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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 鄭景暉 | |
dc.contributor.author | Po-Shuen Lin | en |
dc.contributor.author | 林柏萱 | zh_TW |
dc.date.accessioned | 2021-06-17T02:46:12Z | - |
dc.date.available | 2018-09-13 | |
dc.date.copyright | 2017-09-13 | |
dc.date.issued | 2017 | |
dc.date.submitted | 2017-08-16 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/68996 | - |
dc.description.abstract | 實驗目的:轉型生長因子(Transforming growth factor-beta, TGF-β) 在發生蛀牙時,產生牙髓組織的修復與牙本質母細胞的分化與基質生成扮演重要的角色。TGF-β1經由許多不同的訊息傳導路徑來發揮它的效果,像是SMAD 路徑與 MAPK路徑。
本篇論文分兩部分來探討TGF-β1對牙髓組織修復與再生的影響。第一部分討論不同訊息傳導路徑對於牙髓細胞中TGF-β1在膠原蛋白(collagen),第三型基質金屬蛋白酶 (metalloproteinase-3, MMP-3) 與第一型組織抑制金屬蛋白酶 (tissue inhibitor metalloproteinase-1, TIMP-1)生成的影響。第二部分探討TGF-β1與第二型環氧酶(cyclooxygenase-2, COX-2)、前列腺素E2 (prostaglandin E2, PGE2)這兩種重要的發炎相關酵素的關係。並且進一步討論TGF-β1的不同訊息傳導路徑對於第二型環氧酶的影響。 實驗方法:使用TGF-β1對人類牙髓細胞做刺激,在某些實驗中則先加入U0126 (MEK/ERK抑制劑) ,SB431542 (ALK5/Smad2抑制劑) 等訊息傳導路徑的抑制劑做前處理。第一部分使用膠原蛋白定量測定(Sircol Collagen Assay) 測量牙髓細胞中膠原半白的量。培養液中的pro-collagen I, TIMP-1 and MMP-3的量以酵素免疫分析法(Enzyme-Linked ImmunoSorbent Assay, ELISA)來測定。第二部分以酵素免疫分析法(Enzyme-Linked ImmunoSorbent Assay, ELISA) 來測定PGE2的量。以反轉錄鏈聚合反應(RT-PCR)、西方點墨法(Western Blot Analysis)來測定COX-2 mRNA 與蛋白質的量。 實驗結果:TGF-β1 會促進牙髓細胞生成膠原蛋白,也會增加pro-collagen I 與 TIMP-1 的產生,但是對於MMP-3會有輕微的抑制傾向。 SB431542與 U0126這兩種訊息傳導路徑的抑制劑會抑制 TGF-β1所誘導的膠原蛋白與TIMP-1的產生。 TGF-β1 (1-10 ng/ml)會增加牙髓細胞內 COX-2與 PGE2的產生。 SB431542會抑制牙髓細胞內TGF-β1對於 COX-2的作用。而 U0126對於TGF-β1也有稍微抑制的現象。 結論:TGF-β1 會參與牙髓細胞受傷後產生的修復與再生過程,這個現象是經由ALK5/Smad2/3與MEK/ERK的訊息傳導路徑去刺激膠原蛋白與TIMP-1的生成。TGF-β1也透過這個訊息傳導路徑去增加COX-2 與PGE2的生成,這與牙髓細胞受傷後早期的發炎現象與之後的修復有關。 | zh_TW |
dc.description.abstract | Aim: TGF-β1 is an important growth factor which shown to influence odontoblast differentiation and matrix deposition in reactionary dentinogenesis during dental caries. TGF-β1 exerts its effects through many signaling pathways, such as SMADs and MAPKs.
In this study, there are two parts investigating effects of TGF-β1on pulpal repair and regeneration. In the first part, we investigated the differential signaling pathways responsible for effects of TGF-β1 on collagen turnover, metalloproteinase-3 (MMP-3) and tissue inhibitor metalloproteinase-1 (TIMP-1) production in human dental pulp cells. In the second part of this study, we investigated the relationships between TGF-β1, cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2), two important proinflammatory cytokines of pulpal inflammation in in human dental pulp cell and further clarify the differential signaling transduction pathways of TGF-β1 that influence COX-2 production. Materials and Methods: Pulp cells were exposed to TGF-β1 with/without pretreatment of SB431542 (an ALK5/Smad2/3 inhibitor) and U0126 (a MEK/ERK inhibitor). In the first part of this study, sircol collagen assay was used to measure cellular collagen content. Culture medium pro-collagen I, TIMP-1 and MMP-3 levels were determined by enzyme-linked immunosorbant assay (ELISA). In the second part of this study, ELISA was used for measurement of PGE2 levels. RT-PCR and western blot were used to determined COX-2 mRNA and protein, respectively. Results: TGF-β1 increased the collagen content, pro-collagen I and TIMP-1 production, but slightly decreased MMP-3 production of pulp cells. SB431542 and U0126 prevented the TGF-β1-induced increase of collagen content and TIMP-1 production of dental pulp cells. Exposure to TGF-β1 (1-10 ng/ml) increased the COX-2 level of cultured pulp cells. Exposure to TGF-β1 (0.1-10 ng/mL) significantly increased PGE2 level in dental pulp cells. Under the pretreatment of SB431542, the stimulatory effect of TGF-β1 on COX-2 level of pulp cells was inhibited. Similarly, U0126 also partly inhibited the TGF-β1-induced COX-2 expression. Conclusion: TGF-β1 may be involved in the healing/regeneration processes of dental pulp in response to injury by stimulation of collagen and TIMP-1 production. These events are associated with ALK5/Smad2/3 and MEK/ERK signaling. TGF-β1 increased the COX-2 and PGE2 level of cultured pulp cells. This effect was associated with ALK5/Smad2/3 and MEK/ERK pathways. These events are important in the early inflammation, repair and regeneration of dental pulp in response to injury. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T02:46:12Z (GMT). No. of bitstreams: 1 ntu-106-D98422002-1.pdf: 1765573 bytes, checksum: fd5008d8a997396c9a9b3f9f384bc1ba (MD5) Previous issue date: 2017 | en |
dc.description.tableofcontents | 中文摘要...5
Abstract...7 Part I...10 Chapter I Literature Review...10 1.1 Dental pulp and its repair...10 1.2 Transforming Growth Factor –β (TGF-β)...13 1.3 Receptors of TGF-β...15 1.4 ALK-1 & ALK5...17 1.5 ALK-3...17 1.6 Signaling transduction pathways of TGF-β...18 1.7 Matrix Metalloproteinases (MMPs) and Tissue Inhibitors of Metalloproteinases (TIMPs)...21 Chapter II The Purposes of the Study...23 Chapter III Materials and Methods...24 3.1 Materials...24 3.2 Culture of human dental pulp cells...24 3.3 Expression of various TGF-β receptor family members in dental pulp cells...25 3.4 Collagen content assay...26 3.5 Effect of TGF-β1 on pro-collagen I, TIMP-1 and MMP-3 secretion of pulp cells...28 3.6 Statistical analysis...28 Chapter IV Results...29 4.1 Expression of TGF-β receptor family members in dental pulp cells...29 4.2 Effect of TGF-β1 on the collagen content of dental pulp cells and its modulation by SB431542 and U0126...29 4.3 Effect of TGF-β1 on TIMP-1 level in the culture medium of dental pulp cells...30 4.4 Effect of TGF-β1 on MMP-3 level in the culture medium of dental pulp cells...30 Chpater V Discussion...31 Chapter VI Conclusion...37 Part II...38 Chapter I Literature Review...38 1.1 Biosynthesis of Prostaglandins, PGs...38 1.2 Cyclooxygenase...38 1.3 Signal transduction pathway of PGE2...39 1.4 Role of PGE2 in inflammation...40 1.5 Role of PGE2 in dental pulp and pulpal inflammation...41 1.6 Pulpal dentin complex and its repair in correlation with inflammation...42 Chapter II The Purposes of the Study...44 Chapter III Materials and Methods...45 3.1 Materials...45 3.2 Cell culture...45 3.3 Effect of TGF-β1 on PGE2 production of dental pulp cells: ELISA assay...46 3.4 Effect of TGF-β1 on COX-2 mRNA expression in dental pulp cells: RT-PCR...47 3.5 Effect of TGF-β1 on COX-2 protein expression: western blotting analysis...48 3.6 Statistical Analysis...50 Chapter IV Results...51 4.1 Effect of TGF-β1 on PGE2 production of dental pulp cells...51 4.2 Effect of TGF-β1 on COX-2 mRNA and protein expression in dental pulp cells...51 4.3 Effect of TGF-β1 on COX-2 expression in dental pulp cells and its modulation by SB431542 and U0126...51 Chpater V Discussion...53 Figures...59 Figure 1: Signal Transduction Pathway of TGF-ß Superfamily...59 Figure 2: The Smad Pathway of TGF-β Superfamily...60 Figure 3: The MEK/ERK Pathway of TGF-β Superfamily...61 Figure 4: The Hypothesis of This Study...62 Figure 5: Expression of TGF-β receptor family members in human dental pulp cells...63 Figure 6: Effect of TGF-β1 on the collagen content of cultured dental pulp cells...65 Figure 7: Effect of TGF-β1 on the pro-collagen I secretion of dental pulp cells...66 Figure 8: Effect of TGF-β1 on the TIMP-1 secretion of cultured dental pulp cells...68 Figure 9: Effect of TGF-β1 on the MMP-3 secretion of cultured dental pulp cells...71 Figure 10: Effect differential concentration of TGF-β1 on the PGE2 level in cultured dental pulp cells...72 Figure 11: Effect of different concentrations of TGF-β1 on the COX-2 mRNA expression in cultured dental pulp cells...73 Figure 12: Effect differential concentration of TGF-β1 on the COX-2 expression in cultured dental pulp cells...74 Figure 13: Effect differential concentration of TGF-β1 on the COX-2 expression and its modulation with SB431542 in cultured dental pulp cells...75 Figure 14: Effect differential concentration of TGF-β1 on the COX-2 expression and its modulation with U0126 in cultured dental pulp cells...76 References...77 | |
dc.language.iso | en | |
dc.title | TGF-β1對人類牙髓細胞生成膠原蛋白,前列腺素E2,第二型環氧酶的影響與其訊息傳導機制的探討 | zh_TW |
dc.title | Effect of TGF-beta1 on Collagen Turn-over, Prostaglandin E2, and Cyclooxygenase-2 in Human Dental Pulp Cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 105-2 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 高嘉澤,涂明君,張美姬,陳羿貞,張曉華 | |
dc.subject.keyword | 第五型激活素受體樣激?,膠原蛋白,第二型環氧?,牙髓組織,修復/再生,組織抑制金屬蛋白?,轉型生長因子,前列腺素,發炎, | zh_TW |
dc.subject.keyword | ALK5,Collagen,Cyclooxygenase-2,Dental pulp,Repair/Regeneration,Tissue inhibitor metalloproteinase,TGF-β1,Prostaglandin,Inflammation, | en |
dc.relation.page | 87 | |
dc.identifier.doi | 10.6342/NTU201703310 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2017-08-16 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 臨床牙醫學研究所 | zh_TW |
顯示於系所單位: | 臨床牙醫學研究所 |
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