BACKGROUND Gastric intestinal metaplasia(IM)is a precancerous lesion that is associated with an elevated risk of gastric carcinogenesis.Weiwei Decoction(WWD)is a promising traditional Chinese herbal formula widely emp...BACKGROUND Gastric intestinal metaplasia(IM)is a precancerous lesion that is associated with an elevated risk of gastric carcinogenesis.Weiwei Decoction(WWD)is a promising traditional Chinese herbal formula widely employed in clinical for treating IM.Previous studies suggested the potential involvement of the olfactomedin 4(OLFM4)/nucleotide-binding oligomerization domain 1(NOD1)/caudal-type homeobox gene 2(CDX2)signaling pathway in IM regulation.AIM To verify the regulation of the OLFM4/NOD1/CDX2 pathway in IM,specifically investigating WWD’s effectiveness on IM through this pathway.METHODS Immunohistochemistry for OLFM4,NOD1,and CDX2 was conducted on tissue microarray.GES-1 cells treated with chenodeoxycholic acid were utilized as IM cell models.OLFM4 short hairpin RNA(shRNA),NOD1 shRNA,and OLFM4 pcDNA were transfected to clarify the pathway regulatory relationships.Protein interactions were validated by coimmunoprecipitation.To explore WWD’s pharmacological actions,IM rat models were induced using N-methyl-N’-nitro-N-nitrosoguanidine followed by WWD gavage.Gastric cells were treated with WWD-medicated serum.Cytokines and chemokines content were assessed by enzyme-linked immunosorbent assay and quantitative reverse transcription polymerase chain reaction.RESULTS The OLFM4/NOD1/CDX2 axis was a characteristic of IM.OLFM4 exhibited direct binding and subsequent downregulation of NOD1,thereby sustaining the activation of CDX2 and promoting the progression of IM.WWD improved gastric mucosal histological lesions while suppressing intestinal markers KLF transcription factor 4,villin 1,and MUCIN 2 expression in IM rats.Regarding pharmacological actions,WWD suppressed OLFM4 and restored NOD1 expression,consequently reducing CDX2 at the mRNA and protein levels in IM rats.Parallel regulatory mechanisms were observed at the protein level in IM cells treated with WWD-medicated serum.Furthermore,WWD-medicated serum treatment strengthened OLFM4 and NOD1 interaction.In case of antiinflammatory,WWD restrained interleukin(IL)-6,interferon-gamma,IL-17,macrophage chemoattractant protein-1,macrophage inflammatory protein 1 alpha content in IM rat serum.WWD-medicated serum inhibited tumor necrosis factor alpha,IL-6,IL-8 transcriptions in IM cells.CONCLUSION The OLFM4/NOD1/CDX2 pathway is involved in the regulation of IM.WWD exerts its therapeutic efficacy on IM through the pathway,additionally attenuating the inflammatory response.展开更多
目的:研究胃萎方对信号转导及转录激活蛋白3(Signal Transducer and Activator of Transcription 3,STAT3)/核因子-κB(Nuclear Factor-κB,NF-κB)信号通路的调控作用,探讨胃萎方对胃癌前病变(Precancerous Lesion of Gastric Cancer,P...目的:研究胃萎方对信号转导及转录激活蛋白3(Signal Transducer and Activator of Transcription 3,STAT3)/核因子-κB(Nuclear Factor-κB,NF-κB)信号通路的调控作用,探讨胃萎方对胃癌前病变(Precancerous Lesion of Gastric Cancer,PLGC)的疗效及作用机制。方法:从70只雄性Wistar大鼠中随机选择20只作为空白组,其余50只为造模组。采用N-甲基-N’-硝基-N-亚硝基胍(MNNG)诱导GPL大鼠实验模型。14周后,对随机选择的造模组大鼠10只进行胃组织学分析,确认造模成功,造模组剩余40只大鼠随机分为模型组、胃萎方治疗组各20只。使用胃萎方干预后,观察大鼠体质量增长、胃黏膜组织结构变化,以及血清胃蛋白酶原(Pepsinogen,PG)Ⅰ,PGⅡ和胃泌素的浓度,并检测NF-κB p65和STAT3的基因及蛋白表达。结果:与空白组比较,模型组大鼠体质量增长缓慢,胃黏膜组织细胞异型增生明显,血清PGⅠ、胃泌素水平和PGⅠ/PGⅡ比值显著降低,胃黏膜NF-κB/STAT3信号通路被激活;与模型组比较,胃萎方治疗组大鼠整体情况改善,胃黏膜组织结构重塑、萎缩性胃黏膜和非典型腺上皮减少,PGⅠ/PGⅡ比值和胃泌素水平增加(P<0.01),NF-κB p65和STAT3的mRNA及蛋白的表达受到一定程度的抑制(P<0.05)。结论:胃萎方能够通过抑制NF-κB/STAT3信号通路逆转MNNG诱导的大鼠GPL,为胃萎方的临床应用提供了基础证据。展开更多
基金Supported by National Natural Science Foundation of China,No.82104747 and No.82305133Basic and Applied Basic Research Foundation of Guangdong Province,China,No.2020A1515110947+1 种基金General Project of Shenzhen Science and Technology Innovation Commission,No.JCYJ20230807094659030,No.JCYJ20230807094805012,No.JCYJ20210324111602007,and No.JCYJ20220531091815034the Scientific Research Project of Guangdong Provincial Bureau of Traditional Chinese Medicine,No.20221349.
文摘BACKGROUND Gastric intestinal metaplasia(IM)is a precancerous lesion that is associated with an elevated risk of gastric carcinogenesis.Weiwei Decoction(WWD)is a promising traditional Chinese herbal formula widely employed in clinical for treating IM.Previous studies suggested the potential involvement of the olfactomedin 4(OLFM4)/nucleotide-binding oligomerization domain 1(NOD1)/caudal-type homeobox gene 2(CDX2)signaling pathway in IM regulation.AIM To verify the regulation of the OLFM4/NOD1/CDX2 pathway in IM,specifically investigating WWD’s effectiveness on IM through this pathway.METHODS Immunohistochemistry for OLFM4,NOD1,and CDX2 was conducted on tissue microarray.GES-1 cells treated with chenodeoxycholic acid were utilized as IM cell models.OLFM4 short hairpin RNA(shRNA),NOD1 shRNA,and OLFM4 pcDNA were transfected to clarify the pathway regulatory relationships.Protein interactions were validated by coimmunoprecipitation.To explore WWD’s pharmacological actions,IM rat models were induced using N-methyl-N’-nitro-N-nitrosoguanidine followed by WWD gavage.Gastric cells were treated with WWD-medicated serum.Cytokines and chemokines content were assessed by enzyme-linked immunosorbent assay and quantitative reverse transcription polymerase chain reaction.RESULTS The OLFM4/NOD1/CDX2 axis was a characteristic of IM.OLFM4 exhibited direct binding and subsequent downregulation of NOD1,thereby sustaining the activation of CDX2 and promoting the progression of IM.WWD improved gastric mucosal histological lesions while suppressing intestinal markers KLF transcription factor 4,villin 1,and MUCIN 2 expression in IM rats.Regarding pharmacological actions,WWD suppressed OLFM4 and restored NOD1 expression,consequently reducing CDX2 at the mRNA and protein levels in IM rats.Parallel regulatory mechanisms were observed at the protein level in IM cells treated with WWD-medicated serum.Furthermore,WWD-medicated serum treatment strengthened OLFM4 and NOD1 interaction.In case of antiinflammatory,WWD restrained interleukin(IL)-6,interferon-gamma,IL-17,macrophage chemoattractant protein-1,macrophage inflammatory protein 1 alpha content in IM rat serum.WWD-medicated serum inhibited tumor necrosis factor alpha,IL-6,IL-8 transcriptions in IM cells.CONCLUSION The OLFM4/NOD1/CDX2 pathway is involved in the regulation of IM.WWD exerts its therapeutic efficacy on IM through the pathway,additionally attenuating the inflammatory response.
文摘目的:研究胃萎方对信号转导及转录激活蛋白3(Signal Transducer and Activator of Transcription 3,STAT3)/核因子-κB(Nuclear Factor-κB,NF-κB)信号通路的调控作用,探讨胃萎方对胃癌前病变(Precancerous Lesion of Gastric Cancer,PLGC)的疗效及作用机制。方法:从70只雄性Wistar大鼠中随机选择20只作为空白组,其余50只为造模组。采用N-甲基-N’-硝基-N-亚硝基胍(MNNG)诱导GPL大鼠实验模型。14周后,对随机选择的造模组大鼠10只进行胃组织学分析,确认造模成功,造模组剩余40只大鼠随机分为模型组、胃萎方治疗组各20只。使用胃萎方干预后,观察大鼠体质量增长、胃黏膜组织结构变化,以及血清胃蛋白酶原(Pepsinogen,PG)Ⅰ,PGⅡ和胃泌素的浓度,并检测NF-κB p65和STAT3的基因及蛋白表达。结果:与空白组比较,模型组大鼠体质量增长缓慢,胃黏膜组织细胞异型增生明显,血清PGⅠ、胃泌素水平和PGⅠ/PGⅡ比值显著降低,胃黏膜NF-κB/STAT3信号通路被激活;与模型组比较,胃萎方治疗组大鼠整体情况改善,胃黏膜组织结构重塑、萎缩性胃黏膜和非典型腺上皮减少,PGⅠ/PGⅡ比值和胃泌素水平增加(P<0.01),NF-κB p65和STAT3的mRNA及蛋白的表达受到一定程度的抑制(P<0.05)。结论:胃萎方能够通过抑制NF-κB/STAT3信号通路逆转MNNG诱导的大鼠GPL,为胃萎方的临床应用提供了基础证据。
