基于抗氧化作用观察胃血止糊剂对应激性溃疡模型大鼠的保护作用和机制

Study on protection and mechanism of gastric blood check paste on stress ulcer in model rats:based on antioxidation

目的观察胃血止糊剂对大鼠应激性溃疡(SU)的保护作用和机制。方法将72只SPF级SD雄性大鼠随机分为空白组、模型组、奥美拉唑组及胃血止糊剂高、中、低剂量组共6组,每组12只,奥美拉唑组予奥美拉唑5 mg/(kg·d)灌胃,胃血止糊剂高、中、低剂量组分别予胃血止糊剂4、2、1 g/(kg·d)灌胃,空白组和模型组予等容积的溶媒灌胃,每日1次,共灌胃14 d。灌胃后除空白组外,其余组采用“水浸束缚应激”制备SU模型。采用激光多普勒测定各组大鼠建模后胃黏膜血流量,制作苏木素-伊红染色切片观察各组胃黏膜组织病理形态学变化;采用酶联免疫吸附法检测各组大鼠血清及胃黏膜组织中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)活性及丙二醛(MDA)含量。结果与空白组比较,模型组、奥美拉唑组和胃血止糊剂高、中、低剂量组GMBF均降低(P<0.05),UI均升高(P<0.05);与模型组比较,奥美拉唑组和胃血止糊剂高、中、低剂量组GMBF均升高(P<0.05),奥美拉唑组和胃血止糊剂高、中剂量组UI均降低(P<0.05);与奥美拉唑组、胃血止糊剂高剂量组比较,胃血止糊剂中、低剂量组GMBF均降低(P<0.05),与奥美拉唑组比较,胃血止糊剂中、低剂量组UI均升高(P<0.05),胃血止糊剂高剂量组GMBF、UI与奥美拉唑组比较差异无统计学意义(P>0.05);与胃血止糊剂中剂量组比较,胃血止糊剂低剂量组GMBF显著降低(P<0.05),UI无差异(P>0.05)。空白组大鼠胃黏膜组织未见损伤性的变化,胃黏膜结构完整,腺体排列整齐;模型组大鼠胃黏膜损伤明显,上皮细胞广泛坏死脱落,腺体结构遭到破坏,形成多个大小、深浅不等的糜烂面。奥美拉唑组和胃血止糊剂各剂量组胃黏膜损伤较模型组明显减轻。与空白组比较,模型组、奥美拉唑组及胃血止糊剂高、中、低剂量组大鼠血清SOD、GSH-Px、CAT活性均降低(P<0.05),MDA含量均升高(P<0.05);与模型组比较,胃血止糊剂高、中、低剂量组大鼠血清SOD、GSH-Px活性均增加(P<0.05),MDA含量均降低(P<0.05),胃血止糊剂高、中剂量组大鼠血清CAT活性均增加(P<0.05),奥美拉唑组各指标无变化(P>0.05);与奥美拉唑组比较,胃血止糊剂高、中、低剂量组大鼠血清SOD活性均增加(P<0.05),MDA含量均降低(P<0.05),胃血止糊剂高剂量组大鼠血清CAT活性增加(P<0.05);与胃血止糊剂高剂量组比较,胃血止糊剂中、低剂量组大鼠血清SOD活性均降低(P<0.05)。与空白组比较,模型组、奥美拉唑组及胃血止糊剂高、中、低剂量组大鼠胃黏膜组织SOD、GSP-Px、CAT活性均降低(P<0.05),MDA含量均升高(P<0.05);与模型组比较,胃血止糊剂高、中、低剂量组大鼠胃黏膜组织SOD、GPS-Px活性均增加(P<0.05),MDA含量均降低(P<0.05),胃血止糊剂高、中剂量组大鼠胃黏膜组织CAT活性均增加(P<0.05);与奥美拉唑组比较,胃血止糊剂高、中、低剂量组大鼠胃黏膜组织SOD、GSP-Px活性均增加(P<0.05),MDA含量均降低(P<0.05),胃血止糊剂高、中剂量组大鼠胃黏膜组织CAT活性均增加(P<0.05);与胃血止糊剂高剂量组比较,胃血止糊剂中、低剂量组大鼠胃黏膜组织SOD、GSH-Px活性均降低(P<0.05),胃血止糊剂低剂量组大鼠胃黏膜组织MDA含量升高(P<0.05);与胃血止糊剂中剂量组比较,胃血止糊剂低剂量组大鼠胃黏膜组织SOD、CAT活性均降低(P<0.05),MDA含量升高(P<0.05)。结论胃血止糊剂能改善SU模型大鼠胃黏膜损伤,具有明显的抗氧化作用,且其抗氧化作用可能是主要通过增加SOD和GSH-Px的活性来实现的,对CAT活性的调节可能也是其发挥抗氧化作用的原因之一。

Objective To observe the protection and mechanism of gastric blood check paste on stress ulcer(SU)in rats.Methods Totally 72 SPF-grade SD male rats randomly assigned into the blank group,the model group,the omeprazole group,the high-dose,medium-dose and low-dose gastric blood check paste groups,with 12 per group.The omeprazole group was given omeprazole(5 mg/[kgod])by gavage;the high-dose,medium-dose and low-dose gastric blood check paste groups were given gastric blood check paste(4,2,and 1 g/[kgod],respectively)by gavage;the blank group and the model group received equal volume of dissolvent;the treatment continued for 14 days(qd).The SU model was established by water-immersion stress in the other groups apart from the blank group.The gastric mucosal blood flow was measured with laser Doppler flowmeter;the pathomorphism of gastric mucosa was observed using hematoxylin-eosin-stained sections;the superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),Catalase(CAT),malonaldehyde(MDA)of serum and gastric mucosa were determined by enzyme-linked immunosorbent assay(ELISA).Results In comparison of the blank group and the other groups,the other groups displayed decreased gastric mucosal blood flow(GMBF),and increased ulcer index(UI)(all P<0.05);compared with the model group,GMBF in the omeprazole group,the high-,medium-and low-dose gastric blood check paste groups was increased(P<0.05),UI in the omeprazole group,the high-,medium-dose gastric blood check paste groups was decreased(P<0.05);lower GMBF were detected in the medium-and low-dose gastric blood check paste groups than those in the the omeprazole group,the high-dose gastric blood check paste group,and higher UI were detected in the medium-and low-dose gastric blood check paste groups than those in the the omeprazole group(all P<0.05);none of the GMBF and UI differed significantly between the high-dose gastric blood check paste group and the omeprazole group(P>0.05);GMBF in the low-dose gastric blood check paste group was significantly decreased than that in the medium-dose gastric blood check paste group(P<0.05),and no difference in UI was found(P>0.05).The gastric mucosa in the blank group showed no damage,with a complete gastric mucosa structure and neat gland arrangement;the gastric mucosa in the model group was significantly damaged,with extensive necrotic and exfoliated epithelial cells,and damaged gland structure,and multiple erosion surfaces of varying sizes and depths were formed.The gastric mucosal injury in the omeprazole group and the gastric blood check paste groups was significantly reduced compared with the model group.Decreased serum SOD,GSH-Px and CAT,and increased MDA were measured in the other groups than those in the blank group(all P<0.05).In comparison with the model group,elevated serum SOD and GSH-Px,and reduced MDA in the high-,medium-and low-dose gastric blood check paste groups were found(P<0.05),elevated serum CAT in the high-,medium-dose gastric blood check paste groups was detected(P<0.05),no change in each indicator in the omeprazole group was found(P>0.05).Compared with the omeprazole group,serum SOD in the high-,medium-and low-dose gastric blood check paste groups was increased(P<0.05),while MDA was decreased(P<0.05),serum CAT in the high-dose gastric blood check paste group was increased(P<0.05);serum SOD in the medium-and low-dose gastric blood check paste groups was decreased than that in the high-dose gastric blood check paste group(P<0.05).Decreased SOD,GSH-Px and CAT of gastric mucosa,and increased MDA were measured in the other groups than those in the blank group(all P<0.05).Compared with the model group,higher SOD and GSH-Px of gastric mucosa,and lower MDA in the high-,medium-and low-dose gastric blood check paste groups were detected(P<0.05),higher CAT of gastric mucosa in the high-,medium-dose gastric blood check paste groups was detected(P<0.05).In comparison with the omeprazole group,SOD and GSP-Px of gastric mucosa in the high-,medium-and low-dose gastric blood check paste groups were increased(P<0.05),while MDA was decreased(P<0.05),CAT of gastric mucosa in the high-,medium-dose gastric blood check paste group was increased(P<0.05).Lower SOD and GSH-Px of gastric mucosa in the medium-and low-dose gastric blood check paste groups,and higher MDA of gastric mucosa in the low-dose gastric blood check paste group were detected than those in the high-dose gastric blood check paste group(all P<0.05).SOD,CAT of gastric mucosa in the low-dose gastric blood check paste group were decreased compared with the medium-dose gastric blood check paste group(P<0.05),while MDA was increased(P<0.05).Conclusion The antioxidant effect of gastric blood check paste is more obvious,which may be mainly achieved by increasing the activities of SOD and GSH-Px.The regulation of CAT by gastric blood check paste may also be the reason for its antioxidation.