清中愈疡汤对脾胃湿热型胃溃疡作用的实验研究

Experimental Study on the Effect of Qingzhong Yuyang Decoction on Gastric Ulcer of Spleen and Stomach Damp-heat Type

目的:探讨清中愈疡汤对脾胃湿热型胃溃疡的治疗作用,为临床用药提供实验依据。方法:将60只Wistar大鼠随机分为6组,正常对照组、模型对照组、清中愈疡汤高剂量组、清中愈疡汤中剂量组、清中愈疡汤低剂量组、奥美拉唑组,每组10只。正常对照组正常饲养,其余各组均制作为实验性脾胃湿热型胃溃疡大鼠模型。造模成功予以持续治疗30 d后,处死大鼠取其血样和胃黏膜组织。测量计算大鼠溃疡面积及溃疡抑制率;将胃黏膜组织制成病理切片行苏木素伊红(HE)染色,在显微镜下观察各组大鼠胃黏膜组织的病理学变化,并检测大鼠血清中肿瘤坏死因子(TNF-α)、白介素-8(IL-8)、胃动素(MTL)、前列腺素E2(PGE2)等水平,比较干预前后各组相关指标变化情况。通过Western blot法检测大鼠胃黏膜组织中核转录因子κB(NF-κB)通路相关蛋白水平,包括磷酸化核因子κB亚基65(p-NF-κB p65)、磷酸化核因子κB抑制蛋白(p-IκBα)。结果:与正常对照组比较,模型对照组胃黏膜发生明显病理变化,黏膜上皮细胞明显缺损,可见黏膜组织充血及水肿现象,并存在明显炎性细胞浸润。干预后,各组溃疡面积、溃疡抑制率、TNF-α、IL-8、MTL及PGE2水平、NF-κB p65、p-NF-κB p65、IκBα、p-IκBα蛋白表达水平比较,差异均有统计学意义(P<0.05);干预后,与正常对照组比较,模型对照组溃疡面积增加,NF-κB p65、IκBα蛋白表达水平均降低,p-NF-κB p65、p-IκBα蛋白表达水平均升高(P<0.05);干预后,与模型对照组比较,清中愈疡汤高、中、低剂量组及奥美拉唑组溃疡面积均明显减少,TNF-α、IL-8水平均下降,MTL、PGE2水平均升高,(P<0.05);与奥美拉唑组比较,清中愈疡汤高、中剂量组溃疡面积均明显减少,溃疡抑制率均明显增加(P<0.05);与清中愈疡汤低剂量组比较,清中愈疡汤高、中剂量组溃疡面积均明显减少,溃疡抑制率均明显增加(P<0.05)。与清中愈疡汤中剂量组比较,清中愈疡汤高剂量组明显减少,溃疡抑制率明显增加(P<0.05)。与模型对照组比较,清中愈疡汤高、中、低剂量组均出现不同程度的NF-κB p65、IκBα蛋白表达水平升高、p-NF-κB p65下降;清中愈疡汤高剂量组p-IκBα蛋白表达水平低于模型对照组(P<0.05)。结论:清中愈疡汤可通过抑制炎症因子释放以及抑制NF-κB信号通路相关蛋白表达,以减少溃疡面积,并促进胃黏膜内源性物质分泌以及胃活动功能,从而达到对脾胃湿热型胃溃疡的治疗作用,促进溃疡愈合。

Objective:To explore the therapeutic effect of Qingzhong Yuyang Decoction on gastric ulcer of spleen-stomach damp-heat type,and to provide experimental basis for clinical medication.Method:A total of 60 Wistar rats were randomly divided into 6 groups:normal control group,model control group,high-dose Qingzhong Yuyang Decoction high-dose group,Qingzhong Yuyang Decoction medium-dose group,Qingzhong Yuyang Decoction low-dose group,Omeprazde group,10 animals in each group.The normal control group was fed normally,and the other groups were used as experimental gastric ulcer rat models of spleen-stomach dampheat type.After successful modeling and continuous treatment for 30 days,the rats were sacrificed to obtain blood samples and gastric mucosal tissue.The ulcer area and the ulcer inhibition rate of the rats were measured and calculated;the gastric mucosa tissue was made into pathological sections and stained with hematoxylin and eosin(HE),the pathological changes of the gastric mucosa tissue of the rats in each group were observed under a microscope,and the serum levels of the rats were detected.The levels of tumor necrosis factor(TNF-α),interleukin-8(IL-8),motilin(MTL),and prostaglandin E2(PGE2) were compared,and the changes of related indicators in each group before and after the intervention were compared.Western blot was used to detect the levels of nuclear factor κB(NF-κB) pathway-related proteins in rat gastric mucosa,including phosphorylated nuclear factor κB subunit 65(p-NF-κB p65),phosphorylated nuclear factor κB inhibitory protein( p-IκBα).Result:Compared with normal control group,gastric mucosa in model control group showed obvious pathological changes,with obvious defect of epithelial cells,hyperemia and edema of mucosal tissue,and obvious inflammatory cell infiltration.After intervention,there were significant differences in ulcer area,ulcer inhibition rate,TNF-α,IL-8,MTL and PGE2 levels,and protein expression levels of NF-κB p65,P-NF-κB p65,IκBα and P-I κBαamong all groups(P<0.05).After intervention,compared with normal control group,ulcer area increased,NF-κB p65 and IκBα protein expression decreased,and P-NF-κB p65 and P-I κBα protein expression increased in model control group(P<0.05).After intervention,compared with model control group,ulcer area in Qingzhong Yuyang Decoction high-dose,medium-dose and low-dose groups and Omeprazole groups were significantly reduced,TNF-αand IL-8 levels were decreased,MTL and PGE2 levels were increased(P<0.05).Compared with omeprazole group,ulcer area in Qingzhong Yuyang Decoction high-dose and medium-dose groups was significantly reduced,and ulcer inhibition rate was significantly increased(P<0.05).Compared with the Qingzhong Yuyang Decoction low-dose group,the ulcer area in the Qingzhong Yuyang Decoction high-dose and medium-dose groups were significantly reduced,and the ulcer inhibition rate were significantly increased(P<0.05).Compared with Qingzhong Yuyang Decoction medium-dose group,the ulcer inhibition rate in Qingzhong Yuyang Decoction high-dose group was significantly increased(P<0.05).Compared with model control group,protein expression levels of NF-κB p65 and IκBα were increased and p-NF-κB p65 was decreased in Qingzhong Yuyang Decoction high-dose,medium-dose and lowdose groups.The protein expression level of P-I κBα in Qingzhong Yuyang Decoction high-dose group was lower than that in model control group(P<0.05).Conclusion:Qingzhong Yuyang Decoction can reduce the ulcer area by inhibiting the release of inflammatory factors and inhibiting the expression of NF-κB signaling pathway-related proteins,and promote the secretion of endogenous substances in gastric mucosa and gastric motility function,so as to achieve the therapeutic effect on gastric ulcer of spleen-stomach damp-heat type,and promote ulcer healing.