目的:明确P38/丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)通路在脉冲式Nd:YAG激光促进口腔溃疡愈合中的作用。方法:SD雄性大鼠共70只,随机分为正常组、溃疡组、自愈组、封闭组、激光组、抑制剂组和激动剂组,每组各10...目的:明确P38/丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)通路在脉冲式Nd:YAG激光促进口腔溃疡愈合中的作用。方法:SD雄性大鼠共70只,随机分为正常组、溃疡组、自愈组、封闭组、激光组、抑制剂组和激动剂组,每组各10只。除正常组直接切取下唇口腔黏膜组织外,其余组别均采用创伤法建立口腔溃疡模型。溃疡组在溃疡形成当日(第2天)取材,其余5组经不同处理后于第4天取材。观察大鼠的创面愈合情况;利用蛋白质印迹分析检测口腔黏膜组织中P38、p-P38的表达;免疫组织化学技术检测白细胞介素17(interleukin 17,IL-17)、基质金属蛋白酶(Matrix metalloproteinase,MMPs)-2、MMP-9、基质金属蛋白酶组织抑制剂(tissue inhibitor of metalloproteinase,TIMPs)-1以及Ⅳ型胶原蛋白的表达。结果:大鼠的创面愈合情况:封闭组、激光组和抑制剂组大鼠口腔溃疡面积明显缩小;自愈组和激动剂组大鼠溃疡表面仍存在假膜和脓液。Western-blot结果:与正常组相比,溃疡组大鼠中p-P38的表达升高(P<0.05)。与溃疡组相比,封闭组和激光组大鼠中p-P38的表达降低(P<0.05);自愈组中p-P38的表达虽有所降低,但差异无统计学意义(P>0.05);与激光组相比,激动剂组中p-P38的表达显著升高(P<0.01);抑制剂组中p-P38的表达虽有所降低,但差异无统计学意义(P>0.05)。P38总蛋白在各组中的表达均未见明显差异(P>0.05)。免疫组化结果:与正常组相比,溃疡组大鼠组织中IL-17、MMP-2和MMP-9的表达升高(P<0.05;P<0.001;P<0.0001),TIMP-1和Ⅳ型胶原蛋白的表达显著降低(P<0.001;P<0.0001;)。与溃疡组相比,封闭组和激光组大鼠中IL-17(P<0.01;P<0.001)、MMP-2(P<0.01)和MMP-9(P<0.001;P<0.0001)的表达显著降低,TIMP-1(P<0.01)和Ⅳ型胶原蛋白(P<0.0001)的表达显著升高;与自愈组相比,封闭组和激光组大鼠中IL-17的表达降低(P>0.05;P<0.05),但封闭组差异无统计学意义;封闭组和激光组大鼠中MMP-2(P<0.05)和MMP-9(P<0.05;P<0.001)的表达降低;TIMP-1(P>0.05;P<0.01)的表达升高,但封闭组差异无统计学意义;Ⅳ型胶原蛋白(P<0.0001)的表达显著升高;与激光组相比,激动剂组大鼠中IL-17、MMP-2和MMP-9的表达显著升高(P<0.001),TIMP-1(P<0.05)和Ⅳ型胶原蛋白(P<0.01)的表达降低。结论:脉冲式Nd:YAG激光可以通过抑制P38/MAPK信号通路,缩短口腔溃疡的愈合时间,达到促进溃疡组织愈合的效果;IL-17可以通过P38/MAPK信号通路调节MMPs/TIMPs表达平衡,促进Ⅳ型胶原蛋白的合成,从而在脉冲式Nd:YAG激光促进口腔溃疡愈合的过程中发挥作用。展开更多
Background:Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-related mortality worldwide.This study aimed to identify key genes involved in HCC development and elucidate their molecular mechanisms,wi...Background:Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-related mortality worldwide.This study aimed to identify key genes involved in HCC development and elucidate their molecular mechanisms,with a particular focus on mitochondrial function and apoptosis.Methods:Differential expression analyses were performed across three datasets—The Cancer Genome Atlas(TCGA)-Liver Hepatocellular Carcinoma(LIHC),GSE36076,and GSE95698—to identify overlapping differentially expressed genes(DEGs).A prognostic risk model was then constructed.Cysteine/serine-rich nuclear protein 1(CSRNP1)expression levels in HCC cell lines were assessed via western blot(WB)and quantitative reverse transcription polymerase chain reaction(qRT-PCR).The effects of CSRNP1 knockdown or overexpression on cell proliferation,migration,and apoptosis were evaluated using cell counting-8(CCK-8)assays,Transwell assays,and flow cytometry.Mitochondrial ultrastructure was examined by transmission electron microscopy,and intracellular and mitochondrial reactive oxygen species(mROS)levels were measured using specific fluorescent probes.WB was used to assess activation of the c-Jun N-terminal kinase(JNK)/p38 mitogen-activated protein kinase(MAPK)pathway,and pathway dependence was examined using the ROS scavenger N-Acetylcysteine(NAC)and the JNK inhibitor SP600125.Results:A six-gene prognostic model was established,comprising downregulated genes(NR4A1 and CSRNP1)and upregulated genes(CENPQ,YAE1,FANCF,and POC5)in HCC.Functional experiments revealed that CSRNP1 knockdown promoted the proliferation of HCC cells and suppressed their apoptosis.Conversely,CSRNP1 overexpression impaired mitochondrial integrity,increased both mitochondrial and cytoplasmic ROS levels,and activated the JNK/p38 MAPK pathway.Notably,treatment with NAC or SP600125 attenuated CSRNP1-induced MAPK activation and apoptosis.Conclusion:CSRNP1 is a novel prognostic biomarker and tumor suppressor in HCC.It exerts anti-tumor effects by inducing oxidative stress and activating the JNK/p38 MAPK pathway in a ROS-dependent manner.These findings suggest that CSRNP1 may serve as a potential therapeutic target in the management of HCC.展开更多
文摘目的:明确P38/丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)通路在脉冲式Nd:YAG激光促进口腔溃疡愈合中的作用。方法:SD雄性大鼠共70只,随机分为正常组、溃疡组、自愈组、封闭组、激光组、抑制剂组和激动剂组,每组各10只。除正常组直接切取下唇口腔黏膜组织外,其余组别均采用创伤法建立口腔溃疡模型。溃疡组在溃疡形成当日(第2天)取材,其余5组经不同处理后于第4天取材。观察大鼠的创面愈合情况;利用蛋白质印迹分析检测口腔黏膜组织中P38、p-P38的表达;免疫组织化学技术检测白细胞介素17(interleukin 17,IL-17)、基质金属蛋白酶(Matrix metalloproteinase,MMPs)-2、MMP-9、基质金属蛋白酶组织抑制剂(tissue inhibitor of metalloproteinase,TIMPs)-1以及Ⅳ型胶原蛋白的表达。结果:大鼠的创面愈合情况:封闭组、激光组和抑制剂组大鼠口腔溃疡面积明显缩小;自愈组和激动剂组大鼠溃疡表面仍存在假膜和脓液。Western-blot结果:与正常组相比,溃疡组大鼠中p-P38的表达升高(P<0.05)。与溃疡组相比,封闭组和激光组大鼠中p-P38的表达降低(P<0.05);自愈组中p-P38的表达虽有所降低,但差异无统计学意义(P>0.05);与激光组相比,激动剂组中p-P38的表达显著升高(P<0.01);抑制剂组中p-P38的表达虽有所降低,但差异无统计学意义(P>0.05)。P38总蛋白在各组中的表达均未见明显差异(P>0.05)。免疫组化结果:与正常组相比,溃疡组大鼠组织中IL-17、MMP-2和MMP-9的表达升高(P<0.05;P<0.001;P<0.0001),TIMP-1和Ⅳ型胶原蛋白的表达显著降低(P<0.001;P<0.0001;)。与溃疡组相比,封闭组和激光组大鼠中IL-17(P<0.01;P<0.001)、MMP-2(P<0.01)和MMP-9(P<0.001;P<0.0001)的表达显著降低,TIMP-1(P<0.01)和Ⅳ型胶原蛋白(P<0.0001)的表达显著升高;与自愈组相比,封闭组和激光组大鼠中IL-17的表达降低(P>0.05;P<0.05),但封闭组差异无统计学意义;封闭组和激光组大鼠中MMP-2(P<0.05)和MMP-9(P<0.05;P<0.001)的表达降低;TIMP-1(P>0.05;P<0.01)的表达升高,但封闭组差异无统计学意义;Ⅳ型胶原蛋白(P<0.0001)的表达显著升高;与激光组相比,激动剂组大鼠中IL-17、MMP-2和MMP-9的表达显著升高(P<0.001),TIMP-1(P<0.05)和Ⅳ型胶原蛋白(P<0.01)的表达降低。结论:脉冲式Nd:YAG激光可以通过抑制P38/MAPK信号通路,缩短口腔溃疡的愈合时间,达到促进溃疡组织愈合的效果;IL-17可以通过P38/MAPK信号通路调节MMPs/TIMPs表达平衡,促进Ⅳ型胶原蛋白的合成,从而在脉冲式Nd:YAG激光促进口腔溃疡愈合的过程中发挥作用。
基金funded by Shanghai Yangpu District Science and Technology Commission(Grant No.YPQ202303(Xuejing Lin))Shanghai Yangpu Hospital Foundation(Grant No.Se1202420(Wenchao Wang)and Ye1202423(Juan Huang)).
文摘Background:Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-related mortality worldwide.This study aimed to identify key genes involved in HCC development and elucidate their molecular mechanisms,with a particular focus on mitochondrial function and apoptosis.Methods:Differential expression analyses were performed across three datasets—The Cancer Genome Atlas(TCGA)-Liver Hepatocellular Carcinoma(LIHC),GSE36076,and GSE95698—to identify overlapping differentially expressed genes(DEGs).A prognostic risk model was then constructed.Cysteine/serine-rich nuclear protein 1(CSRNP1)expression levels in HCC cell lines were assessed via western blot(WB)and quantitative reverse transcription polymerase chain reaction(qRT-PCR).The effects of CSRNP1 knockdown or overexpression on cell proliferation,migration,and apoptosis were evaluated using cell counting-8(CCK-8)assays,Transwell assays,and flow cytometry.Mitochondrial ultrastructure was examined by transmission electron microscopy,and intracellular and mitochondrial reactive oxygen species(mROS)levels were measured using specific fluorescent probes.WB was used to assess activation of the c-Jun N-terminal kinase(JNK)/p38 mitogen-activated protein kinase(MAPK)pathway,and pathway dependence was examined using the ROS scavenger N-Acetylcysteine(NAC)and the JNK inhibitor SP600125.Results:A six-gene prognostic model was established,comprising downregulated genes(NR4A1 and CSRNP1)and upregulated genes(CENPQ,YAE1,FANCF,and POC5)in HCC.Functional experiments revealed that CSRNP1 knockdown promoted the proliferation of HCC cells and suppressed their apoptosis.Conversely,CSRNP1 overexpression impaired mitochondrial integrity,increased both mitochondrial and cytoplasmic ROS levels,and activated the JNK/p38 MAPK pathway.Notably,treatment with NAC or SP600125 attenuated CSRNP1-induced MAPK activation and apoptosis.Conclusion:CSRNP1 is a novel prognostic biomarker and tumor suppressor in HCC.It exerts anti-tumor effects by inducing oxidative stress and activating the JNK/p38 MAPK pathway in a ROS-dependent manner.These findings suggest that CSRNP1 may serve as a potential therapeutic target in the management of HCC.
