【摘要】 目的探讨生姜泻心汤对伊立替康(CPT-11)化疗后大鼠肠黏膜及肠功能细胞的影响。方法健康雄性SD大鼠24只,按随机数字表法分为正常对照组、CPT-11组、中药加CPT-11组,每组8只。CPT-11组、中药加CPT-11组于实验第4日尾静脉注射CPT-11150 mg/kg,每日1次,连续2日,复制迟发性腹泻模型,正常对照组尾静脉注射等体积生理盐水。中药加CPT-11组于实验第1日,用生姜泻心汤(2 g/mL)灌胃给药,每只大鼠10 g/kg,连续用药9日,CPT-11组、正常对照组去离子水灌胃。于48、60、72、84、96及108 h时观察记录大鼠腹泻情况,计算腹泻发生率,同时参考Akinobu K方法进行腹泻评分。实验第10日处死大鼠,取回肠、盲肠及结肠组织,10%福尔马林液固定。行HE染色,在光镜下根据chiu标准对肠黏膜损伤分级。PAS染色观察肠杯状细胞、潘氏细胞表达;免疫组化CgA染色观察肠内分泌细胞形态,用Image-Pro-Plus 6.0对阳性细胞进行计数和累计光密度分析。结果正常对照组大鼠各时间点均无腹泻发生。CPT-11组大鼠48、60、72、84、96及108 h腹泻发生率为75.0%(6/8)、100.0%(8/8)、100.0%(8/8)、87.5%(7/8)、75.0%(6/8)及75.0%(6/8)。中药加CPT-11组腹泻发生率48 h为25.0%(2/8),60h为50.0%(4/8),72 h为12.5%(1/8),84 h后腹泻发生率为0.0%(0/8)。与本组60 h比较,CPT-11组48、84、96、108 h及中药加CPT-11组48、72、84、96、108 h腹泻评分明显降低(P<0.05,P<0.01);与本组72 h比较,CPT-11组84、96及108 h腹泻评分明显降低(P<0.05,P<0.01)。与正常对照组比较,CPT-11组各时间点腹泻评分升高(P<0.01),回肠、盲肠及结肠黏膜损伤分级升高(P<0.05,P<0.01),回肠、盲肠黏膜上皮杯状细胞表达明显减少(P<0.05),潘氏细胞数量及表达降低(P<0.01),内分泌细胞在回肠内表达增高,盲肠及结肠内表达减少(P<0.01);与CPT-11组比较,中药加CPT-11组腹泻评分明显降低(P<0.05,P<0.01),回肠、盲肠黏膜损伤分级降低(P<0.05,P<0.01),回肠、盲肠及结肠黏膜上皮杯状细胞表达均明显增加(P<0.05,P<0.01),回肠潘氏细胞数量及表达升高(P<0.05),内分泌细胞在盲肠及结肠内表达增加(P<0.05,P<0.01)。结论生姜泻心汤通过改善CPT-11化疗所致肠黏膜及肠功能细胞的凋亡、坏死,从而对CPT-11所致迟发性腹泻具有预防和治疗作用。更多还原

【Abstract】 Objective To explore the effect of Shengjiang Xiexin Decoction(SXD) on the intestinal mucosal and functional cells of rats after irinotecan(CPT-11) chemotherapy.Methods Totally 24 healthy Sprague-Dawley(SD) male rats were divided into three groups,the normal control group,the CPT-11 group,the SXD combined CPT-11 group according to random digit table,8 in each group.CPT-11 was injected at the daily dose of 150 mg/kg to rats in the CPT-11 group and the SXD combined CPT-11 group from the caudal vein on the 4th day,once daily for 2 successive days to duplicate delayed diarrhea model.Equal volume of normal saline was injected to rats in the normal control group from the caudal vein.SXD at 2 g/mL(10 g/kg body weight) was administered to rats in the SXD combined CPT-11 group by gastrogavage for 9 successive days.Deionized water was administered to rats in the CPT-11 group and the normal control group.Diarrhea was observed at 48,60,72,84,96,and 108 h to calculate the incidence rate of diarrhea.Meanwhile,scoring for diarrhea was performed by referring methods of Akinobu Kurita.Rats were killed on day 10,ileum,cecum,and colon tissues were collected and fixed in 10%formalin solution.HE staining was performed.Intestinal mucosa injuries were graded under light microscope according to the criterion of Chiu’s score.The expressions of goblet cells and Paneth cells were observed by PAS stain.Enteroendocrine cells were observed by immunohistochemical CgA staining.Positive cells were counted and cumulative optical density(IOD) analyzed by Image-Pro-Plus 6.0.Results No diarrhea occurred in rats of the normal control group at each time point.The incidence rate of diarrhea was75.0%(6/8)at 48 h,100.0%(8/8) at 60 h,100.0%(8/8) at 72 h,87.5%(7/8) at 84 h,75.0%(6/8)at 96 h,and 75.0%(6/8) at 108 h in the CPT-11 group.The incidence rate of diarrhea was 25.0%(2/8) at48 h,50.0%(4/8) at 60 h,12.5%(1/8) at 72 h,0.0%(0/8) at 84 h in the SXD combined CPT-11 group.Compared with the same group at 60 h,scores for diarrhea at 48,84,96,and 108 h obviously decreased in the CPT-11 group,and scores for diarrhea at 48,72,84,96,and 108 h obviously decreased in the SXD combined CPT-11 group(P<0.05,P<0.01).Compared with the same group at 72 h,scores for diarrhea at 84,96,and 108 h obviously decreased in the CPT-11 group(P <0.05,P <0.01).Compared with the normal control group,scores for diarrhea increased in the CPT-11 group at each time point(P <0.01);grading of ileum,cecum,and colon mucosal tissues increased(P<0.05,P<0.01);expressions of ileum and cecum mucosal epithelial goblet cells obviously decreased(P <0.05);the number and expressions of ileum and cecum mucosal epithelial Paneth cells increased(P <0.01).Expressions of ilium endocrine cells increased,while those of cecum and colon endocrine cells decreased in the CPT-11 group(P <0.01).Compared with the CPT-11 group,scores for diarrhea were obviously lowered(P <0.05,P <0.01),grading of ileum,and cecum mucosal tissues decreased(P <0.05,P <0.01);expressions of ileum,cecum,and colon mucosal epithelial goblet cells obviously increased(P <0.05,P <0.01);the number and expressions of ileum cecum mucosal epithelial Paneth cells increased(P <0.05) expressions of cecum and colon endocrine cells increased(P<0.05,P <0.01) in the SXD combined CPT-11 group.Conclusion SXD played roles in preventing and treating CPT-11 induced delayed diarrhea by improving CPT-11 chemotherapy induced apoptosis and necrosis of intestinal mucosal and functional cells.更多还原