本文選題：粒毛盤菌胞外多糖結(jié)構(gòu) + 抗腫瘤活性��； 參考：《合肥工業(yè)大學(xué)》2017年碩士論文

【摘要】：本研究經(jīng)發(fā)酵提取、分離純化獲得一種新的粒毛盤菌YM130胞外多糖(LEP-2a),采用單糖組成分析、甲基化分析、傅里葉變換紅外光譜分析和核磁共振光譜分析對LEP-2a進行結(jié)構(gòu)表征,并評價其體外抗腫瘤活性;分別對LEP-2a進行乙酰化和苯甲�；揎�,并研究LEP-2a及其衍生物對STZ誘導(dǎo)的糖尿病小鼠的心血管保護作用。通過高效凝膠滲透色譜(HPGPC)檢測LEP-2a為均一組分,且平均分子量約為1.61×106 Da。單糖組成分析結(jié)果表明,LEP-2a由甘露糖和半乳糖組成,且摩爾比為3.8:1.0。甲基化分析、FT-IR分析和NMR分析結(jié)果表明,LEP-2a的主鏈由1,2-linked-α-D-Manp、1,2,6-linked-α-D-Manp、1,3,4-linked-α-D-Manp和1,3-linked-β-D-Galp組成。體外抗腫瘤活性研究表明,在濃度為100、200、300和400μg/m L時,LEP-2a能夠顯著增強5-氟尿嘧啶對Hela細(xì)胞的抑制作用。分別對LEP-2a進行乙�；捅郊柞；揎�,獲得取代度為0.220的乙�；疞EP-2a(ALEP-2a)和取代度為0.574的苯甲酰化LEP-2a(BLEP-2a)。FT-IR分析和13C NMR分析結(jié)果進一步證實了乙酰化和苯甲�；揎棾晒Α；�13C NMR分析的結(jié)果,我們推測出→2)-α-D-Manp-(1→的C-3、C-4和C-6位,α-D-Manp-(1→的C-2、C-3、C-4和C-6位,→2,6)-α-D-Manp-(1→的C-3和C-4位,→3,4)-α-D-Manp-(1→的C-2、C-4和C-6位,→3)-β-D-Galp-(1→的C-1、C-2、C-4和C-6位以及β-D-Galp-(1→的C-2、C-4和C-6位可能被乙�；糠秩〈�;→3)-β-D-Galp-(1→的C-1位被完全苯甲�；�,→2)-α-D-Manp-(1→、α-D-Manp-(1→和→2,6)-α-D-Manp-(1→的C-4位以及→2)-α-D-Manp-(1→、α-D-Manp-(1→、→3,4)-α-D-Manp-(1→、→3)-β-D-Galp-(1→和β-D-Galp-(1→的C-6位可能被苯甲�；糠秩〈Ｍㄟ^腹腔注射鏈脲佐菌素(STZ)構(gòu)建糖尿病小鼠模型,并研究不同劑量LEP-2a、ALEP-2a和BLEP-2a對糖尿病小鼠的心血管保護作用。結(jié)果表明,LEP-2a及其衍生物具有心血管保護作用且存在劑量-效應(yīng)關(guān)系;與模型組相比,治療組小鼠心臟、腎臟和肝臟指數(shù)顯著降低,心臟組織中超氧化物歧化酶(SOD)、谷胱甘肽過氧化物酶(GSH-PX)和過氧化氫酶(CAT)活性顯著增加,丙二醛(MDA)水平顯著降低,血清中晚期糖基化終產(chǎn)物(AGEs)、高敏C反應(yīng)蛋白(hs-CRP)、可溶性細(xì)胞間粘附分子1(sICAM-1)和內(nèi)皮素1(ET-1)水平顯著降低,一氧化氮(NO)水平顯著增加;與同劑量的LEP-2a相比,ALEP-2a和BLEP-2a表現(xiàn)出更顯著的心血管保護作用。此外,組織病理學(xué)觀察顯示,治療組小鼠受損心肌結(jié)構(gòu)得到改善。綜上所述,LEP-2a及其衍生物的作用機制可能與改善抗氧化防御系統(tǒng)、抑制氧化應(yīng)激以及促進受損心肌再生有關(guān)。
[Abstract]:In this study, a new extracellular polysaccharide (Lep-2aa) was isolated and purified by fermentation and purification. The structure of LEP-2a was characterized by monosaccharide composition analysis, methylation analysis, Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance spectroscopy (NMR). The antitumor activity of LEP-2a was evaluated in vitro, the acetylation and benzoylation of LEP-2a were modified, and the cardiovascular protective effects of LEP-2a and its derivatives on STZ-induced diabetic mice were studied. LEP-2a was homogenized by high performance gel permeation chromatography (HPGPC), and the average molecular weight was about 1.61 脳 106 Da. The results of monosaccharide composition analysis showed that the LEP-2a was composed of mannose and galactose, and the molar ratio was 3.8: 1.0. The results of FT-IR and NMR analysis of methylation analysis show that the main chain of LEP-2a is composed of 1k2-linked- 偽 -D-Manpan- 1, 6-linked- 偽 -D-Manpan- (34-linked- 偽 -D-Manp) and 1-3-linked- 尾 -D-Galp (1-3-linked- 尾 -D-Galp). In vitro antitumor activity studies showed that the inhibitory effect of 5-fluorouracil on Hela cells was significantly enhanced at concentrations of 100200300 渭 g / mL and 400 渭 g / mL. The results of acetylation and benzoylation of LEP-2a with a degree of substitution of 0.220 and benzoylation of LEP-2a with a degree of substitution of 0.574 were obtained respectively. The results of FT-IR and 13C NMR further confirmed that the modification of acetylation and benzoylation was successful. 鍩轟簬13C NMR鍒嗘瀽鐨勭粨鏋，

本文編號：1982594