducing protein phosphorylation and HSP90 but not HO-1 protein might be involved in the action of As+3 and MMA+3. In conclusion, As+3 and MMA+3 are active in inducing cell death via blocking telomerase activity, and expression of HO-1 and HSP90 protein procession is detected.
M TT a ss ay ( O D 600 )
0.0 0.2 0.4 0.6 0.8
CON
**
**
** **
**
**
##
##
PDSB SPLY + PDSBSPLY+ PDSBSPLY
** ** **
**
As+3 MMA+3
MTT assay (OD 600)
0.0 0.2 0.4 0.6 0.8 1.0 1.2
CON +
As+3 (40 µM) MMA+3 (20 µM)
**
DFx SnPP Hb DFxSnPPHb + DFxSnPPHb
**
** ** ** ** ** **
*
MTT ass ay (OD 600 nm)
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7
CON pcDNA HO-1/A HO-1/B CON
CON
CON CON pcDNApcDNApcDNApcDNA HO-1/A
HO-1/A
HO-1/A
HO-1/A HO-1/B
HO-1/B
HO-1/B
HO-1/B
MMA+3 (20 µM) As+3
(80 µM)
MMA+3 (40 µM) As+3
(40 µM)
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