植物甾醇氧化物及纳米核磁造影剂的细胞毒性效应研究
发布时间：2018-10-14 16:19
【摘要】：外源性化合物是人类日常生活中接触外界环境不可避免的一类化合物,这些化合物可能与机体接触并进入机体,并且在机体内产生一定的生物活性。有些化学物质可能与机体组织发生生物化学作用,破坏正常生理功能,对人类健康产生负面影响,有些化学物质也可能有益于人体健康。植物甾醇氧化物和胆固醇氧化物是存在人类日常饮食中一类化合物,大量的研究证明胆固醇氧化物对人体具有不良的生物效应,植物甾醇氧化物结构与胆固醇相似,而关于植物甾醇氧化物的生物安全性尚未定论。随着纳米技术在生物医学领域中的应用,具有多功能的纳米造影剂不断问世,造影剂作为典型的临床辅助诊断的外源性化合物,纳米造影剂的产生为造影剂的发展和应用带来便利的同时,其生物安全性研究引起了国内外许多学者的关注。目前纳米造影剂的生物安全性研究尚处于起步阶段,而且由于纳米材料种类繁多,理化性质各异,人们对于纳米造影剂毒性效应及安全评价的认识还相当局限。因此,探究这些外源性化合物对机体的生物学效应,明确其在机体内的生物安全性具有重要意义。本研究主要以两类(7-酮基和7β-羟基)植物甾醇氧化物和两种不同结构的纳米核磁造影剂为研究对象,主要研究了这些外源性化合物对不同细胞的细胞毒性效应,获得了如下主要结果:1.以胆固醇氧化物(7K-CH和7β-OH-CH)为阳性对照组,将两类植物甾醇氧化物分别作用于HIC细胞后,在120μM作用浓度下,7-酮基植物甾醇氧化物对HIC细胞的毒性强度依次为7K-SI7K-CA7K-BR7K-ST7K-MIX,7β-羟基植物甾醇氧化物对HIC细胞的毒性强度依次为7β-OH-SI7β-OH-CA7β-OH-MIX7β-OH-ST≈7β-OH-BR。HIC细胞系可能对7-酮基系列氧化物要比对7β-羟基植物甾醇氧化物更敏感。所有被测氧化物都能明显降低细胞活力,并具有浓度和时间依赖性,尤其是7K-SI和7K-CA活性最高,降低细胞活力最为明显。此外,7K-SI和7K-CA不仅诱导早期凋亡细胞比例上升,而且还导致细胞周期S期含量上升,G1期含量下降,而7K-BR、7K-ST和7K-MIX没有显示出明显的细胞毒性,与此同时,7K-SI、7K-CA和7K-BR通过激活caspase-3活性,调节Bcl-2蛋白诱导细胞凋亡,而7K-ST和7K-MIX不依赖于caspase-3和Bcl-2。2.两类甾醇氧化物作用A549细胞后,被测氧化物均降低了A549的细胞活力且具有浓度和时间依赖关系。在120μM作用浓度下,7-酮基植物甾醇氧化物对A549细胞毒性强度依次为7K-SI7K-CA7K-BR≈7K-ST≈7K-MIX,7β-羟基-植物甾醇氧化物对A549细胞的毒性强度依次为7β-OH-SI≈7β-OH-CA7β-OH-MIX7β-OH-ST≈7β-OH-BR,这一结果表明A549细胞系可能对7β-羟基系列氧化物要比对7-酮基植物甾醇氧化物更敏感,这一结果正好与HIC细胞系结果相反。另外,7β-羟基系列氧化物不仅降低了细胞活力,还导致A549凋亡后期细胞比例上升,同时细胞周期实验表明7β-羟基-CH、7β-羟基-SI、7β-羟基-CA和7K-CH导致sub-G1期的含量有所增加,但是被测氧化物对A549细胞周期的影响因化合物不同而效果不同,这也表明了甾醇氧化物对A549细胞周期与这些氧化物的化学结构没有直接的关联性。此外,这些氧化物会导致A549细胞内活性氧含量随着时间延长而上升,同时通过激活caspase-3活性,调节Bcl-2蛋白诱导细胞凋亡。3.两类甾醇氧化物作用于HepG2细胞后,被测的氧化物均降低了HepG2细胞活力并且具有浓度时间依赖关系。在120μM作用浓度下,7酮基-ST和7-酮基-MIX表现出强抑制细胞活性的特点,这一结果与HIC细胞和A549细胞实验结果有很大差异。7β-羟基-CA和7酮基-CA不仅降低细胞活性,激发超氧化物歧化酶SOD和过氧化氢酶进而导致细胞凋亡,但是这两种单体没有导致脂质过氧化物含量上升,而混合甾醇氧化物导致MDA含量略微上升,但是没有影响细胞活力。实验表明氧化应激反应与细胞死亡之间没有必然的联系,植物甾醇氧化物对HepG2细胞毒性与细胞对这些氧化物的摄取量有关系,因化合物不同而效果不同。4.本实验中采用的纳米材料Fe_3O_4@HSiO_2和负载了秋水仙碱的Fe_3O_4@HSiO_2-COLC是具抗肿瘤药物载体和核磁造影剂功能的磁性纳米材料体系。核磁共振成像实验表明,纳米材料Fe_3O_4@HSiO_2作为核磁造影剂没有显示出细胞毒性,同时具有很好的生物相容性和较高的核磁对比度,作为抗癌药物载体(Fe_3O_4@HSiO_2 COLC)能在酸性环境中(pH3.0)有效释放所负载的秋水仙碱(COLC),在磁场作用下能有效地靶向肿瘤细胞HepG2而不损伤正常细胞,且药效要高于单纯的COLC。5.由聚乙烯亚胺包裹叶酸和钆介孔2-50 nm的纳米材料体系Gd-FA-Si,具有较好的生物相容性,兼具抗肿瘤药物载体和核磁造影剂功能,同时受pH调控而释放药物。纳米材料表面负载的叶酸能高效的靶向Hela和MDA-MB-231细胞上的叶酸受体,材料负载的阿霉素高效抑制细胞活性,比单纯的阿霉素药物给药具有更高的药效。
[Abstract]:Exogeneous compound is a kind of compound that comes into contact with the outside world in daily life. These compounds may come into contact with the organism and enter the organism, and produce certain biological activity in the organism. Some chemicals may have biochemical effects with body tissues, disrupt normal physiological functions, have negative effects on human health, and some chemicals may also be beneficial to human health. Phytochrome oxide and cholesterol oxide are a type of compound in the daily diet of humans, and a large number of studies have shown that cholesterol oxides have poor biological effects on the human body, and that the plant's oxide structure is similar to that of cholesterol. However, the biological safety of phytochrome oxide has not been decided yet. With the application of nanotechnology in the field of biomedicine, the multifunctional nano contrast agent is constantly invented, the contrast agent is taken as an exogenous compound for the typical clinical assistant diagnosis, and the production of the nano contrast agent brings convenience to the development and application of the contrast agent, Its biological safety research has attracted the attention of many scholars both at home and abroad. At present, the biological safety research of nano-contrast agent is still in the initial stage, and because of the wide variety of nano-materials and different physical and chemical properties, people's understanding of the toxicity effect and safety evaluation of the nano-contrast agent is quite limited. Therefore, it is important to explore the biological effect of these exogenous compounds on organism and to clarify its biological safety in organism. In this study, two kinds of nano-nuclear magnetic contrast agents (7-keto-and 7-hydroxy group) and two kinds of nano-nuclear magnetic contrast agents were studied. The cytotoxicity effect of these exogenous compounds on different cells was studied, and the following main results were obtained: 1. Using cholesterol oxide (7K-CH and 7H2O-OH-CH) as the positive control group, the toxicity intensity of the 7-keto-base plant oxide was 7K-SI7K-CA7K-BR7K-ST7K-MIX, respectively. The toxicity intensity of 7-keto-OH-SI7 CHA-OH-CA7 CHA-OH-MICACHE-OH-ST-7OH-OH-BR. 7721 cell line may be more sensitive to 7-keto-series oxides than for 7-keto-hydroxyphytochrome oxide. All the tested oxides can obviously decrease the activity of cells, and have the highest concentration and time dependence, especially the highest activity of 7K-SI and 7K-CA and the most obvious decrease of cell viability. In addition, 7K-SI and 7K-CA not only induce an increase in the proportion of early apoptotic cells but also lead to an increase in the S phase of the cell cycle and a decrease in G1 phase, while 7K-BR, 7K-ST and 7K-MIX show no significant cytotoxicity, while 7K-SI, 7K-CA and 7K-BR activate caspase-3 activity, Bcl-2 protein was regulated to induce apoptosis, while 7K-ST and 7K-MIX were independent of caspase-3 and Bcl-2. After A549 cells were treated with two kinds of exogenous oxides, the measured oxides reduced the viability of A549 cells and had a concentration and time-dependent relationship. The toxic strength of the 7-keto-phytochrome oxide on A549 cells was 7K-SI7K-CA7K-BR-7K-ST-7K-MIX in order of 120. m This result shows that the A549 cell line may be more sensitive to the 7-keto-group oxide than to the 7-keto-based plant, and this result is just contrary to the results of the cell line. in addition, that 7-hydroxy-hydroxy-series oxide not only reduce the activity of the cell, but also lead to the increase of the cell proportion in the late stage of apoptosis of the A549 cell, and meanwhile, the cell cycle experiment shows that the content of the sub-G1 phase is increased due to the 7-hydroxy-CH, 7OH-hydroxy-SI, 7OH-hydroxy-CA and 7K-CH, However, the effect of oxide on the cell cycle of A549 is different because of the different compounds, which also indicates that the cell cycle of the A549 cells is not directly related to the chemical structure of these oxides. In addition, these oxides lead to increased active oxygen content in A549 cells over time, while modulating Bcl-2 protein-induced apoptosis by activating caspase-3 activity. Both of the two types of oxygenic oxide have reduced the activity of HepG2 cells and have a concentration-time dependent relationship after HepG2 cells. At the concentration of 120. m u.M, the 7-keto-ST and 7-keto-MIX exhibit strong inhibitory cellular activity. The result is very different from the experimental results of Jurkat cells and A549 cells. The 7-keto-hydroxy-CA and 7-keto-CA not only decrease cell activity, Activation of superoxide dismutase (SOD) and catalase in turn led to apoptosis, but the two monomers did not cause the lipid peroxide content to rise, but the mixed oxide resulted in a slight increase in MDA content, but did not affect the viability of the cells. The results showed that there was no definite relationship between oxidative stress response and cell death. The toxicity of phytochrome oxide to HepG2 cells was related to the uptake of these oxides. Fe _ 3O _ 4 @ HSiO _ 2 and Fe _ 3O _ 4 @ HSiO _ 2-COLC, which are used in this experiment, are magnetic nano-materials with anti-tumor drug carrier and nuclear magnetic contrast agent. The nuclear magnetic resonance imaging experiments show that the nano-material Fe _ 3O _ 4 @ HSiO _ 2 has not shown cytotoxicity as a nuclear magnetic contrast agent, but also has good biocompatibility and high nuclear magnetic contrast. 浣滀负鎶楃檶鑽墿杞戒綋(Fe_3O_4@HSiO_2 COLC)鑳藉湪閰告，
本文编号：2270977
[Abstract]:Exogeneous compound is a kind of compound that comes into contact with the outside world in daily life. These compounds may come into contact with the organism and enter the organism, and produce certain biological activity in the organism. Some chemicals may have biochemical effects with body tissues, disrupt normal physiological functions, have negative effects on human health, and some chemicals may also be beneficial to human health. Phytochrome oxide and cholesterol oxide are a type of compound in the daily diet of humans, and a large number of studies have shown that cholesterol oxides have poor biological effects on the human body, and that the plant's oxide structure is similar to that of cholesterol. However, the biological safety of phytochrome oxide has not been decided yet. With the application of nanotechnology in the field of biomedicine, the multifunctional nano contrast agent is constantly invented, the contrast agent is taken as an exogenous compound for the typical clinical assistant diagnosis, and the production of the nano contrast agent brings convenience to the development and application of the contrast agent, Its biological safety research has attracted the attention of many scholars both at home and abroad. At present, the biological safety research of nano-contrast agent is still in the initial stage, and because of the wide variety of nano-materials and different physical and chemical properties, people's understanding of the toxicity effect and safety evaluation of the nano-contrast agent is quite limited. Therefore, it is important to explore the biological effect of these exogenous compounds on organism and to clarify its biological safety in organism. In this study, two kinds of nano-nuclear magnetic contrast agents (7-keto-and 7-hydroxy group) and two kinds of nano-nuclear magnetic contrast agents were studied. The cytotoxicity effect of these exogenous compounds on different cells was studied, and the following main results were obtained: 1. Using cholesterol oxide (7K-CH and 7H2O-OH-CH) as the positive control group, the toxicity intensity of the 7-keto-base plant oxide was 7K-SI7K-CA7K-BR7K-ST7K-MIX, respectively. The toxicity intensity of 7-keto-OH-SI7 CHA-OH-CA7 CHA-OH-MICACHE-OH-ST-7OH-OH-BR. 7721 cell line may be more sensitive to 7-keto-series oxides than for 7-keto-hydroxyphytochrome oxide. All the tested oxides can obviously decrease the activity of cells, and have the highest concentration and time dependence, especially the highest activity of 7K-SI and 7K-CA and the most obvious decrease of cell viability. In addition, 7K-SI and 7K-CA not only induce an increase in the proportion of early apoptotic cells but also lead to an increase in the S phase of the cell cycle and a decrease in G1 phase, while 7K-BR, 7K-ST and 7K-MIX show no significant cytotoxicity, while 7K-SI, 7K-CA and 7K-BR activate caspase-3 activity, Bcl-2 protein was regulated to induce apoptosis, while 7K-ST and 7K-MIX were independent of caspase-3 and Bcl-2. After A549 cells were treated with two kinds of exogenous oxides, the measured oxides reduced the viability of A549 cells and had a concentration and time-dependent relationship. The toxic strength of the 7-keto-phytochrome oxide on A549 cells was 7K-SI7K-CA7K-BR-7K-ST-7K-MIX in order of 120. m This result shows that the A549 cell line may be more sensitive to the 7-keto-group oxide than to the 7-keto-based plant, and this result is just contrary to the results of the cell line. in addition, that 7-hydroxy-hydroxy-series oxide not only reduce the activity of the cell, but also lead to the increase of the cell proportion in the late stage of apoptosis of the A549 cell, and meanwhile, the cell cycle experiment shows that the content of the sub-G1 phase is increased due to the 7-hydroxy-CH, 7OH-hydroxy-SI, 7OH-hydroxy-CA and 7K-CH, However, the effect of oxide on the cell cycle of A549 is different because of the different compounds, which also indicates that the cell cycle of the A549 cells is not directly related to the chemical structure of these oxides. In addition, these oxides lead to increased active oxygen content in A549 cells over time, while modulating Bcl-2 protein-induced apoptosis by activating caspase-3 activity. Both of the two types of oxygenic oxide have reduced the activity of HepG2 cells and have a concentration-time dependent relationship after HepG2 cells. At the concentration of 120. m u.M, the 7-keto-ST and 7-keto-MIX exhibit strong inhibitory cellular activity. The result is very different from the experimental results of Jurkat cells and A549 cells. The 7-keto-hydroxy-CA and 7-keto-CA not only decrease cell activity, Activation of superoxide dismutase (SOD) and catalase in turn led to apoptosis, but the two monomers did not cause the lipid peroxide content to rise, but the mixed oxide resulted in a slight increase in MDA content, but did not affect the viability of the cells. The results showed that there was no definite relationship between oxidative stress response and cell death. The toxicity of phytochrome oxide to HepG2 cells was related to the uptake of these oxides. Fe _ 3O _ 4 @ HSiO _ 2 and Fe _ 3O _ 4 @ HSiO _ 2-COLC, which are used in this experiment, are magnetic nano-materials with anti-tumor drug carrier and nuclear magnetic contrast agent. The nuclear magnetic resonance imaging experiments show that the nano-material Fe _ 3O _ 4 @ HSiO _ 2 has not shown cytotoxicity as a nuclear magnetic contrast agent, but also has good biocompatibility and high nuclear magnetic contrast. 浣滀负鎶楃檶鑽墿杞戒綋(Fe_3O_4@HSiO_2 COLC)鑳藉湪閰告，
本文编号：2270977
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