六月青石油醚萃取物对S180荷瘤小鼠抑瘤作用研究

【作者】 韦京辰；

【导师】 王乃平； 黄仁彬；

【作者基本信息】 广西医科大学 ， 药理学， 2007， 硕士

【摘要】 从中药寻找抗肿瘤有效成分具有广阔的前景。六月青在广西分布广泛。药理实验证实其具有抗病毒作用,一些初步的实验证实其有一定的抗肿瘤活性。但在现阶段,对六月青系统的抗肿瘤活性及机理研究几乎处于空白,本课题对六月青提取物进行抗肿瘤实验及相关研究,为六月青的进一步研究及开发利用提供科学依据。目的:研究六月青石油醚萃取物（petroleum ether extract of LYQ,LYQPE）对S₁₈₀荷瘤小鼠的抑瘤作用及其作用机制。方法:①建立S₁₈₀荷瘤小鼠腋下移植瘤模型:取小鼠50只,右腋皮下接种S₁₈₀腹水肿瘤细胞0.2 ml,24 h后随机分为5组:模型组,阳性对照组（CTX）,以LYQPE灌胃的高剂量、中剂量、低剂量药物组。每组10只小鼠,接种24 h后开始给药,给药10天。停药24 h后眼球采血、处死,摘取瘤组织称重,计算抑瘤率。②抗肿瘤免疫学:摘取S₁₈₀荷瘤小鼠胸腺和脾脏,计算胸腺指数和脾脏指数;制备脾淋巴细胞悬液,用Con A刺激脾淋巴细胞增殖,MTT法检测脾淋巴细胞转化功能;制备腹腔巨噬细胞（MΦ）悬液,中性红法检测腹腔巨噬细胞吞噬功能;用ELISA法检测血清肿瘤细胞坏死因子（TNF-α）和白细胞介素-2（IL-2）的含量。③抗氧化作用:用比色法测S₁₈₀小鼠血浆丙二醛（MDA）和超氧化物歧化酶（SOD）含量。④瘤组织病理学改变:取瘤组织块,固定,HE染色观察瘤组织病理学改变及免疫组化法观察P₅₃蛋白的表达。结果:①LYQPE能抑制S₁₈₀荷瘤小鼠移植性肿瘤,与模型组比较有显著性差异（P<0.01或P<0.05）,高剂量组、中剂量组抑瘤率高于30%。②LYQPE能增高胸腺指数,高剂量组与模型组比较显著差异（P<0.05）,但对脾指数无明显作用（P>0.05）;能促进脾淋巴细胞增殖,高剂量组、中剂量组与模型组比较有显著性差异（P<0.01）;能增高MΦ吞噬功能,高剂量组、中剂量组与模型组比较有显著性差异（P<0.01 or P<0.05）。LYQPE增高血清TNF-α含量,高剂量组、中剂量组与模型组比较有显著性差异（P<0.01 orP<0.05）;能增高IL-2的含量,高剂量组、中剂量组、低剂量组与模型组比较有显著性差异（P<0.01）。③LYQPE能降低血浆MDA含量,升高SOD含量,高剂量、中剂量、低剂量组与模型组比较有显著性差异（P<0.05或P<0.01）。④HE染色可见LYQPE药物组瘤组织有病理学改变,高剂量、中剂量组瘤组织坏死严重。LYQPE能降低瘤组织中P₅₃蛋白表达,高剂量、中剂量、低剂量组均有显著性差异（P<0.05）。结论:LYQPE对S₁₈₀荷瘤小鼠有明显的抑瘤作用,其可能机制与改善小鼠免疫功能、清除自由基、增强机体抗氧化功能和诱导肿瘤细胞凋亡有关。更多还原

【Abstract】 It has broad prospect from traditional Chinese medicine to find active ingredients for antitumor.Liuyueqing is widely distributed in Guangxi. Pharmacological research confirme that Liuyueqing has antivirus effect.Some preliminary experiments prove Liuyueqing extracts have antitumor activity. However,systematic study of antitumor activity on Liuyueqing is almost blank at the present.In this experiment,we study the antitumor activity of extract from Liuyueqing to produce the scientific evidence for its further research and exploitation.Objective:To study the inhibitory effect and its mechanism of LYQPE on transplanted S₁₈₀sarcoma model in mice.Methods:（1）Transplanted S₁₈₀sarcoma in mice was used as a model:50 mice were inoculated subcutaneously with 0.2 ml of S₁₈₀sarcoma cell in right axilla.24 hours afher inoculation,these mice were randomly divided into five groups:tumor model group,CTX positive control group,petroleum ether extract of LYQ（LYQPE）high-dose group,middle-dose group and low-dose group（n= 10）.All mice were treated with drug after 24 h after inoculation for 10 days. Blood was collected,and mice were sacrificed 24 h after last dose of drug. Tumor tissues were weighed,and inhibitory rate of tumor was calculated.（2） Effect of LYQPE on immunological function:thymus and spleen of mice of transplanted S₁₈₀sarcoma were culled and weighed.Index of thymus and spleen were calculated.Spleen lymphocyte suspension was prepared and stimulated by Con A.Proliferated ability of lymphocyte was assayed by MTT method. Macrophage suspension was prepared and it’s the phagocytized ability of macrophage was assayed by neutral red method.The content of tumor necrosis factor-a（TNF-α）and interleukin-2 were assayed by ELISA.（3）Anti-oxidative effect of LYQPE:The content of maleicdialdehyde（MDA）and superoxide dismutase（SOD）were assayed by colorimetry.（4）Effect of LQYPE on pathological change in S₁₈₀tumor tissue:tumor tissue was fixed by formaldehyde,and dyed by HE stain and pathological changes were observed by light microscope,and P₅₃protein expression in tumor tissue was determined by immunohistochemical method.Results.（1）LYQPE could inhibit the growth of transplanted S₁₈₀sarcoma in mice.Inhibitory rates in high-dose group;middle- dose group and low-dose group were were significantly different from that in model group（P＜0.01 or P＜0.05）.The inhibitory rates of high-dose group and middle-dose group were more than 30%.（2）LYQPE could increase the indexes of thymus,which in high-dose group was increased significantly（P＜0.05）,but LYQPE had not influence on the indexes of spleen of mice with S₁₈₀tumor（P＞0.05）.LYQPE could stimulate lymphocyte proliferation,which in high-dose group and middle-dose group were significantly different from that in model group （P＜0.01）.LYQPE could increase phagocytized ability of macrophage,which in high-dose group and middle-dose group was significantly different from model group（P＜0.01 or P＜0.05）.LYQPE could increase the content of tumor necrosis factor-a,which in high-dose group and middle-dose group were significantly different from that in model group（P＜0.01 or P＜0.05）.LYQPE could increase the content of interleukin-2,which in high-dose group,middle-dose group and low-dose group were significantly different from that in model group（P＜0.01）. （4）LYQPE group could decrease the content of MDA,and increase the activity of SOD,which in high-dose group,middle- dose group and low-dose group were significantly different from that in model group（P＜0.01 or P＜0.05）.（5） Acellular necrosis of tumor tissue was more serious in high-dose group and middle-dose group that in model group.LYQPE could decrease mt P₅₃protein expression,which in igh-dose group,middle-dose group and low-dose group were significantly different from model group（P＜0.05）.Conclusions:LYQPE can inhibit the growth of transplanted S₁₈₀sarcoma in mice,and the increases in the immunological function,antioxidative ability and induced apoptosis of tumor cell by LYQPE were the possible action mechanism.更多还原