增液通腑逐瘀方调控cGAS-STING信号通路改善脓毒症大鼠肠道通透性作用机制OA
Mechanism of Zengye Tongfu Zhuyu Prescription in Regulating cGAS-STING Signaling Pathway to Improve Intestinal Permeability in Septic Rats
目的 研究增液通腑逐瘀方对脓毒症大鼠肠道通透性的改善作用.方法 采用盲肠结扎穿孔术(Cecal ligation and puncture,CLP)建立脓毒症大鼠模型,将36只健康SPF级雄性SD大鼠,采用随机数字表法分为假手术组、模型组、增液通腑逐瘀方低剂量组、增液通腑逐瘀方高剂量组,每组9只.增液通腑逐瘀方低、高剂量组大鼠分别给予增液通腑逐瘀方0.5 g/ml、1 g/ml灌胃,假手术组及模型组给予同体积生理盐水灌胃,连续给药3 d后处死,留取回肠组织,制备病理学切片.采用Chiu's评分测定肠道损伤程度;采用蛋白质免疫印迹法(Western blot)检测肠道环鸟苷酸腺苷酸合酶(Cyclic GMP-AMP synthase,cGAS)、干扰素基因刺激因子(Stimulator of interferon genes,STING)、肌球轻链激酶(Myosin light chain kinase,MLCK)、闭合蛋白(Occludin)和闭锁小带蛋白-1(Zonula occludens-1,ZO-1)表达水平;采用实时定量反转录聚合酶连锁反应法(RT-qPCR)检测肠道肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)和干扰素-α(Interferon-α,IFN-α)的信使核糖核酸(Messenger ribonucleic acid,mRNA)表达水平;采用内毒素(Lipopolysaccharide,LPS)法检测肠道通透性.结果 与假手术组相比,模型组Chiu's评分,cGAS、STING、MLCK相对表达量,TNF-α和IFN-α的mRNA相对表达量及LPS含量均显著升高,差异有统计学意义(P<0.01),Occludin及ZO-1相对表达量均显著降低,差异有统计学意义(P<0.01).与模型组相比,增液通腑逐瘀方低剂量组Chiu's评分、MLCK相对表达量、IFN-α的mRNA相对表达量及LPS含量显著降低,差异有统计学意义(P<0.05,P<0.01),ZO-1相对表达量显著升高,差异有统计学意义(P<0.01);增液通腑逐瘀方高剂量组Chiu's评分,cGAS、STING、MLCK相对表达量,TNF-α和IFN-α的mRNA相对表达量及LPS含量均显著降低,差异有统计学意义(P<0.01),而Occludin及ZO-1相对表达量均显著升高,差异有统计学意义(P<0.01).结论 增液通腑逐瘀方可抑制cGAS-STING通路激活,抑制Ⅰ型干扰素(Interferon-Ⅰ,IFN-Ⅰ)反应及炎性介质释放,减少肠道紧密连接(Tight junctions,TJs)改变,改善肠道通透性,维持肠黏膜屏障完整性.
Objective To study the improvement of intestinal permeability in septic rats by Zengye Tongfu Zhuyu Prescription.Methods The model of septic rats was established by cecal ligation and puncture(CLP).A total of 36 healthy SPF grade male SD rats were randomly divided into a sham group,a model group,and low-and high-dose Zengye Tongfu Zhuyu Prescription groups using a random number table method,with 9 rats in each group.After 3 days of continuous administration,they were sacrificed,and ileal tissues were collected and prepared for pathological sections.The degree of intestinal injury was determined using the Chiu's score.Western blot was employed to detect the expression of intestinal cyclic GMP-AMP synthase(cGAS),stimulator of interferon genes(STING),myosin light chain kinase(MLCK),occludin,and zonula occludens-1(ZO-1).The messenger ribonucleic acid(mRNA)expression of intestinal tumor necrosis factor-α(TNF-α)and interferon-α(IFN-α)was measured by real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR).Intestinal permeability was measured by the lipopolysaccharide(LPS)method.Results Compared with the sham group,the model group showed significantly increased Chiu's score,relative expression of cGAS,STING,and MLCK,and mRNA relative expression of TNF-α and IFN-α(P<0.01)and significantly decreased relative expression of occludin and ZO-1(P<0.01).Compared to the model group,the low-dose Zengye Tongfu Zhuyu Prescription group exhibited significantly decreased Chiu's score,relative expression of MLCK,mRNA relative expression of IFN-α,and LPS content(P<0.05 or P<0.01)and significantly increased relative expression level of ZO-1(P<0.01).As for the high-dose Zengye Tongfu Zhuyu Prescription group,the Chiu's score,relative expression of cGAS,STING,and MLCK,mRNA relative expression of TNF-α and IFN-α,and LPS content were significantly decreased(P<0.01),while the relative expression of occludin and ZO-1 was significantly increased(P<0.01).Conclusion The Zengye Tongfu Zhuyu Prescription can inhibit the activation of the cGAS-STING pathway,suppress type I interferon(IFN-I)response and the release of inflammatory mediators,and reduce the change of intestinal tight junctions(TJs),thereby improving intestinal permeability and maintaining the integrity of intestinal mucosal barrier.
高薇薇;高万朋;关鹏
天津中医药大学第二附属医院急诊科,天津 300250天津中医药大学第二附属医院急诊科,天津 300250天津中医药大学第二附属医院急诊科,天津 300250
医药卫生
脓毒症增液通腑逐瘀方肠道通透性cGAS-STING信号通路肠上皮细胞
sepsisZengye Tongfu Zhuyu Prescriptionintestinal permeabilitycGAS-STING signaling pathwayintestinal epithelial cells
《世界中西医结合杂志》 2026 (2)
258-264,7
天津市教委科研计划项目(自然科学)(2022KJ185)天津市卫生健康科技项目(TJWJ2024MS014)
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