天然植物抗菌液对宫颈癌Hela细胞的杀伤作用及初步机制研究

发布时间：2018-06-13 09:13

本文选题：天然植物抗菌液 + PAMs　；参考：《西南交通大学》2017年硕士论文

【摘要】：宫颈癌是全球范围内威胁妇女生命健康的一大问题。宫颈癌的前期因其无症状而难以检测,致死率居女性恶性肿瘤第四位。宫颈癌的发病常常伴随着高危型人乳头瘤病毒(HPV)的感染,HPV极易感染人表皮和粘膜细胞,低危型可引起生殖疣类皮肤疾病,高危型HPV则可引起宫颈癌,其中高危型HPV以16和18亚型最为常见。民族医药复方制剂因其较小的毒副作用而收到广泛关注,具有良好的开发前景和研究意义。首先,采用生物信息学手段,对HPV18亚型的E6蛋白进行同源建模、对HPV18 E6和HPV16E6进行了分子动力学模拟,发现关键LOOP环起到"门控"作用:控制小分子或多肽配体进出蛋白结合位点,为药物治疗HPV感染的宫颈癌提供了理论依据。考虑天然植物抗菌液是由红花、紫草、艾纳香、刺天茄等药用植物制成的乙醇提取液。其成分复杂,作用广谱,但是具体功效成分尚不清楚。本文根据天然植物抗菌液在抗肿瘤、皮肤修复的作用以及药材来源,利用高效液相色谱技术,对轻基红花黄色素A和乌头碱进行了含量测定,测定含量分别为33.87± 1.01μg/ml和22.06土0.06μg/ml,在样品中未能检测到草屋甲素、紫草素、龙血素A、薯蓣皂苷元。其次,以M13噬菌体为病毒模型,测定天然植物抗菌液K3对M13噬菌体的抑制作用,结果表明,天然植物抗菌液K3对噬菌体有强烈的抑制效果,且在5%浓度下,抑制率达到72.38%。以Hela细胞为研究对象,初步研究了天然植物抗菌液对肿瘤细胞的抑制效果。形态学观察和CCK-8法发现天然植物抗菌液对肿瘤细胞有较强的抑制增殖和杀伤作用,与空白组相比,统计学上有极显著差异,在5%浓度下,48 h抑制率可达74.25%;划痕实验和Transwell小室迁移实验结果表明,天然植物抗菌液对肿瘤细胞体外迁移实验有较好的抑制作用,且在统计学上有显著性差异,抑制率可达65.9%。最后,通过构建裸鼠宫颈癌Hela细胞移植瘤模型,对天然植物抗菌液体内作用进行研究。发现天然植物抗菌液能够减缓肿瘤生长速度,脏器指数结果显示,其较模型组没有显著性差异;更且,天然植物抗菌液能够下调血清中的VEGF,抑制肿瘤生长。其进一步的分子作用机理尚待深入研究。
[Abstract]:Cervical cancer is a global threat to women's lives and health. The precancerous stage of cervical cancer is difficult to detect because of its asymptomatic, and the mortality is the fourth in female malignant tumor. The incidence of cervical cancer is often accompanied by high risk human papillomavirus (HPV) infection. HPV can easily infect human epidermis and mucosal cells. Low risk type can cause genital warts skin disease, high risk HPV can cause cervical cancer. High-risk HPV subtypes 16 and 18 are the most common. Because of its small toxicity and side effects, ethnic medicine compound preparation has received wide attention, and has a good development prospect and research significance. Firstly, the E6 protein of HPV18 subtype was modeled by bioinformatics, and the molecular dynamics of HPV18 E6 and HPV16 E6 were simulated. It was found that the key loop of loop acts as "gating": controlling the binding sites of small molecules or peptide ligands, which provides a theoretical basis for drug therapy of cervical cancer infected with HPV. Natural plant antibacterial solution is an ethanol extract from medicinal plants such as safflower, porphyra yezoensis, Ainarum, and Solanum. Its composition is complex, the action is broad, but the specific effect component is not clear. Based on the effect of natural plant antibacterial solution on anti-tumor, skin repair and source of medicinal materials, the contents of light safflower yellow pigment A and aconitine were determined by HPLC. The contents were 33.87 卤1.01 渭 g/ml and 22.06 卤0.06 渭 g / ml, respectively. Secondly, the bacteriophage M13 was used as a virus model to determine the inhibitory effect of natural plant antibacterial solution K3 on M13 phage. The results showed that natural plant antibacterial solution K3 had strong inhibitory effect on bacteriophage, and the inhibition rate reached 72.38% at 5% concentration. The inhibitory effect of natural plant antibacterial solution on tumor cells was studied. Morphological observation and CCK-8 method showed that the natural plant antibacterial solution could inhibit the proliferation and kill of tumor cells, and there was a significant difference between the control group and the control group. The results of scratch test and Transwell chamber migration test showed that the natural plant antibacterial solution had a good inhibitory effect on tumor cell migration in vitro, and there was significant difference in statistics, and the inhibition rate could reach 65.9%. Finally, the effect of natural plant antibacterial fluid was studied by constructing Hela cell transplantation tumor model in nude mice. The results of visceral index showed that there was no significant difference compared with the model group, moreover, the natural plant antibacterial liquid could down-regulate the VEGF in serum and inhibit tumor growth. Its further molecular mechanism needs to be further studied.
【学位授予单位】：西南交通大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.33

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