加味旋覆代赭汤对非酸反流所致慢性食管病变大鼠肠化及CDX2、DLK1表达的影响

Effect of Modified Xuanfu Daizhe Decoction on Intestinal Metaplasia and Expression of CDX2 and DLK1 in Rats with Chronic Esophageal Mucosal Injury Induced by Non-acid Reflux

目的:采用十二指肠-食管吻合术(胃全切)建立大鼠十二指肠-食管非酸反流模型,观察加味旋覆代赭汤对模型食管肠化及CDX2、DLK1表达的影响。方法:成年雄性Wistar大鼠80只,随机分为假手术组、模型组、加味旋覆代赭汤组及铝碳酸镁片(达喜)组,每组20只。除假手术组外,其余三组均采用十二指肠-食管吻合术(胃全切)建立大鼠十二指肠-食管非酸反流模型,建模25周时,加味旋覆代赭汤组经口灌服加味旋覆代赭汤(相当于30 g生药/kg),每日1次,连续3周;达喜组经口灌服达喜药液(300 mg/kg),每日1次,连续3周;模型组给予同体积蒸馏水经口灌服,每日1次,连续3周。治疗3周后,连同假手术组,共4组动物,麻醉状态下打开胸腹腔,剪取食管下段及吻合段,纵行剖开,进行大体观察,对食管下端紧邻吻合口处组织行HE染色,行食管病理组织学检查并评分,另采用免疫组化法进行CDX2及Notch信号分子-DLK1染色,观察其表达情况。结果:大体观察发现模型组食管下段显著增粗,纵行剖开食管,可见下端食管显著增厚,食管下段黏膜呈广泛显著不规则隆起,外观颜色呈白斑样改变;加味旋覆代赭汤组食管大体病理较模型组显著减轻,达喜组食管大体改变较模型组无显著差别。模型组轻、中、重度食管炎发生率分别为2/15(13.33%)、3/15(20.00%)、10/15(66.67%);加味旋覆代赭汤组分别为11/17(64.70%)、3/17(17.65%)、3/17(17.65%),与模型组比较炎症程度显著降低(P<0.01),达喜组分别为3/16(18.75%)、4/16(25.00%)、9/16(56.25%),与模型组差异无显著性(P>0.05)。加味旋覆代赭汤组的Barrett食管发生率为2/17(11.8%),显著低于模型组的7/15(46.7%),差异有统计学意义(P<0.05),达喜组为7/16(43.8%),与模型组差异无统计学意义(P>0.05)。加味旋覆代赭汤组的CDX2、DLK1表达水平均显著低于模型组(P<0.01),而达喜组与模型组比较差异无统计学意义(P>0.05)。结论:加味旋覆代赭汤可通过抑制CDX2及Notch信号分子DLK1表达来抑制肠化,减少非酸反流所致Barrett食管的发生。

Objective Duodenal-esophageal anastomosis(total gastrectomy)was used to establish a rat model of duodenal-esophageal non-acid reflux.The effects of Modified Xuanfu Daizhe Decoction(MXDD)on esophageal intestinalization,CDX2 and DLK1 expression were observed.Methods Eighty adult male Wistar rats were randomly divided into sham-operated group,model group,MXDD group and aluminum magnesium carbonate tablet(Daxi)group,20 rats in each group.Except for the sham-operated group,the other three groups used duodenoesophageal anastomosis(total gastrectomy)to establish the rat duodenal-esophageal non-acid reflux model.At 25 weeks,the rats in MXDD group were given MXDD(equivalent to 30 g crude drug/kg),once a day for 3 weeks;the rats in Daxi group were orally given Daxi liquid(300 mg/kg),once a day for 3 weeks;the rats in model group were given distilled water of the same volume,once a day for 3 weeks.After 3 weeks of treatment,the thoraco-abdominal cavities of four groups of animals were opened under anesthesia,and the lower esophageal segment and anastomotic segment were cut,and longitudinal dissection was made for general observation.The tissue adjacent to anastomotic opening at the lower end of the esophagus was stained with HE to observe the pathological changes of esophagus.The histopathology of the esophagus was examined and scored.In addition,CDX2 and Notch signal molecule DLK1 were stained by immunohistochemistry to observe their expression.Results Gross observation showed that the lower esophagus of the model group was significantly thicker,the lower esophagus was significantly thicker,the lower esophagus mucosa was widely irregular bulge,and the appearance color was white spot.The gross pathology of esophagus in MXDD group was significantly lighter than that in model group,while that in Daxi group was slightly lighter than that in model group.The incidence of mild,moderate and severe esophagitis in the model group was 2/15(13.33%),3/15(20.00%)and 10/15(66.67%)respectively,and 11/17(64.70%),3/17(17.65%)and 3/17(17.65%)in MXDD group were significantly lower than those in the model group(P<0.05),and 3/16(18.75%),4/16(25%),9/16(56.25%)respectively.There was no significant difference in the model group(P>0.05).The incidence of Barrett esophagus was 7/15(46.7%)in the model group and 2/17(11.8%)in the MXDD group,which was significantly lower than that in the model group(P<0.05),and 7/16(43.8%)in the Daxi group(P>0.05).The expression of CDX2 and DLK1 in the MXDD group was significantly lower than that in the model group(P<0.05),but there was no significant difference between the Daxi group and the model group(P>0.05).Conclusion MXDD can inhibit intestinal metaplasia and reduce Barrett esophagogenesis caused by non-acid reflux by inhibiting the expression of CDX2 and Notch signaling molecule-DLK1.