論文名稱:

單純皰疹病毒感染對細胞分泌前列腺素E2 extracellular signal-regulated kinase(ERK)PKB/Akt訊息傳遞活化的影響

 

Effects of herpes simplex virus on production of prostaglandin E2 and activation of extracellular signal-regulated kinase and PKB/Akt

研究生:

謝佳妤  Jia-Yu Hsie

 

(以作者名查詢陽明大學館藏系統)

 

(以作者名查詢全國圖書書目資訊網)

指導教授:

洪善  Shan-Ling Hung

        學位類別:

碩士

        學校名稱:

國立陽明大學

系所名稱:

口腔生物研究所

            學號:

39317005

          學年度:

94

          語文別:

中文

          出版年:

95

關鍵字:

單純皰疹病毒  herpes simplex virus

 

訊息傳遞  cell signaling

 

ERK  extra-cellular regulated kinase

 

Akt  Akt

全文說明:

(本論文 20070829 對校內公開)

 

電子全文

        論文頁數:

66

摘要:

單純皰疹病毒(herpes simplex virus, HSV)是常見的致病微生物。成年人中約有60~90%都曾被感染。第一型單純疱疹病毒(HSV-1)通常造成口腔感染,而導致齦口炎(gingivostomatitis)及唇疱疹(herpes labialis)。細胞在正常生理狀態時會釋放各種物質來維持正常的生理狀態,在受到刺激時也會經由改變細胞內的訊息傳遞,而影響分泌物質的種類及含量。病毒感染會刺激細胞,使細胞的訊息傳遞改變、或是影響細胞分泌出的物質,能夠使細胞活化來執行更多作用以對抗病毒感染,或者是使病毒本身達到更有效的感染。目前研究指出多種病毒感染後會使p44/p42 MAP kinase(即extra-cellular regulated kinase ERK)活化,進而影響病毒基因表現,以及PKB/Akt活化會參與細胞的抗凋亡(anti-apoptosis)及細胞存活(cell survival),進而影響病毒存活。另外前列腺素E2 prostaglandin E2PGE2)會影響多種病毒複製,而環氧化酵素-2 cyclooxygenase-2COX-2)為產生PGE2的速率決定步驟酵素。本論文利用西方墨點法偵測HSV-1病毒感染後牙齦造纖維母細胞(gingival fibroblast GF)及人類子宮頸上皮細胞株(HeLa)其ERKAkt活化情形,並且使用偵測Akt受質的抗體觀察Akt下游因子的改變情形,次之利用反轉錄酶連鎖反應(reverse transcriptase-polymerase chain reaction RT-PCR)觀察HSV-1病毒感染後,細胞內COX-2 mRNA表現情形,最後使用酵素免疫分析方法 (EIA, Enzyme-Linked Immunoassay)分析被感染後的牙齦造纖維母細胞及HeLa細胞PGE2分泌量。研究結果顯示感染後的牙齦造纖維母細胞及HeLa細胞其ERKAkt皆會活化,並且Akt下游因子有四個分子大小不同的蛋白改變。而被HSV-1感染的牙齦造纖維母細胞,在感染後8小時,其COX-2 mRNA表現會增加。此外感染後的牙齦造纖維母細胞其PGE2分泌量有增加的趨勢,而HeLa細胞則無明顯變化。本論文研究結果能對於HSV-1病毒感染後的活化訊息傳遞路徑與PGE2的分泌有更深一步的了解,以期能了解HSV-1病毒感染如何影響組織健康。

 

Herpes simplex virus is a causative agent in many oral infection which leads to gingivostomatitis and herpes labialis. Sixty to ninty percent of adults have been infected with this virus. Cells release a variety of mediators to maintain the normal physiological status, Once stimulated, changes in intracellular signals may affect the types and amounts of secreted molecules. Viral infection may stimulate cells, influences cellular signaling pathways, or effects secretion. These events may activate the cells to fight against viral infection, or to facilitate the infection. Studies on several viruses have suggested that viral infection may activates cell events, including p44/p42 MAP kinaseextra-cellular regulated kinase, ERK, and affects viral gene expression. Moreover, activation of PKB/Akt plays a role in cellular anti-apoptosis and cell survival that may lead to viral survival. In addition, prostaglandin E2PGE2is involved in the replications of many viruses. Cyclooxygenase-2COX-2 is the rate-limiting enzyme in PGE2 production. In this thesis, Western blot analysis was used to detect activation of ERK and Akt in HSV-1-infected gingival fibroblast GF and human cervical carcinoma cell lineHeLa, anti-Akt substrate antibody was used to detect the changes in down stream factors of Akt. Reverse transcriptase-polymerase chain reactionRT-PCRwas used to detect COX-2 mRNA expression after HSV-1 infection. EIAenzyme-linked immunoassay was used to analyze the PGE2 production in the HSV-1-infected GF and HeLa cells. The results showed that both ERK and Akt were activated in infected-GF and HeLa cells. Four down stream factors of Akt with different molecular weight were affected. COX-2 mRNA expression was increased in GF after incubated with HSV-1 for 8 hours. Additionally, PGE2 secretion was increased in infected GF, but not affected infected HeLa cells. The findings in this thesis provide information of the signal pathway and secretion of PGE2 in HSV-1-infected cell, and help to understand HSV-1-infection may influence health of tissue.