目的探讨神经生长因子(NGF)及其高亲和力受体酪氨酸激酶A(TrkA)在支气管哮喘(BA)发病中的作用,评估其与哮喘严重程度及2型辅助T(Th2)炎症因子的相关性,并验证基于聚乳酸-羟基乙酸共聚物(PLGA)微球负载抗NGF抗体的雾化靶向治疗在重症哮...目的探讨神经生长因子(NGF)及其高亲和力受体酪氨酸激酶A(TrkA)在支气管哮喘(BA)发病中的作用,评估其与哮喘严重程度及2型辅助T(Th2)炎症因子的相关性,并验证基于聚乳酸-羟基乙酸共聚物(PLGA)微球负载抗NGF抗体的雾化靶向治疗在重症哮喘中的干预效果及机制。方法收集不同严重程度哮喘患者与非哮喘人群的血清,检测NGF、TrkA及白细胞介素4(IL-4)、IL-5、IL-13水平。构建卵清蛋白(OVA)诱导的重症哮喘小鼠模型,随机分为空白对照组、重症哮喘组、常规抗体治疗组和雾化抗NGF微球组,通过ELISA、肺功能测定、HE/Masson染色和Western blot法比较各组间支气管肺泡灌洗液(BALF)中炎症因子水平、肺组织病理变化及NGF/TrkA/核因子κB(NF-κB)蛋白表达水平差异。结果临床检测中,重度哮喘组患者较非哮喘人群对照组血清中NGF(82.52±4.01 vs 22.92±5.25)pg/mL和TrkA(53.93±8.97 vs 15.76±3.60)pg/mL水平升高;IL-4(44.23±9.88 vs 20.88±3.90)pg/mL、IL-5(147.1±14.92 vs 47.13±5.63)pg/mL、IL-13(106.9±11.64 vs 51.93±8.55)pg/mL水平显著升高。动物实验中,雾化抗NGF微球组较重症哮喘组小鼠BALF中IL-4(36.08±6.08 vs 87.35±9.13)pg/mL、IL-5(35.41±4.52 vs 81.53±4.94)pg/mL、IL-13(36.37±4.57 vs 92.37±6.53)pg/mL水平降低,第一秒用力呼气容积(FEV1)(84.68±5.33 vs 60.87±3.93)%升高、气道高反应性(AHR)(152.8±7.36 vs 233.8±8.77)%下降,肺组织病理损伤与胶原沉积减轻。Western blot法显示,雾化抗NGF微球组小鼠肺组织中NGF、TrkA和NF-κB相对蛋白水平较重症哮喘组小鼠低。结论NGF/TrkA信号通路与哮喘严重程度密切相关,雾化抗NGF微球治疗可有效靶向肺部,抑制NF-κB通路,缓解气道炎症,显著改善肺功能。展开更多
目的 :寻找新的 Trk A可能底物或调控蛋白 ,以深入认识 Trk A下游信号转导机制。方法 :将 Trk A胞内域与 L ex A蛋白融合作为诱饵蛋白 ,应用酵母双杂交方法筛选人脑 L ex A c DNA文库 ,经β-半乳糖苷酶活性鉴定和测序分析进一步鉴别 ,...目的 :寻找新的 Trk A可能底物或调控蛋白 ,以深入认识 Trk A下游信号转导机制。方法 :将 Trk A胞内域与 L ex A蛋白融合作为诱饵蛋白 ,应用酵母双杂交方法筛选人脑 L ex A c DNA文库 ,经β-半乳糖苷酶活性鉴定和测序分析进一步鉴别 ,获得阳性克隆。 结果 :明确阳性克隆中的 Trk AIC- BP1为一种新的 Trk A结合蛋白。免疫共沉淀实验证实了 Trk AIC- BP1和Trk A在酵母中的相互作用。结论 :首次筛选到 Trk A结合蛋白—— Trk AIC- BP1,它可能在 Trk展开更多
Irritable bowel syndrome-diarrhea(IBS-D)is a prevalent functional bowel disease severely impairs the quality of life of patients.Visceral hypersensitivity(VH)is the main symptom of IBS-D.Probiotics have been found to ...Irritable bowel syndrome-diarrhea(IBS-D)is a prevalent functional bowel disease severely impairs the quality of life of patients.Visceral hypersensitivity(VH)is the main symptom of IBS-D.Probiotics have been found to effectively alleviate VH.In this study,neonatal maternal separation(NMS)was used to induce VH in 72 rats and investigated the effects of daily gavage of 3×10^(9) CFU of Lactobacillus and Bifidobacterium on alleviating the symptoms of IBS-D and behavioral interventions,as well as exploring potential mechanisms.The results demonstrated that L.acidophilus and B.breve could alleviate the symptoms of IBS-D such as VH and increased frequency of bowel movements,as well as improving non-spatial memory in NMS rats.These effects were achieved by modulating the composition of gut microbiota and metabolites(including short-chain fatty acids,deoxycholic acid and adenosine),and by regulating the expression of genes and proteins associated with the NGF/TrkA signaling pathway,tryptophan metabolism,and inflammatory factors in the hippocampus.This finding suggests that L.acidophilus and B.breve have promising potential for the alleviation of IBS.展开更多
目的:研究子宫内膜异位症(内异症)患者在位内膜神经生长因子(NGF)及其受体trkA、p75NTR的表达,探讨上述因子与内异症疼痛的关系。方法:选择就诊于北京协和医院并进行了腹腔镜手术的28例子宫内膜异位症患者作为研究组,非子宫内膜异位症患...目的:研究子宫内膜异位症(内异症)患者在位内膜神经生长因子(NGF)及其受体trkA、p75NTR的表达,探讨上述因子与内异症疼痛的关系。方法:选择就诊于北京协和医院并进行了腹腔镜手术的28例子宫内膜异位症患者作为研究组,非子宫内膜异位症患者10例作为对照组(B组),收集上述患者分泌期在位子宫内膜。根据患者有无痛经,分为内异症疼痛组(A1组,18例)及内异症非疼痛组(A2组,10例)。采用免疫组化比较各组病灶中NGF及其受体trkA、p75NTR的表达,并分析其与痛经的关系。结果:各组在位内膜腺上皮中NGF的表达显著高于间质(P<0.05);内异症疼痛组腺上皮NGF表达较非疼痛组明显升高(359.9±18.7 vs 201.3±34.3,P<0.05)。p75NTR主要在子宫内膜间质细胞中表达;内异症组明显高于非内异症组(58.8±21.1、22.5±16.1 vs 0,P<0.05);内异症疼痛组较非疼痛组p75NTR表达明显升高(58.8±21.1 vs 22.5±16.1,P<0.05)。trkA在非内异症组子宫内膜间质中表达明显高于腺上皮(P<0.05);在内异症组腺上皮中,trkA的表达较非内异症组明显增高(P<0.05);trkA的表达量与内异症患者疼痛无关。结论:p75NTR可能参与内异症的发病,NGF及其受体p75NTR在在位内膜中的表达与患者疼痛相关,表明其可能参与了内异症疼痛发病机制。展开更多
文摘目的探讨神经生长因子(NGF)及其高亲和力受体酪氨酸激酶A(TrkA)在支气管哮喘(BA)发病中的作用,评估其与哮喘严重程度及2型辅助T(Th2)炎症因子的相关性,并验证基于聚乳酸-羟基乙酸共聚物(PLGA)微球负载抗NGF抗体的雾化靶向治疗在重症哮喘中的干预效果及机制。方法收集不同严重程度哮喘患者与非哮喘人群的血清,检测NGF、TrkA及白细胞介素4(IL-4)、IL-5、IL-13水平。构建卵清蛋白(OVA)诱导的重症哮喘小鼠模型,随机分为空白对照组、重症哮喘组、常规抗体治疗组和雾化抗NGF微球组,通过ELISA、肺功能测定、HE/Masson染色和Western blot法比较各组间支气管肺泡灌洗液(BALF)中炎症因子水平、肺组织病理变化及NGF/TrkA/核因子κB(NF-κB)蛋白表达水平差异。结果临床检测中,重度哮喘组患者较非哮喘人群对照组血清中NGF(82.52±4.01 vs 22.92±5.25)pg/mL和TrkA(53.93±8.97 vs 15.76±3.60)pg/mL水平升高;IL-4(44.23±9.88 vs 20.88±3.90)pg/mL、IL-5(147.1±14.92 vs 47.13±5.63)pg/mL、IL-13(106.9±11.64 vs 51.93±8.55)pg/mL水平显著升高。动物实验中,雾化抗NGF微球组较重症哮喘组小鼠BALF中IL-4(36.08±6.08 vs 87.35±9.13)pg/mL、IL-5(35.41±4.52 vs 81.53±4.94)pg/mL、IL-13(36.37±4.57 vs 92.37±6.53)pg/mL水平降低,第一秒用力呼气容积(FEV1)(84.68±5.33 vs 60.87±3.93)%升高、气道高反应性(AHR)(152.8±7.36 vs 233.8±8.77)%下降,肺组织病理损伤与胶原沉积减轻。Western blot法显示,雾化抗NGF微球组小鼠肺组织中NGF、TrkA和NF-κB相对蛋白水平较重症哮喘组小鼠低。结论NGF/TrkA信号通路与哮喘严重程度密切相关,雾化抗NGF微球治疗可有效靶向肺部,抑制NF-κB通路,缓解气道炎症,显著改善肺功能。
基金supported by the National Key Research and Development Program of China(2021YFD2100700)the National Natural Science Foundation of China(32021005)+1 种基金the 111 project(BP0719028)the Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province.
文摘Irritable bowel syndrome-diarrhea(IBS-D)is a prevalent functional bowel disease severely impairs the quality of life of patients.Visceral hypersensitivity(VH)is the main symptom of IBS-D.Probiotics have been found to effectively alleviate VH.In this study,neonatal maternal separation(NMS)was used to induce VH in 72 rats and investigated the effects of daily gavage of 3×10^(9) CFU of Lactobacillus and Bifidobacterium on alleviating the symptoms of IBS-D and behavioral interventions,as well as exploring potential mechanisms.The results demonstrated that L.acidophilus and B.breve could alleviate the symptoms of IBS-D such as VH and increased frequency of bowel movements,as well as improving non-spatial memory in NMS rats.These effects were achieved by modulating the composition of gut microbiota and metabolites(including short-chain fatty acids,deoxycholic acid and adenosine),and by regulating the expression of genes and proteins associated with the NGF/TrkA signaling pathway,tryptophan metabolism,and inflammatory factors in the hippocampus.This finding suggests that L.acidophilus and B.breve have promising potential for the alleviation of IBS.
文摘目的:研究子宫内膜异位症(内异症)患者在位内膜神经生长因子(NGF)及其受体trkA、p75NTR的表达,探讨上述因子与内异症疼痛的关系。方法:选择就诊于北京协和医院并进行了腹腔镜手术的28例子宫内膜异位症患者作为研究组,非子宫内膜异位症患者10例作为对照组(B组),收集上述患者分泌期在位子宫内膜。根据患者有无痛经,分为内异症疼痛组(A1组,18例)及内异症非疼痛组(A2组,10例)。采用免疫组化比较各组病灶中NGF及其受体trkA、p75NTR的表达,并分析其与痛经的关系。结果:各组在位内膜腺上皮中NGF的表达显著高于间质(P<0.05);内异症疼痛组腺上皮NGF表达较非疼痛组明显升高(359.9±18.7 vs 201.3±34.3,P<0.05)。p75NTR主要在子宫内膜间质细胞中表达;内异症组明显高于非内异症组(58.8±21.1、22.5±16.1 vs 0,P<0.05);内异症疼痛组较非疼痛组p75NTR表达明显升高(58.8±21.1 vs 22.5±16.1,P<0.05)。trkA在非内异症组子宫内膜间质中表达明显高于腺上皮(P<0.05);在内异症组腺上皮中,trkA的表达较非内异症组明显增高(P<0.05);trkA的表达量与内异症患者疼痛无关。结论:p75NTR可能参与内异症的发病,NGF及其受体p75NTR在在位内膜中的表达与患者疼痛相关,表明其可能参与了内异症疼痛发病机制。
