AIM:To investigate the effects of a Chinese medicine formula“Qingxuan Runmu Yin”(QRY)on ocular surface inflammation in a rat model of dry eye,and its mechanism via the toll-like receptor 4(TLR4)/transforming growth ...AIM:To investigate the effects of a Chinese medicine formula“Qingxuan Runmu Yin”(QRY)on ocular surface inflammation in a rat model of dry eye,and its mechanism via the toll-like receptor 4(TLR4)/transforming growth factor kinase 1(TAK1)/p38 mitogen-activated protein kinase(p38MAPK)signaling pathway.METHODS:Seventy-two Sprague-Dawley rats were randomly divided into six groups(n=12 each):the control group,model group,3 groups of QRY(with low-,medium-,and high-doses),and SB203580 group.Dry eye was induced using benzalkonium chloride.Schirmer’s test(SIT)and corneal fluorescein staining(CFS)were performed every 14d throughout the experiment.Histopathological changes in corneal and conjunctival tissues were observed using hematoxylin and eosin(HE)and periodic acid-Schiff(PAS)staining.Protein expression levels of key inflammatory markers and signaling pathway targets were assessed via immunohistochemistry,ELISA,and Western blotting.RESULTS:Compared to the control group,the model group showed significant reductions in SIT and increases in CFS scores,alongside structural disorganization of corneal/conjunctival tissues,decreased conjunctival goblet cell(CGC)numbers,and elevated expression of inflammatory markers[interleukin(IL)-1β,IL-6,tumor necrosis factoralpha(TNF-α),matrix metalloproteinase-9(MMP9)]and pathway proteins(TLR4,p-TAK1,p-p38MAPK;P<0.05).Treatment with QRY(low,medium,and high doses)and SB203580 significantly improved SIT scores,reduced CFS scores,restored corneoconjunctival structure,increased CGC numbers,and decreased expression levels of IL-1β,IL-6,TNF-α,MMP9,TLR4,p-TAK1,and p-p38MAPK proteins compared to the model group(P<0.05).CONCLUSION:QRY may alleviate ocular surface inflammation associated with dry eye by inhibiting the TLR4/TAK1/p38MAPK signaling pathway,highlighting its potential therapeutic efficacy for dry eye.展开更多
目的:探讨氧化苦参碱(OMT)联合复方茵陈颗粒(FYK)对脂多糖(LPS)诱导的大鼠肝BRL-3A细胞凋亡的抑制作用及其机制,为该联合用药抑制肝细胞凋亡提供更多的实验依据。方法:采用LPS诱导大鼠肝BRL-3A细胞建立细胞凋亡模型。应用CCK-8法测定BRL...目的:探讨氧化苦参碱(OMT)联合复方茵陈颗粒(FYK)对脂多糖(LPS)诱导的大鼠肝BRL-3A细胞凋亡的抑制作用及其机制,为该联合用药抑制肝细胞凋亡提供更多的实验依据。方法:采用LPS诱导大鼠肝BRL-3A细胞建立细胞凋亡模型。应用CCK-8法测定BRL-3A细胞活力;流式细胞术检测细胞凋亡率;Western blot法检测BRL-3A细胞中TLR4、P38、P-P38、JNK及P-JNK蛋白的表达;RT-q PCR法检测BRL-3A细胞中Bax、Bcl-2及Caspase-3 m RNA的表达。结果:CCK-8法结果显示,OMT可以减弱由LPS引起的细胞活力降低(P<0.05),OMT+FYK-M+LPS组及OMT+FYK-H+LPS组细胞活力高于OMT组(P<0.05);OMT预处理后细胞凋亡率低于LPS组(P<0.05),OMT+FYK-M+LPS组及OMT+FYK-H+LPS组细胞凋亡率低于OMT组(P<0.05)。Western blot结果显示,LPS组TLR4、P-P38/P38、P-JNK/JNK的表达较正常组增加(P<0.05);OMT可以下调TLR4、P-P38/P38、P-JNK/JNK的表达(P<0.01,P<0.05),OMT+FYK-M+LPS组及OMT+FYK-H+LPS组TLR4、P-P38/P38低于OMT组(P<0.05)。OMT+FYK-H+LPS组P-JNK/JNK低于OMT组(P<0.05)。RT-q PCR结果显示:LPS组Bax/Bcl-2及Caspase-3 m RNA表达较正常组明显增高(P<0.01)。OMT能够降低Bax/Bcl-2及Caspase-3 m RNA的水平(P<0.05)。OMT+FYK-M+LPS组、OMT+FYK-H+LPS组Bax/Bcl-2及Caspase-3 m RNA表达低于OMT组(P<0.05,P<0.01)。结论:OMT联合FYK预处理可通过TLR4/P38/JNK信号通路抑制BRL-3A细胞凋亡。展开更多
基金Postgraduate Scientific Research Innovation Key Project of Hunan Province(CX20210680)Key Scientific Research Project of Hunan Provincial Department of Education(21A0224)+1 种基金Scientific Research Project of Hunan Health Commission(202204083613)CHEN Qihua National Studio for Inheritance of TCM Famous Veteran Doctors(2022)。
基金Supported by National Natural Science Foundation of China(No.81973908)Natural Science Foundation of Heilongjiang Province(No.PL2024H224)Postgraduate Innovation Fund of Heilongjiang University of Chinese Medicine(No.2023yjscx018).
文摘AIM:To investigate the effects of a Chinese medicine formula“Qingxuan Runmu Yin”(QRY)on ocular surface inflammation in a rat model of dry eye,and its mechanism via the toll-like receptor 4(TLR4)/transforming growth factor kinase 1(TAK1)/p38 mitogen-activated protein kinase(p38MAPK)signaling pathway.METHODS:Seventy-two Sprague-Dawley rats were randomly divided into six groups(n=12 each):the control group,model group,3 groups of QRY(with low-,medium-,and high-doses),and SB203580 group.Dry eye was induced using benzalkonium chloride.Schirmer’s test(SIT)and corneal fluorescein staining(CFS)were performed every 14d throughout the experiment.Histopathological changes in corneal and conjunctival tissues were observed using hematoxylin and eosin(HE)and periodic acid-Schiff(PAS)staining.Protein expression levels of key inflammatory markers and signaling pathway targets were assessed via immunohistochemistry,ELISA,and Western blotting.RESULTS:Compared to the control group,the model group showed significant reductions in SIT and increases in CFS scores,alongside structural disorganization of corneal/conjunctival tissues,decreased conjunctival goblet cell(CGC)numbers,and elevated expression of inflammatory markers[interleukin(IL)-1β,IL-6,tumor necrosis factoralpha(TNF-α),matrix metalloproteinase-9(MMP9)]and pathway proteins(TLR4,p-TAK1,p-p38MAPK;P<0.05).Treatment with QRY(low,medium,and high doses)and SB203580 significantly improved SIT scores,reduced CFS scores,restored corneoconjunctival structure,increased CGC numbers,and decreased expression levels of IL-1β,IL-6,TNF-α,MMP9,TLR4,p-TAK1,and p-p38MAPK proteins compared to the model group(P<0.05).CONCLUSION:QRY may alleviate ocular surface inflammation associated with dry eye by inhibiting the TLR4/TAK1/p38MAPK signaling pathway,highlighting its potential therapeutic efficacy for dry eye.
文摘目的:探讨氧化苦参碱(OMT)联合复方茵陈颗粒(FYK)对脂多糖(LPS)诱导的大鼠肝BRL-3A细胞凋亡的抑制作用及其机制,为该联合用药抑制肝细胞凋亡提供更多的实验依据。方法:采用LPS诱导大鼠肝BRL-3A细胞建立细胞凋亡模型。应用CCK-8法测定BRL-3A细胞活力;流式细胞术检测细胞凋亡率;Western blot法检测BRL-3A细胞中TLR4、P38、P-P38、JNK及P-JNK蛋白的表达;RT-q PCR法检测BRL-3A细胞中Bax、Bcl-2及Caspase-3 m RNA的表达。结果:CCK-8法结果显示,OMT可以减弱由LPS引起的细胞活力降低(P<0.05),OMT+FYK-M+LPS组及OMT+FYK-H+LPS组细胞活力高于OMT组(P<0.05);OMT预处理后细胞凋亡率低于LPS组(P<0.05),OMT+FYK-M+LPS组及OMT+FYK-H+LPS组细胞凋亡率低于OMT组(P<0.05)。Western blot结果显示,LPS组TLR4、P-P38/P38、P-JNK/JNK的表达较正常组增加(P<0.05);OMT可以下调TLR4、P-P38/P38、P-JNK/JNK的表达(P<0.01,P<0.05),OMT+FYK-M+LPS组及OMT+FYK-H+LPS组TLR4、P-P38/P38低于OMT组(P<0.05)。OMT+FYK-H+LPS组P-JNK/JNK低于OMT组(P<0.05)。RT-q PCR结果显示:LPS组Bax/Bcl-2及Caspase-3 m RNA表达较正常组明显增高(P<0.01)。OMT能够降低Bax/Bcl-2及Caspase-3 m RNA的水平(P<0.05)。OMT+FYK-M+LPS组、OMT+FYK-H+LPS组Bax/Bcl-2及Caspase-3 m RNA表达低于OMT组(P<0.05,P<0.01)。结论:OMT联合FYK预处理可通过TLR4/P38/JNK信号通路抑制BRL-3A细胞凋亡。
