云南不同地区年青褐煤黄腐酸钠对血管新生及癌细胞增殖的影响

发布时间：2018-05-05 16:51

本文选题：云南褐煤 + 黄腐酸钠　；参考：《昆明理工大学》2014年硕士论文

【摘要】：黄腐酸是一类分子结构复杂的有机弱酸混合物,其来源不同,腐化过程不同,提取工艺不同等均有可能造成其化学和药理学活性上的差异,这极大的限制了其在药用上的研究进展,因此很有必要在对黄腐酸的来源、提纯、标准化和治疗机制上下功夫。传统用药研究认为黄腐酸具有抗溃疡、改善微循环、止血活血等功能,其机制可能与促血管新生有关,褐煤黄腐酸对血管新生的影响实际如何还未见报道,此外黄腐酸对癌细胞增殖影响的报道具有一定的争议性,有研究认为黄腐酸能促进癌细胞的增殖和转移,也有研究认为黄腐酸能抑癌抗氧化,这是否与其来源及血管新生相关也有待探讨。以此为依据本文对云南资源丰富的年青褐煤黄腐酸在同等条件下按照地域的不同采取了多样本研究,探讨了各地黄腐酸钠对血管新生的影响,以及黄腐酸钠的体外抗癌活性,经研究比较得出各地黄腐酸钠药理活性的特性,为云南各地褐煤资源能依据实际情况采取针对性的开发利用提供一定的基础数据。本论文的研究以云南峨山、寻甸、昭通三地的年青褐煤为原料,采用H202氧化降解法制备,001×7型阳离子交换树脂纯化,得到终产物精制黄腐酸。利用体外人脐静脉内皮细胞(Human umbilical vein endothelial cells, HUVEC) MTT、划痕、血管内皮细胞生长因子(Vascular edothelial growth factor, VEGF)表达测定实验,在体鸡胚绒毛尿囊膜(Chick chorioallantoic membrane, CAM)模型血管生成实验,测定各地黄腐酸钠对血管新生的影响及可能机制。此外本文还采用了MTT法测定各地黄腐酸钠对人非小细胞肺癌细胞株A549和人食管癌细胞株KYSE510的影响。主要研究结果如下： 1.峨山和寻甸的黄腐酸钠盐在HUVEC模型上的趋势具有一定的相似性,在低浓度时对HUVEC的增殖和迁移具有促进作用,同时能上调VEGF的表达,随着浓度的增高HUVEC的增殖和迁移受到抑制,分泌的VEGF浓度也降低。但两者同时也存在一定的差异性,峨山黄腐酸钠在50μg·ml-1时出现的促进效应最强,寻甸在25μg·ml-1使出现的促进效应最强,同时无论是促进还是抑制寻甸黄腐酸钠的作用强于峨山黄腐酸钠。昭通地区的褐煤黄腐酸钠在低浓度时对HUVEC没有明显的作用,在高浓度时有抑制效应,该作用与峨山地区的褐煤黄腐酸钠相当。 2.CAM实验统计结果显示云南峨山地区的褐煤黄腐酸钠在200μg·ml-1、400μg·ml-1时对CAM血管新生具有一定的促进作用,当浓度达到1200μg·ml-1时能够显著抑制CAM血管新生。寻甸褐煤黄腐酸钠在200μg·ml-1时能明显促进CAM血管新生,当浓度达到800μg·ml;-1时能显著抑制CAM血管新生。昭通地区的褐煤黄腐酸钠盐不具备促进CAM血管新生的作用,当浓度达到800μg·ml-1时可抑制CAM血管新生。三者比较无论促进作用还是抑制作用以寻甸黄腐酸钠的效应最强。 3.人非小细胞肺癌细胞A549MTT结果显示,峨山、寻甸的褐煤黄腐酸钠对A549的增殖具有一定的抑制作用,且随着浓度的增大时间的延长药物效应越强,两者比较峨山黄腐酸钠的活性强于寻甸,昭通地区的黄腐酸钠对A549几乎无效应,当浓度增大到800μg.ml-1时间延长至42h或72h时才呈现一定的抑制作用。人食管癌细胞KYSE510MTT实验结果显示,寻甸褐煤黄腐酸钠在低浓度50μg·ml-1、100μg·ml-1时反而能促进细胞的增殖,随着浓度的增大到400μg.m1-1时才出现抑制效应,峨山、昭通两地的褐煤黄腐酸钠能抑制细胞的增长,随着浓度的增大时间的延长药物效应越强。上述研究结果表明,三地的褐煤黄腐酸钠对血管新生的影响不尽相同,对实验所选取的两株癌细胞的作用也呈现出一定的差异性,且黄腐酸钠促进血管新生与促进癌细胞增殖之间并没有必然的联系,本研究进一步的说明了黄腐酸钠的药理活性研究应该以固定的产地及来源为依据,并表明了黄腐酸钠的作用机理是一个多级分、多靶点、多因素作用的过程。对于云南褐煤资源的药用开发应根据实际情况采取针对性的开采利用。
[Abstract]:Fulvic acid is a kind of complex organic weak acid mixture with different molecular structures. Its origin is different, the process of decay is different, and the different extraction processes may cause the difference in chemical and pharmacological activity, which greatly limits its research progress in medicine. Therefore, it is necessary for the source, purification, standardization and treatment machine of fulvic acid. On the basis of traditional medicine, it is believed that fulvic acid has the function of resisting ulcers, improving microcirculation, stopping blood and activating blood and so on. Its mechanism may be related to the promotion of angiogenesis, and the effect of brown coal yellow fulvic acid on angiogenesis has not yet been reported. In addition, the reports of the effect of fulvic acid on cancer cell proliferation are controversial. Fulvic acid can promote the proliferation and metastasis of cancer cells. Some studies also suggest that fulvic acid can inhibit cancer and anti-oxidation. Whether this is related to its source and angiogenesis is also to be explored.
On the basis of this, a multi sample study was taken to study the effect of sodium fulvate on angiogenesis and the anticancer activity of sodium fulvate in Yunnan under the same conditions under the same conditions, and the antitumor activity of sodium fulvate in vitro was discussed. The characteristics of the pharmacological activity of sodium fulvate in various parts of Yunnan were compared. Lignite resources can provide some basic data according to the actual situation.
The study in this paper was made of young lignite in three areas of Yunnan, Xundian and Zhaotong, prepared by H202 oxidation degradation, purified by 001 x 7 type cation exchange resin, and obtained refined fulvic acid from the final product. Using human umbilical vein endothelial cells (Human umbilical vein endothelial cells, HUVEC) MTT, scratches and vascular endothelial cell growth The Vascular edothelial growth factor (VEGF) expression test was used to determine the effect and possible mechanism of sodium fulvate on angiogenesis in the chick chorioallantoic membrane (Chick chorioallantoic membrane, CAM) model. In addition, the MTT method was used to determine the effect of sodium fulvate on human non-small cell lung cancer. The effects of cell line A549 and human esophageal cancer cell line KYSE510 were as follows:
1. the trend of the sodium fulvic acid in the 1. mount and Xundian has a certain similarity on the trend of the HUVEC model. At the low concentration, it can promote the proliferation and migration of HUVEC, and increase the expression of VEGF. With the increase of concentration, the proliferation and migration of HUVEC are inhibited, and the secretion of VEGF is also reduced. The promotion effect of Eshan sodium fulvate at 50 g. Ml-1 is the strongest. The promotion effect of Xundian in 25 mu g. Ml-1 is the strongest. At the same time, the effect of promoting or inhibiting sodium fulvate of Xundian is stronger than the sodium fulvic acid sodium. The lignite sodium fulvic acid in Zhaotong area has no obvious effect on HUVEC at low concentration, and at high concentration. The inhibition effect is similar to that of lignite in the mountain area of E Shan.
The 2.CAM experimental results show that the lignite sodium fulvate in the Yunnan area has a certain promotion effect on CAM angiogenesis at 200 g. Ml-1400 mu g. Ml-1, when the concentration reaches 1200 g. Ml-1, it can significantly inhibit the neovascularization of CAM. In the case of 200 mu g ml-1, Xundian lignite sodium fulvic can obviously promote angiogenesis, when the concentration reaches 8. 00 mu g. Ml; -1 can significantly inhibit the angiogenesis of CAM. The sodium salt of brown fulvic acid in Zhaotong area does not have the effect on promoting the angiogenesis of CAM. When the concentration reaches 800 g. Ml-1, it can inhibit the angiogenesis of CAM. The three, no matter the promotion or inhibition, the effect of sodium fulvate in Xundian is the strongest.
The results of 3. non small cell lung cancer cells A549MTT showed that the sodium fulvate of Xundian and the sodium fulvate of Xundian had a certain inhibitory effect on the proliferation of A549, and the stronger the effect of the drug effect with the time of increasing the concentration, the activity of the sodium fulvate was stronger than that in Xundian. The sodium fulvic acid in Zhaotong region had almost no effect on A549, when the concentration increased. When the time of 800 mu g.ml-1 was extended to 42h or 72h, the results of KYSE510MTT experiment in human esophageal cancer cells showed that the sodium fulvic acid of Xundian lignite could promote cell proliferation at a low concentration of 50 mu g. Ml-1100 mu g. The inhibitory effect appeared with the increase of concentration to 400 u. Lignite sodium fulvate inhibited cell growth, and the longer the concentration increased, the stronger the drug effect.
The results showed that the effect of sodium fulvate on the angiogenesis of three areas was different, and the effect on the two cancer cells selected by the experiment was also different. And the sodium fulvate promoted the angiogenesis and the proliferation of cancer cells. This study further explained the sodium fulvate. The study of pharmacological activity should be based on the fixed origin and source, and shows that the mechanism of the action of sodium fulvate is a multi-stage, multi target and multi factor process. The exploitation of Yunnan lignite resources should be exploited according to the actual situation.

【学位授予单位】：昆明理工大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96;TQ463

【参考文献】