六君子汤治疗慢性萎缩性胃炎大鼠疗效和机制研究

Effect and Mechanism of Liujunzi Decoction on Chronic Atrophic Gastritis in Rats

目的探讨六君子汤治疗慢性萎缩性胃炎(CAG)大鼠的疗效和作用机制。方法将22只SPF级Wistar大鼠根据体质量随机分为正常组5只和造模组17只。正常组不予造模,造模组予以0.1%氨水溶液及20 mmol/L脱氧胆酸钠溶液交替饮用联合饥饱失常法造模16周后,光镜下观察随机2只造模组大鼠胃黏膜组织形态改变以判定造模成功。造模组造模成功后根据体质量随机分为模型组、维酶素组、六君子汤组,每组各5只。六君子汤组按体质量10 mL/(kg·d)灌胃0.405 g/mL六君子汤水煎液,维酶素组按体质量10 mL/(kg·d)灌胃24 mg/mL维酶素混悬液,正常组和模型组灌胃等剂量灭菌饮用水,连续干预4周。干预后观察4组大鼠一般情况和测量体质量,光镜下观察4组大鼠胃黏膜组织形态,ELISA法检测4组大鼠血清肿瘤坏死因子-α(TNF-α)含量,qPCR检测4组大鼠胃黏膜组织增殖细胞核抗原(PCNA)、血管内皮生长因子(VEGF)、嗜银蛋白(Ag-NOR)和促细胞凋亡基因(Bax)mRNA相对表达水平。结果与正常组比较,模型组活动状况、毛发色泽、大便情况等均无改善,体质量较正常组降低(P<0.01),胃黏膜腺体排列紊乱、萎缩,炎性细胞浸润,血清TNF-α含量及胃黏膜组织PCNA、VEGF、Ag-NOR、Bax mRNA相对表达水平均升高(P均<0.05或0.01);与模型组比较,维酶素组及六君子汤组活动状况、毛发色泽、大便情况等均改善,体质量均升高(P均<0.05),胃黏膜排列较紧密,腺体萎缩有所改善,炎症细胞浸润有所减少,血清TNF-α含量及胃黏膜组织PCNA、VEGF、Ag-NOR、Bax mRNA相对表达水平均降低(P均<0.05或0.01)。结论六君子汤可以改善CAG大鼠胃黏膜病理形态,其机制可能与下调炎症因子TNF-α,抑制增殖因子PCNA、Ag-NOR、VEGF和促凋亡基因Bax的表达有关。

Objective:To explore the therapeutic effect and mechanism of Liujunzi decoction on chronic atrophic gastritis(CAG).Methods:A total of 22 SPF grade Wistar rats were randomly divided into normal group of 5 and model group of 17.The normal group did not go through modeling,but the model group was given 0.1%ammonia water solution and 20 mmol/L sodium deoxycholate solution for alternating drinking,combined with the method of hunger and satiety disorders.After 16 weeks of modeling,the morphological changes in the gastric antrum tissue of two randomly selected model group rats were observed under light microscope to determine the success of modeling.After successful modeling,the modeling group was randomly divided into model group,vitacoenzyme group,and Liujunzi decoction group,with 5 rats in each group.The Liujunzi decoction group was given 10 mL/(kg·d)by gavage of 0.405 g/mL Liujunzi decoction,the vitacoenzyme group was given 10 mL/(kg·d)by gavage of 24 mg/mL vitacoenzyme suspension,the normal group and the model group were given equal volume of sterile drinking water by gavage,and the intervention lasted for 4 weeks.After intervention,the general condition and body weight of rats were observed,the histological morphology of gastric mucosa was observed under light microscope,ELISA was used to detect TNF-αin serum of rats in each group,the relative expression levels of proliferating cell nuclear antigen(PCNA),vascular endothelial growth factor(VEGF),argyrophilic protein(Ag-NOR)and pro apoptotic gene(Bax)mRNA in rat gastric mucosa were detected by real-time fluorescent quantitative PCR.Results:Compared with the normal group,the activity,hair color and stool of the model group did not improve,and the body mass was lower than that of the normal group(P<0.01),the glands of gastric mucosa were disordered and atrophied,and inflammatory cells infiltrated,the congtent of serum TNF-αand the relative expression levels of PCNA,VEGF,Ag NOR and Bax mRNA in gastric mucosa were higher than those in the normal group(P<0.05 or 0.01).Compared with the model group,the activity,hair color,stool condition and body weight of the vitacoenzyme group and Liujunzi decoction group improved,and the body weight increased(P<0.05),gastric mucosa was tightly packed,glandular atrophy improved,and inflammatory cell infiltration is reduced,the content of serum TNF-αand the relative expression levels of PCNA,VEGF,Ag-NOR and Bax mRNA in gastric mucosa were lower than those in the model group(P<0.05 or 0.01).Conclusion:Liujunzi decotion can improve the pathological morphology of gastric mucosa in CAG rats,and its mechanism may be related to the down-regulation of inflammatory factor,inhibition of the expression of proliferative factors PCNA,Ag-NOR,VEGF,and Bax.