四神丸对伊立替康所致的小鼠迟发性腹泻防治作用的实验研究

发布时间：2018-03-29 19:42

  本文选题：四神丸　切入点：伊立替康　出处：《南京中医药大学》2017年硕士论文

【摘要】：目的:迟发性腹泻为伊立替康(Irinotecan,CPT-11)主要剂量限制性毒性,严重影响患者的生活质量,甚至导致治疗中断,影响患者的疗效。目前依然缺乏较满意的防治方法。在本项研究中,建立小鼠CPT-11所致迟发性腹泻模型,研究四神丸各剂量组对CPT-11引起的迟发性腹泻的防治作用,并探讨其可能的机制。方法:1)迟发性腹泻模型的建立、分组50只健康雄性ICR小鼠被随机分为5组:腹泻模型组、四神丸低(0.6g·kg-1)、中(1.2g·kg-1)、高(2.4g·kg-1)剂量组、正常对照组,每组10只。腹泻模型组及四神丸各剂量组均尾静脉注射CPT-11(35mg·kg-1·d-1),每日1次,连续4日;正常对照组小鼠尾静脉被注射同等体积生理盐水。四神丸各剂量组于造模前一天灌胃给药,每日1次,连续7日,腹泻模型组及正常对照组小鼠被给予同等体积双蒸水灌胃。2)每日观察、记录小鼠迟发性腹泻发生的情况、饮食及精神活动状态等。3)样本采集小鼠末次灌胃给药24h后,腹腔注射水合氯醛麻醉,腹主动脉取血;断颈后处死,取结肠组织及盲肠内容物。4)样本的检测通过酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测小鼠血样中的白介素 1β(Interleukin-1β,IL-1β)、肿瘤坏死因子 α(tumor necrosis factor-α,TNF-α)的含量;比色法检测盲肠内容物β-葡萄糖醛酸苷酶(β-glucuronidase,β-G)的表达水平;盲肠内容物大肠杆菌培养,计数菌落数;结肠组织被固定、切片、HE染色后,于显微镜下观察结肠粘膜的结构和形态。结果:1)迟发性腹泻的发生情况:小鼠尾静脉注射CPT-11后,于24h后出现迟发性腹泻,腹泻模型组小鼠发生迟发性腹泻率为100%,其中重度腹泻发生率为80%;四神丸中、高剂量组重度腹泻发生率分别为40%;与腹泻模型组比较:四神丸中、高剂量组腹泻程度明显减轻(P=0.021,P=0.019)。2)结肠粘膜的损伤情况腹泻模型组小鼠5-6级粘膜损伤发生率为70%,而四神丸中、高剂量组均为30%,与腹泻模型组比较,差异有统计学意义(P0.05)。3)小鼠血清IL-1β的变化情况四神丸中高剂量组血清IL-1β分别为:(72.61 土 10.15)/g·L-1,(71.86 土 8.78)/g·L-1较腹泻模型组(116.57 土 17.16)/g·L-1的水平显著降低(P0.01)。4)小鼠血清TNF-α的变化情况四神丸中高剂量组血清TNF-α分别为:(71.50 土 3.39)/g·L-1,(69.43 土 6.23)/g·L-1较腹泻模型组(109.50 土 4.21)/g·L-1的水平显著降低(P0.01)。5)小鼠肠道β-G的表达水平四神丸中、高剂量组肠道β-G分别为:(1.109 土 0.055)U,(1.100 土 0.063)U;较腹泻模型组(1.438 土 0.022)U的表达显著降低(P0.01)。6)小鼠盲肠内大肠杆菌菌落数:四神丸低中高剂量组分别为(29.5 土 2.2)×1013/g,(30.4 土 3.5)×1013/g,(31.4 土2.7)×1013/g;腹泻模型组(31.4 土 3.2)×1013/g;正常对照组(31.2 土 2.9)×1013/g。各组之间比较,菌落数无明显差异(P0.05)。结论:四神丸对CPT-11所致小鼠迟发性腹泻有一定的预防和减缓作用,其机制可能与减少肠道β-G的含量,并降低血清中IL-1β、TNF-α的水平,从而减轻CPT-11对肠道黏膜的损伤有关。
[Abstract]:Objective: delayed diarrhea for irinotecan (Irinotecan, CPT-11) the main dose limiting toxicity, seriously affect the patient's quality of life, and even lead to the interruption of treatment, therapeutic efficacy. There is currently no satisfactory treatment method. In this study, mice CPT-11 induced delayed diarrhea model, prevention and control to study the effect of Sishen pill each dose group of CPT-11 induced delayed diarrhea, and to explore its possible mechanism. Methods: 1) the establishment of delayed diarrhea model group, 50 healthy male ICR mice were randomly divided into 5 groups: diarrhea model group, Sishen pill low (0.6g - kg-1), in (1.2g - kg-1) (2.4G, kg-1), high dose group, normal control group, 10 rats in each group. The diarrhea model group and Sishen pill groups were intravenous injection of CPT-11 (35mg kg-1 D-1), 1 times a day for 4 days; the normal control group by tail vein of mice injected with the same volume of physiological four saline. God pill groups one day before the rats administered orally, 1 times daily for 7 consecutive days, diarrhea model group and normal control group mice were given the same volume of double distilled water.2) observed daily, delayed diarrhea cases were recorded, diet and mental activity status like.3) the last acquisition of mice administered 24h after intraperitoneal injection of chloral hydrate anesthesia, abdominal aortic blood; broken neck after death, the colon and cecum.4) samples was detected by enzyme-linked immunosorbent assay (enzyme-linked immunosorbent assay, ELISA) were detected in blood interleukin 1 beta (Interleukin-1 beta, beta IL-1), tumor necrosis factor alpha (tumor alpha factor- alpha necrosis, TNF-) content; detection of cecal contents of beta glucuronidase assay (beta -glucuronidase and beta -G) expression level; cecal contents of Escherichia coli culture, counting the number of colony; colon tissue Is fixed, sliced, HE stained, and observed the structure and morphology of colonic mucosa. Results: 1) delayed diarrhea: tail vein of mice after injection of CPT-11, appeared in 24h after delayed diarrhea, diarrhea model mice delayed diarrhea rate was 100%, and the severe diarrhea the incidence rate was 80%; shishenwan, high dose group and severe diarrhea incidence was 40%; compared with the model group: shishenwan diarrhea, diarrhea in high dose group reduced significantly (P=0.021, P=0.019).2) in colonic mucosal injury diarrhea mice model 5-6 mucosal injury incidence rate was 70%, and four pills in high dose group was 30%, compared with the diarrhea model group, the difference was statistically significant (P0.05).3) of Sishen pill mice serum IL-1 beta in high dose group serum IL-1 beta respectively: (72.61 + 10.15) /g - L-1 (71.86, /g, L-1 8.78) compared with the model of diarrhea group (1 16.57 鍦，

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