Objective To explore the pharmacodynamic mechanism of Fu Zi Li Zhong Decoction(FZLZD)in treating gastric ulcer and its effect on gastrointestinal microecology.Methods Rat gastric ulcer model was established by continu...Objective To explore the pharmacodynamic mechanism of Fu Zi Li Zhong Decoction(FZLZD)in treating gastric ulcer and its effect on gastrointestinal microecology.Methods Rat gastric ulcer model was established by continuous vinegar gavage.The experiment included normal group,model group,ranitidine group and FZLZD group.The animals were handled after intragastricing administration of corresponding clinical isodose drugs for 7 d.The rats’gastrointestinal histopathology was observed by HE staining.The expression of ZO-1 and Occludin mRNA in rat gastric tissue was detected by RT-PCR.The diversity of intestinal flora was analyzed by 16SrDNA sequencing.Results FZLZD can effectively improve gastric mucosal destruction,epithelial exfoliation,inflammatory cell infiltration,inflammatory substance exudation and edema in rats with gastric ulcer.FZLZD can effectively up-regulate the expression levels of ZO-1 and Occludin mRNA in gastric tissue of rats with gastric ulcer.The results of intestinal flora diversity showed that the diversity of intestinal flora in FZLZD group was significantly higher than that of model group.Conclusions FZLZD can regulate gastric ulcer by promoting the expression of ZO-1 and Occludin and improving the diversity of intestinal flora.展开更多
目的探究麻杏石甘汤通过外泌体miR-1249-5p靶向溶质载体家族4成员A1(solute carrier family 4 member 1,SLC4A1)抑制流感病毒诱导的肺上皮细胞损伤-巨噬细胞极化的级联损伤效应的作用机制。方法以肺上皮细胞为研究对象,采用CCK-8检测麻...目的探究麻杏石甘汤通过外泌体miR-1249-5p靶向溶质载体家族4成员A1(solute carrier family 4 member 1,SLC4A1)抑制流感病毒诱导的肺上皮细胞损伤-巨噬细胞极化的级联损伤效应的作用机制。方法以肺上皮细胞为研究对象,采用CCK-8检测麻杏石甘汤含药血清的安全剂量和对流感病毒刺激肺上皮细胞治疗剂量,NanoSight鉴定肺上皮细胞源外泌体的粒径和浓度,qRT-PCR检测外泌体中miR-1249-5p基因表达。以与肺上皮细胞源外泌体共培养的巨噬细胞为研究对象,采用qRT-PCR检测巨噬细胞中miR-1249-5p、SLC4A1和核因子-κB(nuclear factor-κB,NF-κB)通路相关基因表达;Western blotting检测巨噬细胞中SLC4A1和NF-κB通路相关蛋白表达;流式细胞术和qRT-PCR检测巨噬细胞M1/M2极化表型。结果CCK-8结果显示,10%和20%的麻杏石甘汤含药血清对肺上皮细胞无细胞毒性,显著抑制流感病毒诱导的肺上皮细胞损伤(P<0.05、0.01);NanoSight检测各组肺上皮细胞源外泌体大小无明显变化,外泌体浓度有所改变;qRT-PCR结果显示流感病毒刺激的肺上皮源外泌体miR-1249-5p基因表达明显降低(P<0.01),各给药组miR-1249-5p基因表达显著升高(P<0.05、0.001)。肺上皮源外泌体共培养后的巨噬细胞中miR-1249-5p基因表达与外泌体相一致(P<0.05、0.001);qRTPCR和Western blotting结果显示,模型组巨噬细胞SLC4A1和NF-κB通路基因和蛋白表达显著升高(P<0.01、0.001),各给药组SLC4A1和NF-κB通路基因和蛋白表达显著下降(P<0.05、0.01);流式细胞术结果显示模型组巨噬细胞CD86+比例明显增高(P<0.01),各给药组巨噬细胞CD86+比例显著降低(P<0.05);qRT-PCR结果显示共培养后模型组巨噬细胞肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)和IL-1β的表达显著升高(P<0.01、0.001),IL-10和转化生长因子-β1(transforming growth factor-β1,TGF-β1)的表达显著下降(P<0.05、0.01),各给药组以上因子表达趋势与之相反(P<0.05、0.01、0.001)。结论麻杏石甘汤可能通过肺上皮细胞源外泌体递送miR-1249-5p到巨噬细胞中,并靶向巨噬细胞中SLC4A1调控NF-κB信号通路,抑制流感病毒的细胞级联损伤效应的作用机制。展开更多
基金funding support from the National Natural Science Foundation of China (No. 81973670 and No. 81503445)Hunan Provincial Natural Science Foundation of China (No. 2016JJ2095)Hunan Province College Students Research Learning and Innovative Experiment Project (No. 2018-412)
文摘Objective To explore the pharmacodynamic mechanism of Fu Zi Li Zhong Decoction(FZLZD)in treating gastric ulcer and its effect on gastrointestinal microecology.Methods Rat gastric ulcer model was established by continuous vinegar gavage.The experiment included normal group,model group,ranitidine group and FZLZD group.The animals were handled after intragastricing administration of corresponding clinical isodose drugs for 7 d.The rats’gastrointestinal histopathology was observed by HE staining.The expression of ZO-1 and Occludin mRNA in rat gastric tissue was detected by RT-PCR.The diversity of intestinal flora was analyzed by 16SrDNA sequencing.Results FZLZD can effectively improve gastric mucosal destruction,epithelial exfoliation,inflammatory cell infiltration,inflammatory substance exudation and edema in rats with gastric ulcer.FZLZD can effectively up-regulate the expression levels of ZO-1 and Occludin mRNA in gastric tissue of rats with gastric ulcer.The results of intestinal flora diversity showed that the diversity of intestinal flora in FZLZD group was significantly higher than that of model group.Conclusions FZLZD can regulate gastric ulcer by promoting the expression of ZO-1 and Occludin and improving the diversity of intestinal flora.
