Intracerebral hemorrhage is the most dangerous subtype of stroke,characterized by high mortality and morbidity rates,and frequently leads to significant secondary white matter injury.In recent decades,studies have rev...Intracerebral hemorrhage is the most dangerous subtype of stroke,characterized by high mortality and morbidity rates,and frequently leads to significant secondary white matter injury.In recent decades,studies have revealed that gut microbiota can communicate bidirectionally with the brain through the gut microbiota–brain axis.This axis indicates that gut microbiota is closely related to the development and prognosis of intracerebral hemorrhage and its associated secondary white matter injury.The NACHT,LRR,and pyrin domain-containing protein 3(NLRP3)inflammasome plays a crucial role in this context.This review summarizes the dysbiosis of gut microbiota following intracerebral hemorrhage and explores the mechanisms by which this imbalance may promote the activation of the NLRP3 inflammasome.These mechanisms include metabolic pathways(involving short-chain fatty acids,lipopolysaccharides,lactic acid,bile acids,trimethylamine-N-oxide,and tryptophan),neural pathways(such as the vagus nerve and sympathetic nerve),and immune pathways(involving microglia and T cells).We then discuss the relationship between the activated NLRP3 inflammasome and secondary white matter injury after intracerebral hemorrhage.The activation of the NLRP3 inflammasome can exacerbate secondary white matter injury by disrupting the blood–brain barrier,inducing neuroinflammation,and interfering with nerve regeneration.Finally,we outline potential treatment strategies for intracerebral hemorrhage and its secondary white matter injury.Our review highlights the critical role of the gut microbiota–brain axis and the NLRP3 inflammasome in white matter injury following intracerebral hemorrhage,paving the way for exploring potential therapeutic approaches.展开更多
BACKGROUND Chronic atrophic gastritis(CAG)is a clinically refractory gastric disease often characterized by high recurrence rates and adverse drug reactions.Anwei decoction(AWD),a traditional Chinese medicine formula,...BACKGROUND Chronic atrophic gastritis(CAG)is a clinically refractory gastric disease often characterized by high recurrence rates and adverse drug reactions.Anwei decoction(AWD),a traditional Chinese medicine formula,has been shown to significantly improve clinical symptoms in patients with CAG,as demonstrated by a multicenter cohort study(overall effective rate:82.5%,P<0.01).However,the unclear molecular mechanisms and therapeutic targets of AWD limit its international acceptance.AIM To investigate the therapeutic mechanisms of AWD against CAG from an integrated perspective.METHODS In this study,N-methyl-N’-nitro-N-nitrosoguanidine was used to establish a CAG rat model.Serum-derived constituents transferred from AWD were first identified using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry.The concentrations of inflammatory cytokines in serum samples were determined by enzyme-linked immunosorbent assay.Moreover,gastric mucosal tissues were analyzed by quantitative realtime polymerase chain reaction to measure messenger RNA(mRNA)levels of the NLRP3 inflammasome.Western blotting was used to detect the protein expression of NLRP3,caspase-1,and interleukin(IL)-1β.To elucidate the regulatory mechanisms underlying AWD treatment,structural alterations of the gut microbiota(GM)and associated metabolites were analyzed using integrated high-throughput sequencing(16S rRNA)and liquid chromatography-mass spectrometry based untargeted metabolomics.This comprehensive approach systematically clarified AWD’s multi-target therapeutic mechanisms against CAG.RESULTS AWD notably reduced serum levels of pro-inflammatory cytokines,such as IL-1β,IL-18,tumor necrosis factor-α,and lipopolysaccharide,demonstrating significant statistical differences(all P<0.01).Additionally,AWD substantially inhibited NLRP3 mRNA expression in gastric mucosal tissue(P<0.01)and concurrently decreased the protein abundance of NLRP3,IL-1β,and caspase-1(all P<0.01),thereby suppressing inflammasome signaling activation.GM analysis indicated that AWD intervention significantly increased the relative abundance of beneficial bacteria.Associated microbial metabolites likely inhibited the NLRP3 inflammasome pathway by modulating immune cell function.Non-targeted metabolomics further indicated that AWD exerted anti-inflammatory effects by regulating critical metabolic pathways,including the Kaposi’s sarcoma-associated herpesvirus infection pathway,autophagy processes,and glycosylphosphatidylinositol-anchor biosynthesis.CONCLUSION AWD alleviates the pathological progression of CAG through multi-target synergistic mechanisms.On one hand,AWD directly suppresses gastric mucosal inflammation by inhibiting NLRP3 inflammasome activation.On the other hand,AWD remodels intestinal microbiota-metabolite homeostasis,enhances intestinal barrier function,and regulates mucosal immune responses.展开更多
目的探讨血管软化丸能否通过调控腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)/NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)通路抑制细胞焦亡改善载脂蛋白E基因...目的探讨血管软化丸能否通过调控腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)/NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)通路抑制细胞焦亡改善载脂蛋白E基因敲除(apolipoprotein E knockout,APOE^(-/-))小鼠动脉粥样硬化(atherosclerosis,AS)。方法10只C57BL/6J小鼠作为空白对照组,60只APOE^(-/-)小鼠随机分为模型组、血管软化丸低剂量组、血管软化丸高剂量组、阿托伐他汀组、血管软化丸高剂量和AMPK抑制剂联合组、AMPK抑制剂组,给予高脂饮食饲养18周建造动脉粥样硬化小鼠模型,并于第13周给予不同方法干预,干预6周后取材,分别检测各组小鼠甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)、低密度脂蛋白-胆固醇(low density lipoprotein-eholesterol,LDL-C)、高密度脂蛋白-胆固醇(high density lipoprotein-eholesterol,HDL-C)水平,HE和油红O染色观察小鼠主动脉组织病理形态学变化,Elisa测定主动脉组织白细胞介素(interleukin,IL)-1β和IL-18水平,Western Blot检测AMPK、P-AMPK、NLRP3、裂解的半胱氨酸天冬酶1(Cleaved-Caspase-1)、Gasdermin D(GSDMD)-N蛋白表达,免疫荧光检测P-AMPK、NLRP3蛋白在主动脉根部的分布情况,透射电子显微镜观察主动脉内皮超微结构改变情况。结果与对照组相比,模型组大鼠血清TG、TC、LDL-C水平显著升高,HDL-C水平显著降低(P<0.01),主动脉根部细胞排列紊乱,且脂肪空泡明显,主动脉有明显粥样斑块沉积,P-AMPK蛋白表达水平显著降低,NLRP3、Cleaved-Caspase 1、GSDMD-N蛋白表达水平及IL-1β和IL-18水平显著升高(P<0.05或P<0.01);与模型组比较,血管软化丸高低剂量组及阿托伐他汀组血脂紊乱得到改善,主动脉组织及细胞病变程度减轻,P-AMPK蛋白表达水平显著升高,NLRP3、Cleaved-Caspase 1、GSDMD-N蛋白表达水平显著降低(P<0.05或P<0.01),IL-1β和IL-18水平显著降低(P<0.05或P<0.01);与血管软化丸高剂量+AMPK抑制剂组比较,血管软化丸高剂量组焦亡相关蛋白水平显著降低(P<0.05或P<0.01),主动脉内皮细胞损伤程度减轻,而AMPK抑制剂组与之相反。结论血管软化丸可能通过调控AMPK/NLRP3通路减轻细胞焦亡发挥对动脉粥样硬化小鼠的保护作用。展开更多
基金supported by the Guangdong Basic and Applied Basic Research Foundation,No.2023A1515030045(to HS)Presidential Foundation of Zhujiang Hospital of Southern Medical University,No.yzjj2022ms4(to HS)。
文摘Intracerebral hemorrhage is the most dangerous subtype of stroke,characterized by high mortality and morbidity rates,and frequently leads to significant secondary white matter injury.In recent decades,studies have revealed that gut microbiota can communicate bidirectionally with the brain through the gut microbiota–brain axis.This axis indicates that gut microbiota is closely related to the development and prognosis of intracerebral hemorrhage and its associated secondary white matter injury.The NACHT,LRR,and pyrin domain-containing protein 3(NLRP3)inflammasome plays a crucial role in this context.This review summarizes the dysbiosis of gut microbiota following intracerebral hemorrhage and explores the mechanisms by which this imbalance may promote the activation of the NLRP3 inflammasome.These mechanisms include metabolic pathways(involving short-chain fatty acids,lipopolysaccharides,lactic acid,bile acids,trimethylamine-N-oxide,and tryptophan),neural pathways(such as the vagus nerve and sympathetic nerve),and immune pathways(involving microglia and T cells).We then discuss the relationship between the activated NLRP3 inflammasome and secondary white matter injury after intracerebral hemorrhage.The activation of the NLRP3 inflammasome can exacerbate secondary white matter injury by disrupting the blood–brain barrier,inducing neuroinflammation,and interfering with nerve regeneration.Finally,we outline potential treatment strategies for intracerebral hemorrhage and its secondary white matter injury.Our review highlights the critical role of the gut microbiota–brain axis and the NLRP3 inflammasome in white matter injury following intracerebral hemorrhage,paving the way for exploring potential therapeutic approaches.
基金Supported by the National Natural Science Foundation of China,No.81860843Guangxi Administration of Traditional Chinese Medicine Project,No.GZSY23-36 and No.GXZYA20240150。
文摘BACKGROUND Chronic atrophic gastritis(CAG)is a clinically refractory gastric disease often characterized by high recurrence rates and adverse drug reactions.Anwei decoction(AWD),a traditional Chinese medicine formula,has been shown to significantly improve clinical symptoms in patients with CAG,as demonstrated by a multicenter cohort study(overall effective rate:82.5%,P<0.01).However,the unclear molecular mechanisms and therapeutic targets of AWD limit its international acceptance.AIM To investigate the therapeutic mechanisms of AWD against CAG from an integrated perspective.METHODS In this study,N-methyl-N’-nitro-N-nitrosoguanidine was used to establish a CAG rat model.Serum-derived constituents transferred from AWD were first identified using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry.The concentrations of inflammatory cytokines in serum samples were determined by enzyme-linked immunosorbent assay.Moreover,gastric mucosal tissues were analyzed by quantitative realtime polymerase chain reaction to measure messenger RNA(mRNA)levels of the NLRP3 inflammasome.Western blotting was used to detect the protein expression of NLRP3,caspase-1,and interleukin(IL)-1β.To elucidate the regulatory mechanisms underlying AWD treatment,structural alterations of the gut microbiota(GM)and associated metabolites were analyzed using integrated high-throughput sequencing(16S rRNA)and liquid chromatography-mass spectrometry based untargeted metabolomics.This comprehensive approach systematically clarified AWD’s multi-target therapeutic mechanisms against CAG.RESULTS AWD notably reduced serum levels of pro-inflammatory cytokines,such as IL-1β,IL-18,tumor necrosis factor-α,and lipopolysaccharide,demonstrating significant statistical differences(all P<0.01).Additionally,AWD substantially inhibited NLRP3 mRNA expression in gastric mucosal tissue(P<0.01)and concurrently decreased the protein abundance of NLRP3,IL-1β,and caspase-1(all P<0.01),thereby suppressing inflammasome signaling activation.GM analysis indicated that AWD intervention significantly increased the relative abundance of beneficial bacteria.Associated microbial metabolites likely inhibited the NLRP3 inflammasome pathway by modulating immune cell function.Non-targeted metabolomics further indicated that AWD exerted anti-inflammatory effects by regulating critical metabolic pathways,including the Kaposi’s sarcoma-associated herpesvirus infection pathway,autophagy processes,and glycosylphosphatidylinositol-anchor biosynthesis.CONCLUSION AWD alleviates the pathological progression of CAG through multi-target synergistic mechanisms.On one hand,AWD directly suppresses gastric mucosal inflammation by inhibiting NLRP3 inflammasome activation.On the other hand,AWD remodels intestinal microbiota-metabolite homeostasis,enhances intestinal barrier function,and regulates mucosal immune responses.
文摘目的探讨血管软化丸能否通过调控腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)/NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)通路抑制细胞焦亡改善载脂蛋白E基因敲除(apolipoprotein E knockout,APOE^(-/-))小鼠动脉粥样硬化(atherosclerosis,AS)。方法10只C57BL/6J小鼠作为空白对照组,60只APOE^(-/-)小鼠随机分为模型组、血管软化丸低剂量组、血管软化丸高剂量组、阿托伐他汀组、血管软化丸高剂量和AMPK抑制剂联合组、AMPK抑制剂组,给予高脂饮食饲养18周建造动脉粥样硬化小鼠模型,并于第13周给予不同方法干预,干预6周后取材,分别检测各组小鼠甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)、低密度脂蛋白-胆固醇(low density lipoprotein-eholesterol,LDL-C)、高密度脂蛋白-胆固醇(high density lipoprotein-eholesterol,HDL-C)水平,HE和油红O染色观察小鼠主动脉组织病理形态学变化,Elisa测定主动脉组织白细胞介素(interleukin,IL)-1β和IL-18水平,Western Blot检测AMPK、P-AMPK、NLRP3、裂解的半胱氨酸天冬酶1(Cleaved-Caspase-1)、Gasdermin D(GSDMD)-N蛋白表达,免疫荧光检测P-AMPK、NLRP3蛋白在主动脉根部的分布情况,透射电子显微镜观察主动脉内皮超微结构改变情况。结果与对照组相比,模型组大鼠血清TG、TC、LDL-C水平显著升高,HDL-C水平显著降低(P<0.01),主动脉根部细胞排列紊乱,且脂肪空泡明显,主动脉有明显粥样斑块沉积,P-AMPK蛋白表达水平显著降低,NLRP3、Cleaved-Caspase 1、GSDMD-N蛋白表达水平及IL-1β和IL-18水平显著升高(P<0.05或P<0.01);与模型组比较,血管软化丸高低剂量组及阿托伐他汀组血脂紊乱得到改善,主动脉组织及细胞病变程度减轻,P-AMPK蛋白表达水平显著升高,NLRP3、Cleaved-Caspase 1、GSDMD-N蛋白表达水平显著降低(P<0.05或P<0.01),IL-1β和IL-18水平显著降低(P<0.05或P<0.01);与血管软化丸高剂量+AMPK抑制剂组比较,血管软化丸高剂量组焦亡相关蛋白水平显著降低(P<0.05或P<0.01),主动脉内皮细胞损伤程度减轻,而AMPK抑制剂组与之相反。结论血管软化丸可能通过调控AMPK/NLRP3通路减轻细胞焦亡发挥对动脉粥样硬化小鼠的保护作用。
