皂角刺对肺癌的防治作用及其机制初步探讨

发布时间：2018-11-23 12:24

【摘要】：肺癌是一种对生命和人类健康威胁最大的恶性肿瘤，在我国的发病率占恶性肿瘤的首位，并逐年增加，因此对肺癌的预防和治疗作用的研究具有重要的意义。目前治疗肺癌大多采用化学药物，但在治疗过程中存在毒副作用和多药耐药性等治疗缺陷。中医药在治疗肿瘤方面有毒副作用小、能提高患者的生活质量，并且长期用药费用低等优势赢得了患者的认可，也成为开发新型抗肿瘤药物的重要源泉，因此筛选中医药抗肿瘤药物成为药理学领域研究的热点。皂角刺总黄酮是从豆科植物皂荚的干燥棘刺中提取得到的一类黄酮类化合物，各项探索结论已经证明了多种肿瘤细胞增殖能够显著地被皂角刺总黄酮所抑制。本课题探讨的目的是考察皂角刺总黄酮对肺癌细胞及肺癌模型的预防作用和治疗作用，并探讨其可能隐含的新机制。本课题体外实验采用MTT法，检测不同浓度的皂角刺总黄酮对小鼠肺癌细胞（Lewis）、小鼠胚肺成纤维细胞（L929）、小鼠脾淋巴细胞增值、小鼠脾淋巴细胞转化效应细胞及细胞黏附的影响，并用划痕标记染料示踪技术观察细胞间隙连接通讯功能；体内实验采用Lewis肺癌小鼠皮下移植模型、Lewis肺癌小鼠实验性肺癌转移模型和乌拉坦诱导的小鼠肺癌模型，评价皂角刺总黄酮对Lewis肺癌模型的预防和治疗作用。通过对实验小鼠的外观、体温、生存期和自主活动观察，评价实验小鼠的中医学体征的变化；通过肺组织HE染色法，观察肺癌组织病理学变化；通过检测血清指标：层粘连蛋白（LN）、血小板活化因子（PAF）、酪氨酸激酶B（TrkB）、酪氨酸激酶受体（TrkA）、小鼠免疫球蛋白G（IgG）的含量变化，评价皂角刺总黄酮对实验小鼠血清的影响；通过免疫组化法，观察皂角刺总黄酮对Cx43表达的影响，验证其抗肺癌的机制。体外实验结果显示：皂角刺总黄酮呈剂量相关性地抑制Lewis肺癌细胞增殖（IC50=146μg/ml），但对L929胚肺成纤维细胞增殖无显著影响。皂角刺总黄酮对细胞增殖影响较小的剂量可降低Lewis肺癌细胞粘附，并呈剂量相关性促进Lewis肺癌细胞间连接通讯，但对L929胚肺成纤维细胞粘附无影响。同时能提高体外培养的小鼠脾淋巴细胞增殖率及脾淋巴细胞的杀伤力。体内实验结果显示：与未治疗的Lewis肺癌皮下移植模型和Lewis肺癌实验性肺癌转移模型比较，皂角刺总黄酮高剂量和低剂量均可减缓肿瘤生长速度、阻止肿瘤转移、延长荷瘤小鼠存活时间。未注射乌拉坦的小鼠实验结束时肺部均无肿瘤发生，注射乌拉坦的模型小鼠肺部有数目和大小不等的肿瘤结节，皂角刺总黄酮高剂量组和低剂量组小鼠肺部肿瘤结节明显减少，部分小鼠未见肿瘤发生。与治疗组比较模型组小鼠体温下降、蜷缩少动；血清中LN、PAF、TrkB、TrkA水平升高，IgG含量降低。病理及免疫组化结果显示：与模型组的肺癌组织比较，皂角刺总黄酮高剂量组和低剂量组均能改善肺组织病理变化；而注射乌拉坦的模型小鼠肺部Cx43免疫组化显色密度明显降低，皂角刺总黄酮高剂量组和低剂量组均能增强致癌小鼠肺部的Cx43免疫组化显色密度。综上所述，皂角刺总黄酮对肺癌有确切的预防和治疗作用，其机制是增加免疫功能，增强细胞间连接通讯，恢复细胞间连接通讯的正常化，从而能阻止肿瘤生长和转移。
[Abstract]:Lung cancer is a kind of malignant tumor which is the most serious threat to life and human health. The incidence of lung cancer is the first of the malignant tumor and is increasing year by year. Therefore, it is of great significance to study the prevention and treatment of lung cancer. The traditional Chinese medicine for treating lung cancer mainly adopts the chemical medicine, but has the defects of toxic and side effect and multi-drug resistance in the treatment process. The traditional Chinese medicine has the advantages of small toxic and side effect in the treatment of the tumor, can improve the quality of life of the patient, and has the advantages of low cost of long-term administration, and the like, and is also an important source of the development of a novel anti-tumor medicament, Therefore, the screening of anti-tumor drugs of Chinese medicine has become a hot spot in the field of pharmacology. The total flavone of the spina Gleditsiae is a kind of flavonoid compound extracted from the dried echinax of the legume of the leguminous plant, and various exploration conclusions have shown that the proliferation of various tumor cells can be significantly affected by the total flavone of the spina Gleditsiae. The purpose of this study is to study the preventive effect and therapeutic effect of the total flavone of Spina Gleditsiae on the lung cancer cell and the lung cancer model, and to explore the possible implication of the new method. Mechanism. In vitro, MTT method was used to detect the effect of different concentrations of the total flavonoids of Spina Gleditsiae on the mouse lung cancer cells (Lewis), mouse embryonic lung fibroblasts (L929), the value-added of the spleen lymphocytes of the mouse, the transformation of the spleen and lymphocytes of the mouse, and the cells. The effects of cell adhesion on adhesion were observed and the intercellular communication function was observed with a scratch-labeled dye-tracer technique. Lewis lung cancer (Lewis lung cancer) mouse subcutaneous transplantation model, Lewis lung cancer mouse experimental lung cancer metastasis model and Uratin-induced small cell lung cancer were used in vivo. Study on the model of mouse lung cancer and the prevention of Lewis lung cancer model by using the total flavonoids of Spina Gleditsiae and the pathological changes of the lung cancer tissues were observed by means of a lung tissue HE staining method; and by detecting the serum index, the layer adhesion egg Changes of the content of white (LN), platelet-activating factor (PAF), tyrosine kinase B (TrkB), tyrosine kinase receptor (TrkA), and mouse immunoglobulin G (IgG), and the effect of the total flavonoids on the serum of the experimental mice. The effect of expression to verify its resistance to The results of in vitro experiments show that the total flavonoids of the Spina Gleditsiae inhibit the proliferation of Lewis lung cancer cells in a dose-dependent manner (IC50 = 146. mu.g/ ml), but the lung of the L929 embryo is fine. There was no significant effect on the proliferation of Lewis lung cancer cells. The dose of the total flavonoids of the Spina Gleditsiae to the proliferation of the cells decreased the adhesion of the Lewis lung cancer cells and promoted the intercellular communication between the Lewis lung cancer cells in a dose-dependent manner. has no effect on the adhesion of the dimension cells, and can improve the immunity rate and the spleen of the spleen lymphocytes of the mouse cultured in vitro. The results of in vivo experiments show that the high dose and low dose of the total flavonoids in the Lewis lung cancer can slow the growth rate of the tumor and prevent the metastasis of the tumor compared with the untreated Lewis lung cancer subcutaneous transplantation model and the Lewis lung cancer experimental lung cancer metastasis model. and the survival time of the tumor-bearing mice was prolonged. Some of the mice did not meet the tumorigenesis. Compared with the treatment group, the temperature of the mice in the model group was decreased and the volume of the mice was reduced; the serum LN, PAF, TrkB, TrkA water The results of pathology and immunohistochemistry showed that the high-dose group and low-dose group could improve the pathological changes of the lung tissues compared with the lung cancer tissues of the model group. 43. The color density of the immunohistochemical staining was significantly lower, and the high-dose group and the low-dose group of the total flavonoids of the Spina Gleditsiae can enhance the lung of the cancer-induced mice. In conclusion, the total flavonoids of Spina Gleditsiae have definite prevention and treatment effect on lung cancer. The mechanism is to increase the immune function, enhance the communication between the cells, and restore the communication between the cells.
【学位授予单位】：河南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965

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