【作者】 赵理想；

【导师】 陈思曾；

【作者基本信息】 福建医科大学 ， 外科学， 2010， 硕士

【摘要】 目的研究福莫特罗、吲哚美辛和罗红霉素对癌症恶病质小鼠的联合治疗作用并探讨其可能机制。方法于皮下接种小鼠结肠腺癌Colon26 (C26)细胞株于雄性Babl/c小鼠后9天建立癌症模型。72只小鼠按析因设计随机分为9组,每组8只;从第9天开始每天按分组情况分别给予不同的药物治疗。FM剂量1 mg/kg/d, IND剂量0.5 mg/kg/d,RM剂量50 mg/kg/d,以上药物全部采用腹腔注射给药,连用7天。每天监测小鼠进食量,每天称量小鼠体质量,从皮下可触及肿瘤后每天测量肿瘤体积。第16天处死各组小鼠,检测IL-6、TNF-α及白蛋白、甘油三酯、血糖水平。数据使用SPSS10.0统计软件包处理。结果:1癌性恶病质各组,皮下接种C26细胞4-5天后,肿瘤开始可以皮下触及,小鼠出现精神萎靡,活动减少。9天左右肿瘤长至平均约1cm3,小鼠开始出现全身毛色发暗且杂乱无序、活动迟缓、消瘦等表现,体质量亦出现下降,至第9天,全部接种肿瘤小鼠进入癌性恶病质状态。2腓肠肌重量实验结束时B组小鼠左侧腓肠肌重量降至A组的67%(P<0.05)。析因分析示:罗红霉素、吲哚美辛、福莫特罗用与不用在增加腓肠肌重量方面有差异(P<0.05),三种药物两两组合之间在增加左侧腓肠肌重量方面未发现有统计学意义(P>0.05),三中药物组合之间亦无交互作用(P>0.05),福莫特罗、吲哚美辛、罗红霉素三药联合组增加腓肠肌重量最多。3生化指标B组小鼠与A组小鼠相比,白蛋白、甘油三酯及血糖水平差异有显著性(P<0.05)。对于甘油三酯,福莫特罗、吲哚美辛和罗红霉素能使其水平降低(P<0.05),三种药物两两之间无交互作用(P>0.05),但三者之间存在交互作用(P<0.05),以三药联合用药组降低甘油三酯最多。对于血糖析因分析结果表明:福莫特罗、吲哚美辛、罗红霉素能使血糖升高(P<0.05),三种药物两两之间无交互作用(P>0.05),但三者之间存在交互作用(P<0.05)以三药联合用药组升高血糖最多。而对白蛋白析因分析结果表明:三药在升高白蛋白方面均没有统计学意义(P>0.05)。4血清细胞因子B组小鼠细胞因子TNF-α,IL-6和正常小鼠A组相比均有升高(p<0.05)。IL-6析因分析结果表明:吲哚美辛两水平间有差异(P<0.05),表明吲哚美辛能降低恶病质小鼠IL-6。而另外两药虽能降低IL-6,但无统计学意义(P>0.05),且药物间没有发现有交互作用(P>0.05)。TNF-α析因分析显示福莫特罗、吲哚美辛、罗红霉素主效应有统计学意义(P<0.001),表明三种药物单独应用都能降低TNF-α。福莫特罗与罗红霉素间有交互作用,且此三种药物间宜有交互作用,以三药联合用药组降低TNF-α最多。结论1. Colon26结肠腺癌诱导的癌症恶病质模型与人类癌症恶病质相似,是研究癌症恶病质发病机制及药物治疗实验的理想模型。2.癌症恶病质小鼠去瘤体质量下降、腓肠肌重量下降、严重的代谢紊乱,血清TNF-α, IL-6含量增高,提示代谢紊乱产生的TNF-α, IL-6等细胞因子与癌症恶病质的发生、发展密切相关,是癌症恶病质发生的一个重要机制。3.福莫特罗、吲哚美辛、罗红霉素在抗癌症恶病质代谢紊乱与组织消耗及降低血清中升高的TNF-α, IL-6方面都有着不同程度的作用。将促合成代谢药物(福莫特罗)和抗细胞因子药物(吲哚美辛、罗红霉素)联合应用治疗CC在某些方面可以产生相互的交互作用,治疗效果在某些方面优于单一应用。更多还原

【Abstract】 Objective To observe the effect of the Combinative Supplementation of Formoterol(FM), Indomethacin (IND)and Roxithromycin (RM) on cancer cachexia (CC) and to study the mechanism with an animal cancer cachetic model.Methods Male BALB/c mice bearing colon 26 adenocarcinoma for 9 days were served as models of cancer cachexia. The 72 models were divided randomly into nine treating groups basing on factorial design. Formoterol was used at a dose of 1 mg/kg/d, indomethacin was used at a dose of 0.5 mg/kg/d, roxithromycin was used at a dose of 50mg/kg/d. All of drugs were injected through intraperitoneal injection. FM, IND, FM and saline were given daily for 7days from the onset of cachexia to sacrifice. Physiological conditions, bodyweight and food intake were documented every day, tumor volume were documented every day after the tumors were palpable. Serum levels of cytokine and nutritional markers were detected 7 days after treatment. Data were expressed as means±SE. Statistical significance was determined by factorial design ANOVA and Student’s t-test using SPSS 10.0. P<0.05 was considered statistically significant.RESULT1 Cancer cachexia model Tumors were palpable in mice initially on day4 or 5 after inoculation of tumor cells, Mice becomes listless and less move. Tumor grew about 1cm3 after 9 days. Color pattern of mice lose luster and disorderly. Their action become sluggish, they become emaciation. Body weight of mice also begins to decrease. All mice get into cancer cachexia on the 9th day. Cachexia symptoms were observed, such as obviously poor physical activity, asthenia, piloerection, pelage lost gloss and sheded.2 left gastrocnemius weight When the experiment was finished, the left gastrocnemius weight of group B was cut down to 67% of group A(P<0.05). This experiment using three-factorial analysis showed that roxithromycin, indomethacin, formoterol can increase the left gastrocnemius alone(P<0.05), Any two of the three drugs have no interaction on elevating the left gastrocnemius weight. The three drugs combinative supplementation can mostly improve the weight of left gastrocemius.3. Biochemical indicators The levels of serum Glu and Alb in B group were obvious lower than group A (p<0.05), but the levels of TG were significant higher than group A (p<0.05). Factorial analysis indicate that FM, IND, RM can decrease the levels of TG(P<0.05). Any two of the three drugs have no interaction on decrease the levels of TG(P>0.05), but the three drugs have interaction on decrease the levels of TG(P<0.05). The three drugs combinative supplementation can mostly bring down the levels of TG. About alb factorial analysis indicate that FM, IND, RM can elevate the levels of GLU(P<0.05). Any two of the three drugs have no interaction on elevate the levels of glucose(P>0.05), but the three drugs have interaction on elevating the levels of GLU(P<0.05). The three drugs combinative supplementation can mostly improve the levels of GLU. Although Alb of groups with drugs treatment were raised, no statistical significance was detected in those groups (p >0.05). Factorial analysis indicate that FM, IND, RM have no statistical difference in elevating the levels of alb(P>0.05).4. Serum cytokines levels Comparing with normal mice, the cytokines TNF-αand IL-6 levels were raised significantly (p<0.05). After treatment with drugs, the results show that IND could decrease the levels of IL-6 (p<0.05), but FM and RM could not(p>0.05) and there are not interactions among FM , IND and RM in IL-6 level(sP>0.05). About TNF-αfactorial analysis indicate that FM, IND, RM can lower the levels of TNF-α(P<0.001. There are not only interactions between FM and RM in decreasing the levels of TNF-α, but also the three drugs have interactions in decreasing the levels of TNF-α. The three drugs combinative supplementation can mostly decrease the levels of TNF-α.Conclusion1. This experiment had established an animal model of cancer cachexia successfully that was very similar human’s by inoculating BALB/c mice with colon-26.It provides contribution for further studying cancer cachexia theory foundation and efficacy of drugs in treating cancer chachexia.2. The result of body weight decreasing, metabolic disorder and the levels of IL-6、TNF-αincreasing showed that mice in cancer cachexia condition appearance significantly metabolic disorder of glucose, lipoids and albumin. It may have correlation with cytokines. It showed that the participation of cytokines probably is one of important mechanism that is lead to happen cancer cachexia.3. Three drugs: FM, IND and RM had their own positive effects on CC mice. Drugs combination in tumor-bearing mice with cancer cachexia could produce synergistic action.更多还原