表没食子儿茶素没食子酸酯、β-羟基-β甲基丁酸盐联合治疗癌症恶病质实验性研究

发布时间：2018-05-11 10:53

本文选题：恶病质 + 表没食子儿茶素没食子酸酯　；参考：《福建医科大学》2013年硕士论文

【摘要】：目的观察表没食子儿茶素没食子酸酯(EGCG)、β-羟基-β甲基丁酸盐(HMB)联合应用对小鼠癌症恶病质的治疗作用。并初步探讨药物发挥作用的可能机制。方法接种小鼠结肠腺癌Colon26(C26)细胞的雄性BALB/C小鼠前腋窝皮下，9天后癌症恶病质模型基本建立。40只小鼠随机分为5组：A组空白对照组、B组荷瘤安慰剂（生理盐水）对照组、C组EGCG治疗组、D组HMB治疗组、E组联合治疗组,每组8只。每天按时监测小鼠食物摄入量，小鼠体质量及肿瘤体积。第16天，记录小鼠右侧腓肠肌重量和去瘤体重，检测血生化指标、血清白细胞介素-6(IL-6)、肿瘤坏死因子-ɑ(TNF-α)、C-反应性蛋白（CRP）水平，并用免疫组织化学法检测肿瘤组织核因子-κB(NF-κB)表达、荧光定量PCR腓肠肌MAFbx及MURF-1基因的表达。结果 1癌症恶病质模型的建立后四组小鼠皮下接种colon-26细胞4~5天后，肿瘤开始可以皮下触及。1周后长至约0.5cm3大小，小鼠开始出现毛色发暗、竖立、脱落和活动迟缓，活动减少、虚弱等表现。至第9天，，所有荷瘤小鼠的衰弱表现更加明显，且与正常对照组相比体质量显著下降(p＜0.05)，即进入恶病质状态。 2体重变化及摄食量情况接种初始几天各组间差异并无显著性，荷瘤小鼠体质量在第12天开始下降，且显著低于正常小鼠体质量（P＜0.05），到第16天降至最低，各组终末体质量各组荷瘤小鼠组间差异无统计学意义（P＞0.05）。但去瘤体质量荷瘤小鼠均显著低于正常小鼠（P＜0.05）。各组间食物摄食量各组间无明显差异（P＞0.05）。 3肿瘤体积变化荷瘤小鼠肿瘤从第5天开始可触及，肿瘤体积从第9天开始增大迅速，且EGCG治疗组、HMB治疗组、联合治疗组较安慰剂治疗组增长缓慢。至第16天各治疗组显著小于安慰剂治疗组。 4腓肠肌质量腓肠肌质量EGCG治疗组、HMB治疗组、联合治疗组、安慰剂治疗组与正常对照组间，EGCG治疗组和HMB治疗组与联合治疗组间的差异有统计学意义（P＜0.05），EGCG治疗组与HMB治疗组间的差异有统计学意义（P＜0.05）。 5血清生化指标的比较TP、ALB、Glu、TG水平EGCG治疗组、HMB治疗组、联合治疗组、安慰剂治疗组与正常对照组间，EGCG治疗组、HMB治疗组、联合治疗组与安慰剂治疗组间的差异有统计学意义（P＜0.05）。 6细胞因子(TNF-α、IL-6、CRP) EGCG治疗组、HMB治疗组、联合治疗组、安慰剂治疗组与正常对照组间，EGCG治疗组、HMB治疗组，联合治疗组与安慰剂治疗组间的差异均有统计学意义（P＜0.05）。EGCG治疗组、HMB治疗组和联合治疗组间的差异均有统计学意义（P＜0.05）。 7NF-κB表达水平的比较EGCG治疗组、HMB治疗组，联合治疗组与恶病质对照组安慰剂治疗组间的差异均有统计学意义（P＜0.05）。EGCG治疗组、HMB治疗组和联合治疗组间的差异均有统计学意义（P＜0.05）。 8MAFbx及MuRF-1的表达水平比较EGCG治疗组、HMB治疗组、联合治疗组、安慰剂治疗组与正常对照组间，EGCG治疗组、HMB治疗组，联合治疗组与安慰剂治疗组间的差异均有统计学意义(P＜0.05）。EGCG治疗组、HMB治疗组和联合治疗组间的差异均有统计学意义（P＜0.05）。结论 1EGCG和HMB联合疗效优于单一用药。 2可能通过抑制NF-κB，改善骨骼肌代谢，治疗癌症恶病质。
[Abstract]:objective
To observe the therapeutic effect of the combination of epigallocatechin gallate (EGCG) and beta hydroxy beta methylbutyrate (HMB) on cancer cachexia in mice, and to explore the possible mechanism of the effect of the drug.
Method
The male BALB/C mice of the colon adenocarcinoma Colon26 (C26) cells were subcutaneously inoculated into the anterior armpit of the male BALB/C mice. After 9 days, the cancer cachexia model was randomly divided into 5 groups: the A group blank control group, the B group, the placebo (physiological saline) control group, the C group EGCG treatment group, the D group HMB treatment group, and the E group combined treatment group, 8 in each group. Every day was monitored on time. Mice food intake, body mass and tumor volume. Sixteenth days, the weight of the right gastrocnemius and tumor weight were recorded, blood biochemical indexes, serum interleukin -6 (IL-6), tumor necrosis factor (TNF- alpha), C- reactive protein (CRP), and the expression of nuclear factor kappa B (NF- kappa B) in tumor tissues were detected by immunohistochemistry. The expression of MAFbx and MURF-1 genes in gastrocnemius muscle was quantified by PCR.
Result
1 after the establishment of the 1 cachexia model, four groups of mice were subcutaneously inoculated with Colon-26 cells for 4~5 days. The tumor began to grow subcutaneously to about 0.5cm3 after.1 weeks, and the mice began to appear darkening, erect, shedding, slow activity, activity decrease, weakness and so on. To ninth days, the decline of all tumor bearing mice was more obvious and positive. The body weight of the control group was significantly lower than that of the control group (P < 0.05), that is, entering cachexia.
2 the difference of weight change and feeding amount was not significant in the first few days. The body mass of the tumor bearing mice began to decrease at the beginning of twelfth days, and significantly lower than normal mice body mass (P < 0.05), and to the lowest level in sixteenth days. There was no significant difference between the groups of all groups of end body mass bearing tumor bearing mice (P > 0.05). Tumor mice were significantly lower than normal mice (P < 0.05). There was no significant difference in food intake among groups (P > 0.05).
3 tumor volume changes were palpable in the tumor bearing mice from fifth days. The tumor volume began to increase rapidly from the ninth day, and the EGCG treatment group, the HMB treatment group, the combined treatment group increased slowly compared with the placebo group. To sixteenth days, the treatment group was significantly smaller than the placebo treatment group.
4 gastrocnemius mass of gastrocnemius mass EGCG treatment group, HMB treatment group, combined treatment group, placebo group and normal control group, EGCG treatment group and HMB treatment group and the joint treatment group differences were statistically significant (P < 0.05), EGCG treatment group and HMB treatment group differences were statistically significant (P < 0.05).
5 Comparison of serum biochemical indexes: TP, ALB, Glu, TG level EGCG treatment group, HMB treatment group, combined treatment group, placebo treatment group and normal control group, EGCG treatment group, HMB treatment group, the difference between the combined treatment group and the placebo group was statistically significant (P < 0.05).
6 cell factor (TNF-, IL-6, CRP) EGCG treatment group, HMB treatment group, combined treatment group, placebo treatment group and normal control group, EGCG treatment group, HMB treatment group, the difference between the combined treatment group and the placebo treatment group were statistically significant (P < 0.05).EGCG treatment group, HMB treatment group and the joint treatment group differences were statistically significant ( P < 0.05).
The comparison of the expression level of 7NF- kappa B was statistically significant between the EGCG treatment group, the HMB treatment group, the combined treatment group and the cachexia control group (P < 0.05).EGCG treatment group, and the difference between the HMB treatment group and the combined treatment group had statistical significance (P < 0.05).
The expression level of 8MAFbx and MuRF-1 was compared between the EGCG treatment group, the HMB treatment group, the combined treatment group, the placebo treatment group and the normal control group, the EGCG treatment group, the HMB treatment group, the difference between the combined treatment group and the placebo group were statistically significant (P < 0.05).EGCG treatment group, the difference between the HMB treatment group and the combined treatment group was statistically significant. Significance (P < 0.05).
conclusion
The combination of 1EGCG and HMB was better than a single drug.
2, it is possible to improve the metabolism of skeletal muscle and cure cancer cachexia by inhibiting NF- kappa B.

【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R73-3

【参考文献】