本文選題：發(fā)酵樺褐孔菌 + HepG-2細(xì)胞��； 參考：《延邊大學(xué)》2017年碩士論文

【摘要】：目的初步探討發(fā)酵樺褐孔菌對(duì)肝癌HepG-2細(xì)胞的凋亡作用。方法分別采用硫酸苯酚法、Folin-Ciocalteu法、比色法測定未發(fā)酵、發(fā)酵樺褐孔多糖、萜類和多酚的含量;以肝癌HepG-2細(xì)胞為實(shí)驗(yàn)對(duì)象,采用MTT法測定發(fā)酵樺褐孔菌對(duì)肝癌HepG-2細(xì)胞增殖的影響;HE染色法觀察發(fā)酵樺褐孔菌對(duì)肝癌HepG-2細(xì)胞形態(tài)學(xué)的影響;流式細(xì)胞儀測定發(fā)酵樺褐孔菌對(duì)肝癌HepG-2細(xì)胞凋亡作用及細(xì)胞周期的影響;瓊脂糖凝膠電泳法觀察發(fā)酵樺褐孔菌對(duì)肝癌HepG-2細(xì)胞DNA裂解的影響。結(jié)果與未發(fā)酵樺褐孔菌比較,發(fā)酵樺褐孔菌多糖、多酚和萜類含量明顯升高(P0.01);發(fā)酵樺褐孔菌顯著抑制肝癌HepG-2細(xì)胞增殖,其抑制作用與給藥時(shí)間和給藥濃度呈正相關(guān),且與未發(fā)酵樺褐孔菌相比,發(fā)酵樺褐孔菌濃度為100mg/L,處理時(shí)間為48h對(duì)肝癌細(xì)胞的抑制作用更明顯(P0.05),且呈時(shí)間與劑量依賴性;與對(duì)照組比較,發(fā)酵樺褐孔菌組肝癌HepG-2細(xì)胞細(xì)胞間距增大、細(xì)胞核固縮、染色加深、細(xì)胞膜皺縮、形成凋亡小體,出現(xiàn)典型凋亡形態(tài),凋亡程度隨著劑量和時(shí)間的增加而增強(qiáng),與未發(fā)酵組相比,肉眼觀察無差異;發(fā)酵樺褐孔菌可顯著誘導(dǎo)肝癌HepG-2細(xì)胞凋亡,并且隨著濃度的增加其誘導(dǎo)凋亡作用增強(qiáng),與未發(fā)酵樺褐孔菌相比,當(dāng)發(fā)酵樺褐孔菌濃度為200mg/L、400mg/L,誘導(dǎo)肝癌細(xì)胞凋亡作用更顯著(P0.01);與對(duì)照組相比,發(fā)酵樺褐孔菌組肝癌HepG-2細(xì)胞細(xì)胞周期分布受到影響,G0/G1期細(xì)胞比率顯著升高(P0.05),S期和G2/M期比率顯著下降(P0.05),與未發(fā)酵樺褐孔菌組相比,發(fā)酵樺褐孔菌濃度為200mg/L,作用時(shí)間為48h時(shí)G0/G1期細(xì)胞周期分布存在顯著性差異(P0.05);與對(duì)照組相比,發(fā)酵樺褐孔菌作用48h肝癌HepG-2細(xì)胞DNA均發(fā)生裂解,呈現(xiàn)典型的梯形凋亡形態(tài),與未發(fā)酵樺褐孔菌相比,發(fā)酵樺褐孔菌DNA裂解更明顯。結(jié)論發(fā)酵樺褐孔菌可顯著誘導(dǎo)肝癌HepG-2細(xì)胞凋亡,且與未發(fā)酵樺褐孔菌相比,發(fā)酵樺褐孔菌更顯著誘導(dǎo)肝癌HepG-2細(xì)胞凋亡。
[Abstract]:Objective to investigate the apoptosis of HepG-2 cells by fermenting Betula obliquus. Methods Folin-Ciocalteu method and colorimetric method were used to determine the contents of unfermented and fermented birch brownhole polysaccharides, terpenoids and polyphenols respectively. The effect of Betula obliquus fermentation on the proliferation of HepG-2 cells was determined by MTT method. The morphological changes of HepG-2 cells were observed by HE staining. The apoptosis and cell cycle of HepG-2 cells were determined by flow cytometry, and the DNA cleavage of HepG-2 cells was observed by agarose gel electrophoresis. Results compared with unfermented Betula obliquus, the contents of polysaccharides, polyphenols and terpenes in fermented Betula obliquus increased significantly (P 0.01), and the inhibition of P0.01C was positively correlated with the time of administration and the concentration of the drug. Compared with unfermented Betula obliquus, the concentration of 100 mg / L and the treatment time of 48 h had more obvious inhibitory effect on hepatoma cells in a time and dose-dependent manner, compared with the control group. In the fermentative group, the HepG-2 cell spacing increased, the cell nuclear pyknosis and staining deepened, the cell membrane shrank and formed apoptotic bodies, and the apoptotic degree increased with the increase of dose and time. Compared with the unfermented group, there was no significant difference between the unfermented group and the unfermented group, and the apoptosis of the HepG-2 cells was significantly induced by the fermented Betula obliquus, and the apoptotic effect was enhanced with the increase of the concentration, compared with that of the unfermented Betula obliquus. When the concentration of fermenting Betula obliquus was 200 mg / L, 400 mg / L, the apoptotic effect of hepatoma cells was more significant than that of control group. The cell cycle distribution of HepG-2 cells in fermenting Betula obliquus group was significantly increased in G _ 0 / G _ 1 phase, the ratio of P0.05 / M phase and G _ 2 / M phase was significantly decreased compared with that of unfermented Betula platyphylla group. There was a significant difference in cell cycle distribution of HepG-2 cells in G0/G1 phase at the concentration of 200mg 路L ~ (-1) and the time of treatment for 48 h. Compared with the control group, the DNA of hepatoma HepG-2 cells were lysed at 48 h after treatment by the fermenting strains of Betula obliquus, which showed typical ladder apoptotic morphology. The DNA cleavage of fermented Betula obliquus was more obvious than that of unfermented Betula obliquus. Conclusion the apoptosis of hepatoma HepG-2 cells was significantly induced by fermenting Betula obliquus, and the apoptosis of HepG-2 cells was more obvious than that of unfermented Betula obliquus.
【學(xué)位授予單位】：延邊大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R285

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