請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/80863完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王兆麟(Jaw-Lin Wang) | |
| dc.contributor.author | Yueh-Chun Huang | en |
| dc.contributor.author | 黃粵軍 | zh_TW |
| dc.date.accessioned | 2022-11-24T03:19:33Z | - |
| dc.date.available | 2021-11-08 | |
| dc.date.available | 2022-11-24T03:19:33Z | - |
| dc.date.copyright | 2021-11-08 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-10-04 | |
| dc.identifier.citation | 1. Ethgen O, Beaudart C, Buckinx F, Bruyere O, Reginster JY. The Future Prevalence of Sarcopenia in Europe: A Claim for Public Health Action. Calcif Tissue Int 2017; 100:229-34. 2. Musar, #xf2, and A. , The Basis of Muscle Regeneration. Advances in Biology, 2014. 2014: p. 16. 3. Burattini, S., et al., C2C12 murine myoblasts as a model of skeletal muscle development: morpho-functional characterization. European Journal of Histochemistry, 2004. 48(3): p. 223-233. 4. McMahon, D.K., et al., C2C12 cells - biophysical, biochemical, and immunocytochemical properties. American Journal of Physiology, 1994. 266(6): p. C1795-C1802. 5. Watson, T., Therapeutic Ultrasound. 2017. 6. Gail ter Haar, Review Therapeutic ultrasound, European Journal of Ultrasound Volume 9, Issue 1, 1999, Pages 3-9. 7. Kyoko Ikeda, Tadahiro Takayama, Naoto Suzuki, Koichi Shimada, Kichibee Otsuka, Koichi Ito, Effects of low-intensity pulsed ultrasound on the differentiation of C2C12 cells, Life Sciences, Volume 79, Issue 20, 2006, Pages 1936-1943, 8. Puts, R., et al., Activation of Mechanosensitive Transcription Factors in Murine C2C12 Mesenchymal Precursors by Focused Low-Intensity Pulsed Ultrasound (FLIPUS). Ieee Transactions on Ultrasonics Ferroelectrics and Frequency Control, 2016. 63(10): p. 1505-1513 9. Salgarella, A.R., et al., Optimal ultrasound exposure conditions for maximizing c2c12 muscle cell proliferation and differentiation. Ultrasound in Medicine and Biology, 2017. 43(7): p. 1452-1465. 10. Fung, C.H., et al., Osteocytes exposed to far field of therapeutic ultrasound promotes osteogenic cellular activities in pre-osteoblasts through soluble factors. Ultrasonics, 2014. 54(5): p. 1358-1365. 11. Imashiro, C., et al., Cell Patterning Method on a Clinically Ubiquitous Culture Dish Using Acoustic Pressure Generated From Resonance Vibration of a Disk-Shaped Ultrasonic Transducer. Ieee Transactions on Biomedical Engineering, 2019. 66(1): p. 111-118 12. O'Brien, W.D., Ultrasound-biophysics mechanisms. Progress in Biophysics Molecular Biology, 2007. 93(1-3): p. 212-255 13. Han Y, Liu C, Zhang D, Men H, Huo L, Geng Q, Wang S, Gao Y, Zhang W, Zhang Y, Zhang Y, et al: Mechanosensitive ion channel Piezo1 promotes prostate cancer development through the activation of the Akt/mTOR pathway and acceleration of cell cycle. Int J Oncol 55: 629-644, 2019 14. Secomski, W., et al., In vitro ultrasound experiments: Standing wave and multiple reflections influence on the outcome. Ultrasonics, 2017. 77: p. 203-213. 15. Sushma, Amal Chandra Mondal, Role of GPCR signaling and calcium dysregulation in Alzheimer's disease, Molecular and Cellular Neuroscience, Volume 101, 2019, 103414, 16. ZHANG, W., LIU, H. MAPK signal pathways in the regulation of cell proliferation in mammalian cells. Cell Res 12, 9–18 (2002). 17. Agell N, Bachs O, Rocamora N, Villalonga P. Modulation of the Ras/Raf/MEK/ERK pathway by Ca(2+), and calmodulin. Cell Signal. 2002 Aug;14(8):649-54. 18. Tsuchiya, M., Hara, Y., Okuda, M. et al. Cell surface flip-flop of phosphatidylserine is critical for PIEZO1-mediated myotube formation. Nat Commun 9, 2049 (2018). 19. Liao, D., Hsiao, MY., Xiang, G. et al. Optimal pulse length of insonification for Piezo1 activation and intracellular calcium response. Sci Rep 11, 709 (2021). 20. Hensel, K., M.P. Mienkina, and G. Schmitz, ANALYSIS OF ULTRASOUND FIELDS IN CELL CULTURE WELLS FOR IN VITRO ULTRASOUND THERAPY EXPERIMENTS. Ultrasound in Medicine and Biology, 2011. 37(12): p. 2105-2115. 21. Fung, C.H., et al., Investigation of rat bone fracture healing using pulsed 1.5 MHz, 30 mW/cm(2) burst ultrasound - Axial distance dependency. Ultrasonics, 2014. 54(3): p. 850-859. 22. Sun, C., Yuan, H., Wang, L., Wei, X., Williams, L., Krebsbach, P.H., Guan, J.-L. and Liu, F. (2016), FAK Promotes Osteoblast Progenitor Cell Proliferation and Differentiation by Enhancing Wnt Signaling. J Bone Miner Res, 31: 2227-2238. 23. Ni, Y, Wang, X, Yin, X, et al. Plectin protects podocytes from adriamycin-induced apoptosis and F-actin cytoskeletal disruption through the integrin α6β4/FAK/p38 MAPK pathway. J Cell Mol Med. 2018; 22: 5450– 5467. 24. Ju Guang Wang, Motoi Miyazu, Peng Xiang, Shu Nong Li, Masahiro Sokabe, Keiji Naruse, Stretch-induced cell proliferation is mediated by FAK-MAPK pathway, Life Sciences, Volume 76, Issue 24, 2005, Pages 2817-2825. 25. Saleem, S., Li, J., Yee, S.-P., Fellows, G.F., Goodyer, C.G. and Wang, R. (2009), β1 integrin/FAK/ERK signalling pathway is essential for human fetal islet cell differentiation and survival. J. Pathol., 219: 182-192. 26. Conejo, R. and Lorenzo, M. (2001), Insulin signaling leading to proliferation, survival, and membrane ruffling in C2C12 myoblasts. J. Cell. Physiol., 187: 96-108. 27. Ju-Hye Lee, Hirofumi Tachibana, Yoshiko Morinaga, Yoshinori Fujimura, Koji Yamada,Modulation of proliferation and differentiation of C2C12 skeletal muscle cells by fatty acids, Life Sciences, Volume 84, Issues 13–14, 2009, Pages 415-420. 28. Buel D. Rodgers, Benjamin D. Wiedeback, Knut E. Hoversten, Melissa F. Jackson, Ryan G. Walker, Thomas B. Thompson, Myostatin Stimulates, Not Inihibits, C2C12 Myoblast Proliferation, Endocrinology, Volume 155, Issue 3, 1 March 2014, Pages 670–675. 29. Abmayr SM, Pavlath GK. Myoblast fusion: lessons from flies and mice. Development. 2012 Feb;139(4):641-56. 30. Engler, A.J., et al., Myotubes differentiate optimally on substrates with tissue-like stiffness: pathological implications for soft or stiff microenvironments. Journal of Cell Biology, 2004. 166(6): p. 877-887. 31. Zheng J, et al. Pterostilbene Enhances Endurance Capacity via Promoting Skeletal Muscle Adaptations to Exercise Training in Rats. Molecules 25:N/A (2020). 32. Mañas-García L et al. Muscle Phenotype, Proteolysis, and Atrophy Signaling During Reloading in Mice: Effects of Curcumin on the Gastrocnemius. Nutrients 12:N/A (2020). 33. Xu, T., Su, J., Jiang, X., Rehrig, P., Hackenberger, W. (2005). The Load Capability of Piezoelectric Single Crystal Actuators. MRS Proceedings, 888, 0888-V08-02. doi:10.1557/PROC-0888-V08-02 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/80863 | - |
| dc.description.abstract | 近20年研究肌肉再生之研究多不勝數，無論是電刺激還是化學藥劑刺激以至超音波刺激，眾多研究皆發現不同的刺激方式能激發不一樣的內在調控，特別是肌肉再生，其中低能量脈衝超音波(LIPUS)引導C2C12肌母細胞之增生與分化皆有促進的效果。本實驗以C2C12肌母細胞做為體外培養之骨骼肌模型，透過比較不同能量之超音波以及時間，挑選最佳的參數對C2C12肌母細胞施打後能使其快速分化成肌管。 經由增生實驗比較出在每日刺激5分鐘、施打能量為23.12 mW/cm2後，C2C12肌母細胞的增生數量相對於控制組能有顯著差異，以此能量做西方墨點轉漬法分析也可得到超音波刺激的組別p-ERK含量也高於控制組。 為了瞭解其原因，我們嘗試透過鈣離子實驗(Calcium)去理解肌母及肌管不同的內在調控機制。透過3種不同的低能量脈衝超音波刺激參數(高聲壓與高聲流之混合條件、高聲流為主的條件以及高聲壓的條件)與抑制離子通道及GPCR後可得知肌管的鈣離子調控反應會因不同的條件以及對應之抑制劑而被抑制；肌母細胞則否，這表明在分化之過程中肌母的一些機械式受器(Mechanoreceptors)並無繼續被保留，某些機械式受器(Mechanoreceptors)也因功能性分化而有增多的現象。 最後根據鈣離子實驗之結果，對其下Src抑制劑去影響FAK等活性，對其施打比增生實驗更強之低能量脈衝超音波能量(61.52 mW/cm2)後做西方墨點轉漬法分析發現當其被抑制後p-ERK也會降低而未抑制的組別則會提高。使用微圖案(Micropattern)去限制C2C12肌母細胞生長的區域，固定其分化成肌管後的方向一致性，施打同樣的超音波能量後，對其做螢光染色分析分化過程中之肌管長度與核融合比例(Fusion index)後，發現兩者相較於控制組、超音波組皆有顯著的差異(p<0.001)。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T03:19:33Z (GMT). No. of bitstreams: 1 U0001-2709202116064800.pdf: 4061140 bytes, checksum: 852ca9899895d478b88c6e4e9c597041 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | 第一章、緒論 - 1 - 1.1 肌少症(Sarcopenia) - 1 - 1.2 肌肉生成機制 - 1 - 1.2.1 衛星細胞(Satellite cells) - 1 - 1.2.2 C2C12體外肌肉模型 - 1 - 1.3 醫用超音波 - 2 - 1.4 超音波之定義 - 3 - 1.4.1 超音波之脈衝重複頻率(PRF)、涵蓋率(Duty cycle) - 3 - 1.4.2 超音波之空間能量分布 - 3 - 1.5 LIPUS刺激C2C12之研究 - 4 - 1.6 細胞受外來刺激之內部鈣離子訊號調控 - 4 - 1.7 Src在FAK signaling 所扮演的關鍵角色 - 6 - 第二章、材料與方法 - 7 - 2.1 超音波實驗刺激裝置 - 7 - 2.1.1 波型產生器(Function generator) - 9 - 2.1.2 功率放大器(Amplifier) - 9 - 2.1.3 超音波刺激探頭 - 10 - 2.2 實驗設備校正 - 10 - 2.2.1 沉水探頭(Hydrophone) - 10 - 2.2.2 超音波能量量測與換算方法 - 11 - 2.2.3 超音波刺激裝置升溫量測 - 12 - 2.3 C2C12肌母細胞增生與分化實驗 - 12 - 2.3.1 C2C12肌母細胞之準備 - 12 - 2.3.2 C2C12肌母細胞增生(Proliferation)實驗 - 13 - 2.3.3 C2C12肌母細胞分化(Differentiation)實驗 - 16 - 2.4 西方墨點轉漬法(Western blot) - 17 - 2.5 免疫螢光染色(Immunofluorescence) - 18 - 2.6 鈣離子調控實驗 - 20 - 2.6.1 C2C12細胞備製 - 20 - 2.6.2 鈣離子螢光染色(BAPTA-1) - 20 - 2.6.3 鈣離子調控實驗(BAPTA-1) - 20 - 2.6.4 鈣離子螢光染色(Fura-2) - 22 - 2.6.5 鈣離子調控實驗(Fura-2) - 22 - 2.6.6 鈣離子調控實驗影像分析方法 - 23 - 2.7 C2C12肌管融合實驗 - 25 - 第三章、實驗結果與討論 - 27 - 3.1 超音波裝置升溫實驗 - 27 - 3.2 C2C12 肌母細胞增生(Proliferation)實驗 - 27 - 3.2.1 C2C12 肌母細胞p-ERK實驗 - 29 - 3.3 C2C12 肌母細胞分化(Differentiation)實驗 - 30 - 3.4 鈣離子實驗(BAPTA-1) - 31 - 3.5 鈣離子實驗(Fura-2) - 32 - 3.6 C2C12肌管細胞核融合與長度實驗 - 34 - 3.7 討論 - 36 - 3.7.1 分化實驗 - 36 - 3.7.2 鈣離子實驗(BAPTA-1) - 36 - 3.7.3 鈣離子實驗(Fura-2) - 37 - 3.7.4 C2C12肌管細胞核融合與長度實驗 - 38 - 第四章、結論與未來展望 - 40 - 4.1 結論 - 40 - 4.2 未來展望 - 40 - 參考資料 - 41 - | |
| dc.language.iso | zh-TW | |
| dc.subject | 微能量超音波 | zh_TW |
| dc.subject | C2C12 肌母細胞 | zh_TW |
| dc.subject | 增生 | zh_TW |
| dc.subject | 分化 | zh_TW |
| dc.subject | 鈣離子實驗 | zh_TW |
| dc.subject | Micropattern | en |
| dc.subject | C2C12 myoblast | en |
| dc.subject | Low-intensity ultrasound | en |
| dc.subject | Proliferation | en |
| dc.subject | Differentiation | en |
| dc.subject | Calcium | en |
| dc.title | 超音波受器對肌母細胞增生與分化之影響 | zh_TW |
| dc.title | The role of mechanoreceptors in myoblast proliferation and differentiation | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 張孜菁(Hsin-Tsai Liu),陳文翔(Chih-Yang Tseng) | |
| dc.subject.keyword | C2C12 肌母細胞,微能量超音波,增生,分化,鈣離子實驗, | zh_TW |
| dc.subject.keyword | C2C12 myoblast,Low-intensity ultrasound,Proliferation,Differentiation,Calcium,Micropattern, | en |
| dc.relation.page | 43 | |
| dc.identifier.doi | 10.6342/NTU202103399 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-10-04 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
| 顯示於系所單位： | 醫學工程學研究所 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| U0001-2709202116064800.pdf 授權僅限NTU校內IP使用（校園外請利用VPN校外連線服務） | 3.97 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。