戊己丸对吲哚美辛诱导大鼠胃溃疡的保护作用研究

Protective Effect of Wuji Pill(戊己丸)on Indomethacin-Induced Gastric Ulcer in Rats

目的 研究戊己丸对吲哚美辛诱导的大鼠胃溃疡的保护作用,并初步探索其可能的作用机制。方法 36只SD大鼠随机分为正常组、模型组、阳性对照组(奥美拉唑,20 mg/kg)、戊己丸低、中、高(2.52、5.04、10.08 g/kg生药量)剂量组,每组6只。各组大鼠灌胃相应药物或0.5%羧甲基纤维素钠(Carboxymethyl Cellulose, CMC-Na)水溶液,1次/d,连续7 d,于末次给药1 h后,除正常组外,其余各组均给予吲哚美辛(60 mg/kg)灌胃造模;6 h后处死大鼠,收集血清和胃组织。通过改良Guth法计算胃溃疡指数和溃疡抑制率,染色观察比较各组大鼠胃组织病理学变化;使用试剂盒测定胃组织中丙二醛(Malonaldehyde, MDA)、超氧化物歧化酶(Superoxide Dismutase, SOD)和环氧合酶1(Cyclooxygenase-1,COX-1),血清中一氧化氮(Nitric Oxide, NO)、肿瘤坏死因子(Tumor Necrosis Factor-α,TNF-α)、白细胞介素-6(Interleukin-6,IL-6)和白细胞介素-10(Interleukin-10,IL-10)的含量;TUNEL染色法比较各组大鼠胃组织细胞凋亡情况;免疫组化法测定胃组织中环氧合酶2(Cyclooxygenase-2,COX-2)、NF-κB(p65)蛋白的表达水平。结果 与正常组相比,模型组大鼠胃组织黏膜层、黏膜下层、肌层及浆膜部分结构受损,胃溃疡指数和胃组织细胞凋亡百分比显著升高(P<0.01);MDA、TNF-α、IL-6水平显著升高(P<0.01),COX-1、NO、IL-10和SOD水平显著降低(P<0.01);COX-2和NF-κB(p65)蛋白表达显著升高(P<0.01);与模型组相比,各给药组均能改善胃组织损伤状况;戊己丸中、高剂量组和奥美拉唑组均能显著降低(P<0.01)胃溃疡指数和升高溃疡抑制率,升高COX-1、NO、IL-10水平,降低MDA、COX-2、TNF-α、IL-6和NF-κB(p65)水平,各给药组均明显降低大鼠胃组织细胞凋亡百分比,其中戊己丸中剂量组表现出良好的作用。结论 戊己丸对吲哚美辛诱导的大鼠胃溃疡有明显的保护作用,其作用机制可能与改善氧化应激抑制、炎症反应和减少细胞凋亡有关。

Objective This study aimed to investigate the protective effect of Wuji Pill(戊己丸)on indomethacin-induced gastric ulcer in rats and explore its possible mechanism.Methods A total of 36 SD rats were randomly divided into normal group,model group,positive control group(omeprazole,20 mg/kg),Wuji Pill low,middle and high dose groups,with 6 rats in each group.The rats in each group were given corresponding drugs or 0.5%carboxymethyl cellulose(CMC-Na)water solution once a day for 7 days.After the last administration for 1 hour,except the normal group,indomethacin(60 mg/kg)was given to all oth-er groups to make models.After 6 hours,the rats were killed and the blood and stomach tissues were collected.The gastric ulcer index and ulcer inhibition rate were calculated by modified Guth method,and the pathological changes of gastric tissue of rats in each group were observed and compared by HE and PAS staining.The contents of malondialdehyde(MDA),superoxide dis-mutase(SOD)and cyclooxygenase-1(COX-1)in stomach tissue,nitric oxide(NO),tumor necrosis factor-α(TNF-α),in-terleukin-6(IL-6)and interleukin-10(IL-10)in serum were determined by using the corresponding kit.TUNEL staining was used to compare the apoptosis of gastric tissue cells in each group.The expression levels of cyclooxygenase-2(COX-2)and nuclear factor kappa-Bp65[NF-κB(p65)]protein in gastric tissue were detected by immunohistochemistry.Re-sults Compared with those of the normal group,some structures of gastric mucosa,submucosa,muscular layer and serosa in the model group were damaged,and the gastric ulcer index and apoptosis percentage of gastric tissue cells were significantly increased(P<0.01).The levels of MDA,TNF-αand IL-6 increased significantly(P<0.01),while the levels of COX-1,NO,IL-10 and SOD decreased significantly(P<0.01).The expressions of COX-2 and NF-κB(p65)protein increased significantly(P<0.01).Compared with the model group,all groups can improve the gastric tissue injury.The middle and high dose groups of Wuji Pill and omeprazole group can significantly reducethe gastric ulcer index(P<0.01),increase the ulcer inhibition rate,in-crease the expression levels of COX-1,NO and IL-10,and reduce the levels of MDA,COX-2,TNF-α,IL-6 and NF-κB(p65).All groups of Wuji Pill can significantly reduce the percentage of apoptosis in gastric tissue in rats,and the middle dose group of Wuji Pill has a good effect.Conclusion Wuji Pill has obvious protective effect on indomethacin-induced gastric ulcer in rats,and its mechanism may be related to improving oxidative stress,inflammation and reducing apoptosis.