摘要
目的:探讨柴芍六君汤对肝郁脾虚型慢性萎缩性胃炎(CAG)大鼠刺猬(Hedgehog)信号通路的影响。方法:随机将Wistar大鼠分为正常组和造模组,造模组采用复合造模法进行构建肝郁脾虚型CAG大鼠模型,造模成功后,分为模型组,维酶素组,柴芍六君汤组,GDC-0449(阻滞剂)组,柴芍六君汤+GDC-0449组;正常组和模型组给予生理盐水灌胃,维酶素组和柴芍六君汤组分别给予240 mg·kg^(-1)·d^(-1),5.1 g·kg^(-1)·d^(-1)灌胃,GDC-0449组腹腔注射50 mg·kg^(-1)·d^(-1),柴芍六君汤+GDC-0449给予腹腔注射50 mg·kg^(-1)·d^(-1)和灌胃柴芍六君汤5.1 g·kg^(-1)·d^(-1),持续4周。苏木素-伊红(HE)染色观察大鼠胃黏膜病理形态,实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测胃黏膜组织音猬因子(Shh),12次跨膜蛋白受体Patched1(Ptch1),胶质瘤相关肿瘤基因同源物1(Gli1)m RNA和蛋白表达,酶联免疫吸附测定法(ELISA)检测血清白细胞介素-1β(IL-1β),肿瘤坏死因子-α(TNF-α)含量。结果:与正常组比较,模型组大鼠腺细胞减少,腺体萎缩,腺腔体积增大,可见浆细胞浸润,伴有肠上皮化生,胃黏膜组织Shh,Ptch1,Gli1 m RNA和蛋白表达显著降低(P<0.01),血清中IL-1β,TNF-α含量显著上升(P<0.01)。与模型组比较,维酶素组、柴芍六君汤组细胞排列较为整齐,腺体萎缩改善,未见明显炎性浸润,GDC-0499组和柴芍六君汤+GDC-0449组并无太多改善;维酶素组、柴芍六君汤组胃黏膜组织Shh,Ptch1,Gli1 m RNA和蛋白表达水平显著上升(P<0.01),维酶素组血清中IL-1β含量差异无统计学意义,TNF-α含量显著降低(P<0.01),柴芍六君汤组血清中IL-1β,TNF-α含量明显降低(P<0.05,P<0.01);GDC-0449组和柴芍六君汤+GDC-0449组,胃黏膜组织Shh,Ptch1,Gli1 m RNA和蛋白表达、血清中IL-1β,TNF-α含量差异无统计学意义。结论:柴芍六君汤能有效改善肝郁脾虚型大鼠胃黏膜病理学状态,其机制可能是通过激活Hedgehog信号通路,从而降低IL-1β,TNF-α含量有关。
Objective:To explore the effect of Chaishao Liujuntang on Hedgehog signaling pathway in rats with chronic atrophic gastritis(CAG)of liver depression and spleen deficiency.Method:Wistar rats were randomized into normal group and modeling group.CAG with the liver depression and spleen deficiency syndrome was induced in rats in the modeling group with a compound method.After modeling,they were classified into the model group,vitacoenzyme group,Chaishao Liujuntang group,GDC-0449(blocker)group,and Chaishao Liujuntang+GDC-0449 group.Normal group and model group were given(ig)normal saline.Vitacoenzyme and Chaishao Liujuntang group received(ig)corresponding drugs at 240 mg·kg^(-1)·d^(-1)and5.1 g·kg^(-1)·d^(-1),respectively,and GDC-0449 group was treated(ip)with GDC-0449 at 50 mg·kg^(-1)·d^(-1).For the Chaishao Liujuntang+GDC-0449 group,rats received GDC-0449(ip)at 50 mg·kg^(-1)·d^(-1)and Chaishao Liujuntang(ig)at 5.1 g·kg^(-1)·d^(-1).The administration lasted 4 weeks.The pathological morphology of rat gastric mucosa was observed based on hematoxylin-eosin(HE)staining.m RNA and protein expression of sonic hedgehog(Shh),12th transmembrane receptor Patched1(Ptch1),and glioma-associated oncogene homolog 1(Gli1)in gastric mucosa tissues was detected by real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)and Western blot.Content of serum interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)was determined by enzyme-linked immunosorbent assay(ELISA).Result:Compared with normal group,the model group demonstrated decrease in gland cells,glandular atrophy,large lumen volume,plasma cell infiltration,intestinal metaplasia,decrease in the m RNA and protein expression of Shh,Ptch1,and Gli1 in gastric mucosa(P<0.01),and rise of serum IL-1βand TNF-αcontent(P<0.01).Compared with model group,vitacoenzyme group and Chaishao Liujuntang group showed ordered cells,alleviation of gland atrophy,and no obvious inflammatory infiltration,and GDC-0499 group and Chaishao Liujuntang+GDC-0449 showed no significant improvement.Significant rise in the m RNA and protein expression of Shh,Ptch1,and Gli1 in gastric mucosa tissues of vitacoenzyme group and Chaishao Liujuntang group(P<0.01),no significant difference in serum IL-1βcontent and significant decrease in TNF-αcontent in vitacoenzyme group(P<0.01),significant reduction in content of serum IL-1βand TNF-αin Chaishao Liujuntang group(P<0.05,P<0.01)were observed compared with those in the model group.The m RNA and protein expression of Shh,Ptch1,and Gli1 in gastric mucosa and the content of serum IL-1βand TNF-αwere insignificantly different between the GDC-0449 group and Chaishao Liujuntang+GDC-0449 group.Conclusion:Chaishao Liujuntang can effectively improve the pathological state of gastric mucosa in CAG rats with liver depression and spleen deficiency,which may be related to the activation of Hedgehog signaling pathway and the decrease of IL-1βand TNF-αcontent.
作者
涂文玲
黄妙安
洪银洁
洪诗晓
陈梅妹
甘慧娟
TU Wen-ling;HUANG Miao-an;HONG Yin-jie;HONG Shi-xiao;CHEN Mei-mei;GAN Hui-juan(College of Traditional Chinese Medicine(TCM),Fujian University of TCM,Fuzhou 350122,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第4期1-8,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81873237)
福建省大学生创新创业训练计划项目(202010393025)
福建中医药大学校管课题(X2020012)
福建省自然科学基金项目(2021J0113)。
