過去已經發現在急性劇烈運動下，體內會產生大量的熱使得體溫升高和產生氧化壓力。這兩種壓力會造成細胞內的熱休克轉錄因子 (heat shock factor-1; HSF-1) 改變下游基因的轉錄效率。根據微陣列晶片結果發現在急性劇烈腳踏車運動後，很多白血球的基因會發生改變。因此我們透過生物資訊的網路平台 (TESS和Ensembl) 探討在93條急性運動後表現會上升的基因，其在啟動子的區段又含有 HSF-1 的結合位 (hsf-1 binding element; HSE) 的四條基因的表現。首先，我們探討在急性劇烈運動後是否可以藉由活化 HSF-1 進而去調控下游基因。我們抽取受試者運動前後以及抽出後單純透過加熱處理的靜脈血，分離出靜脈血中的白血球後，利用免疫染色發現，只有在加熱組的白血球細胞核中發現 HSF-1 有聚集的現象，而急性劇烈運動後則沒有這種現象發生。接著，利用 real time PCR 確認啟動子區段具有 HSE 的微陣列基因表現，結果發現在運動後 HSPA1B 和 CST7 基因 的mRNA 表現量顯著上升，而 PTGDR 和 CD244 則沒有顯著性改變。最後，利用 luciferase assay 分析這些基因的啟動子活性則發現，HSPA1B 的啟動子活性在加熱後第12和24小時，分別上升約2.4倍和20%。，PTGDR 和 CD244 的啟動子活性僅在加熱後第12小時有些微上升。因此，我們發現只有加熱處理可以促進HSF-1進入細胞核中；另外，急性劇烈運動使其下游基因HSPA1B 和 CST7的表現量上升。而HSPA1B基因的表現量增加則可能和其啟動子活化有很大的關聯。

It is known that acute severe exercise induces heat production and oxidative stress. These changes may activate heat shock factor-1 (HSF-1) and alter the transcriptional rate of downstream genes. The results of Affymetrix microarray by using human leukocytes demonstrated that the expression of many genes was changed after acute severe ergometer exercise. Using bioinformatic online platforms, such as TESS and Ensembl, we found that the promoter regions of 8 out of 93 severe exercise-upregulated genes might have hsf-1 binding element (HSE). To investigate whether acute severe exercise activates the transcription factor, hsf-1, and hence regulates these genes in leukocytes, male subjects were subjected to a single bout of severe exercise until exhaustion. Venous blood samples were collected before and after exercise, and collected only for heat treatment. The immunocytochemistry results found that HSF-1 just translocated into leukocytes nucleus after heat treatment but not after acute severe exercise. In addition, Real-time PCR was used to detect the mRNA levels of the genes which are upregulated by acute exercise and may have HSE in their promoter regions. Our results confirmed that acute severe exercise induces expression of two genes, HSPA1B and CST7, but doesn’t change the expression of PTGDR and CD244. Furthermore, luciferase assay was used to investigate whether the gene upregulation is related to their HSE promoter activities. The results showed that heat shock treatment only induced about 2.4-fold and 20% increase of HSPA1B promoter activity in 12 and 24 hours after heat shock, and PTGDR and CD244 promoter activities were slightly increased only in 12 hours after heat sock. In conclusion, we found that only heat shock treatment can induce HSF-1 to translocate into nucleus. However, acute severe exercise increased the mRNA expressions of its downstream genes, HSPA1B and CST7 and the upregulation of HSPA1B gene may be related to the enhancement of its promoter activity.

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