辛开苦降法对反流性食管炎模型大鼠食道下段组织iNOS蛋白表达的影响

Impacts of Xinkaikujiang Therapy on iNOS Protein Expression in the Lower Esophageal Tissue of the Rat Models of Reflux Esophagitis

目的探讨辛开苦降法对反流性食管炎(RE)模型大鼠食道下段组织诱导型一氧化氮合酶(iNOS)蛋白表达的影响。方法将98只成年雄性wistar大鼠随机分为正常组(10只)和造模组(88只)。用改良部分贲门肌切开术+外置幽门半结扎术进行造模,造模后去除死亡大鼠和虚弱大鼠,选取恢复较好的大鼠60只将其随机分为模型组,中药低、中、高剂量组,中西药混合组、西药组,每组各10只。正常组不予处理,各组分别给予相应的药物灌胃,模型组给予0.9%生理盐水,中药低、中、高剂量组分别给予不同浓度的以辛开苦降法为主的半夏泻心汤加减化裁,西药组予含多潘立酮、铝碳酸镁、雷贝拉唑钠的混合药液,中西药混合组予中药中剂量组及西药组的混合药液。依据治疗时间不同,每组又分为7 d、14 d两个亚组,每个亚组5只大鼠。分别于治疗第7天和第14天取材,采用免疫组化和western-blot法,检测各组食管下段黏膜iNOS的表达水平。结果模型组与正常组对比,大鼠食道下端iNOS表达增加(P<0.05),治疗7天后,各组与模型组比较,iNOS表达没有显著差异(P>0.05);治疗14天后,中药高剂量组、中西药混合组、西药组与模型组比较,iNOS表达均有不同程度减少,差异有统计学意义(P<0.05,P<0.01)。结论辛开苦降法治疗RE模型大鼠的作用机制可能与其下调iNOS的蛋白表达相关。

Objective To explore the impacts of xinkaikujiang therapy on iNOS protein expression in the lower esophageal tissue of the rat model of reflux esophagitis( RE). Methods Ninety- eight adult male wistar rats were randomized into a normal group( 10 rats) and a model group( 88 rats). The modified partial cardiomyotomy plus external pylorus semi- ligation was adopted in modeling. After modeling,the dead and weak rats were dropped out. Sixty rats of good recovery were randomized into a model group,the low-dose,middle- dose and high- dose Chinese medicine groups,a mixture group of Chinese and western medicine and a western medicine group,10 rats in each one. No any treatment was applied in the normal group.The lavage was given in each group. The physical saline 0. 9% was used in the model group,the modified banxiaxiexin decoction with xinkaikujiang therapy was used with different concentrations in the low- dose,middle- dose and high- dose Chinese medicine groups. The mixture liquor with domperidone,hydrotalcite and rabeprazole sodium was used in the western medicine group. The middle- dose Chinese medicine and the mixture liquor as the western medicine group were used in the mixture group of Chinese and western medicine. According to the treatment time,each group was subdivided into a 7 d subgroup and a 14 d subgroup,5rats in each one. The tissues were collected on the 7thand the 14 thdays separately. IHC and western- blot methods were adopted to determine iNOS expression in the lower esophageal mucosa in each group. Results As compared with the normal group,iNOS expression was increased in the model group( P < 0. 05). In 7 days of treatment,as compared with the model groups,iNOS expression was not different significantly in each group( P > 0. 05). In 14 days of treatment,as compared with the model group,iN OS expression was reduced to different extents in the high- dose Chinese medicine group,the mixture group of Chinese and western medicine and the western medicine group,indicating the significant differences( P < 0. 05,P < 0. 01). Conclusion Xinkaikujiang therapy treats RE in model rats probably relevant with the down- regulation of iN OS protein expression.