黄芪建中汤对脾胃虚寒型胃溃疡大鼠作用机制研究

目的研究黄芪建中汤对脾胃虚寒型胃溃疡(GU)模型大鼠胃组织炎性因子以及缺氧诱导因子-1α(HIF-1α)、环氧化酶-2(COX-2)、诱导型一氧化氮合酶(iNOS)和内皮型一氧化氮合酶(eNOS)蛋白表达水平的影响。方法 Wistar大鼠用综合造模法复建脾胃虚寒型GU模型。按随机数字表法将模型大鼠分为5组:模型组、阳性对照组(0.14 g·kg-1·d-1安胃疡灌胃)和高、中、低3个剂量实验组(16,8,4 g·kg-1·d-1黄芪建中汤灌胃),每组10只;另选正常10只大鼠作为空白组。空白组和模型组大鼠给予10 g·kg-1·d-1蒸馏水灌胃,持续给药21 d。用蛋白质印迹法检测胃组织HIF-1α和COX-2蛋白表达水平(灰度值),免疫组化法检测胃组织iNOS和eNOS蛋白表达水平(光密度值)。结果空白组、模型组、阳性对照组和高、中、低3个剂量实验组的HIF-1α蛋白表达量分别为1.30±0.14,2.18±0.29,1.71±0.14,1.45±0.14,1.79±0.15和1.97±0.22;此6组的COX-2蛋白表达量分别为1.38±0.19,3.33±0.21,2.52±0.15,2.27±0.19,2.62±0.04和3.36±0.54;此6组的iNOS蛋白表达量分别为16.29±1.55,22.46±1.64,20.27±1.74,17.74±1.11,19.22±1.70和21.09±2.21;此6组的eNOS蛋白表达量分别为22.43±1.57,16.69±1.46,18.82±2.71,19.55±2.07,19.18±1.77和18.01±2.09。上述指标:模型组与空白组比较,组间差异均有统计学意义(均P<0.05)。与模型组比较,阳性对照组和高、中2个剂量实验组均下调了HIF-1α、COX-2和iNOS蛋白表达水平,上调了eNOS蛋白表达水平,差异均有统计学意义(均P<0.05)。结论黄芪建中汤改善脾胃虚寒型GU模型大鼠的作用机制可能与有效下调HIF-1α、COX-2、iNOS蛋白表达水平和上调eNOS蛋白表达水平有关,进而促进溃疡病灶黏膜损伤的修复。

Effect and mechanism of Huangqi Jianzhong Decoction on gastric ulcer of spleen stomach deficiency cold type in rats

Objective To investigate the effect of Huangqi Jianzhong Decoction on the expression of inflammatory factors and hypoxia inducible factor-1 α(HIF-1 α), cyclooxygenase-2(COX-2), inducible nitric oxide synthase(iNOS), endothelial nitric oxide synthase(eNOS) in gastric tissue of rats with gastric ulcer(GU).Methods Wistar rats were used to establish GU model of deficiency cold of spleen and stomach.The model rats were divided into 5 groups according to the random number table method: model group, positive control group(0.14 g·kg-1·d-1) and high, medium and low dose experimental groups (16,8,4 g·kg-1·d-1 Huangqi Jianzhong Decoction) with 10 rats in each group;another 10 normal rats were selected as the blank group.The rats in the blank group and model group were given 10 m L·kg-1·d-1 distilled water by gavage for 21 d.The expression levels(gray value) of HIF-1 α and COX-2 in gastric tissue were detected by Western blot method,and the expression levels(OD value) of i NOS and e NOS in gastric tissue were detected by immunohistochemistry.Results The expression of HIF-1 α protein in normal group,model group,positive control group and high,medium,low dose experimental groups were 1.30 ± 0.14,2.18 ± 0.29,1.71 ± 0.14,1.45 ± 0.14,1.79 ± 0.15,1.97 ± 0.22,respectively;the expression of COX-2 protein in the six groups were 1.38 ± 0.19,3.33 ± 0.21,2.52 ± 0.15,2.27 ± 0.19,2.62 ± 0.04,3.36 ± 0.54,respectively;the expression of i NOS in the six groups were 16.29 ± 1.55,22.46 ± 1.64,20.27 ± 1.74,17.74 ± 1.11,19.22 ± 1.70,21.09 ± 2.21,respectively;the expression of e NOS in the six groups were 22.43 ± 1.57,16.69 ± 1.46,18.82 ± 2.71,19.55 ± 2.07,19.18 ± 1.77,18.01 ± 2.09,respectively.The above indexes: compared with the blank group,the expression level of HIF-1α,COX-2,iNOS protein of the model group was significantly increased,and the expression level of e NOS protein was significantly reduced,and the difference between the groups was statistically significant(all P < 0.05).Compared with the model group,the expression level of HIF-1α,COX-2,iNOS protein was down regulated and the expression level of e NOS protein was up regulated in positive control group and high,medium dose experimental groups,the difference was statistically significant(all P < 0.05).Conclusion The specific mechanism of Huangqi Jianzhong Decoction in the treatment of GU model of spleen and stomach deficiency cold may be that it can effectively down regulate the expression level of HIF-1α,COX-2,iNOS protein,up regulate the expression level of eNOS protein,and then promote the repair of ulcerative lesions.