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
Children with autism often exhibit abnormalities in body weight,but the underlying mechanism remains unclear.SH3 and multiple ankyrin repeat domains protein 3(SHANK3),a scaffold protein of the postsynaptic density,has...Children with autism often exhibit abnormalities in body weight,but the underlying mechanism remains unclear.SH3 and multiple ankyrin repeat domains protein 3(SHANK3),a scaffold protein of the postsynaptic density,has been reported to be associated with autism.This study aimed to investigate whether and how SHANK3 influences body weight in the hypothalamic neuronal regulation of energy homeostasis.Adeno-associated viruses 9(AAV9)carrying CMV-Cre and Agrp-Cre were stereotactically injected to restore SHANK3 expression in the arcuate nucleus(ARC)and agouti-related peptide(AgRP)neurons,respectively.Agrp-Cre mice were injected with AAV9-p38αflox/flox to overexpress p38α.Activated p38αwas generated by mutating both D176A and F327S in p38α.Inactivated p38αwas constructed by mutating both T180A and Y182F in p38α.Metabolic analysis,immunoblotting,histological analysis,the glucose tolerance test,the insulin tolerance test,and body fat mass analysis were applied to investigate the underlying mechanisms by which SHANK3 regulates body weight.We reveal that SHANK3 regulates body weight via the p38αsignaling pathway in the AgRP neurons of the hypothalamus.Shank3 knockout(Shank3−/−)mice exhibit resistance to diet-induced obesity.Shank3 re-expression in the ARC or AgRP neurons increases body weight in Shank3 knock-in mice with an inverted allele(SKO).Overexpression or activation of p38αin AgRP neurons elicits resistance to diet-induced obesity.Inactivated p38αin AgRP neurons abolished the resistance to diet-induced obesity due to SHANK3 deficiency.Our findings suggest that the SHANK3-p38αsiganling pathway in AgRP neurons regulates body weight balance in autism,revealing a promising therapeutic target for obesity in children with autism.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supported by the Noncommunicable Chronic Diseases-National Science and Technology Major Project(2023ZD0506800).
文摘Children with autism often exhibit abnormalities in body weight,but the underlying mechanism remains unclear.SH3 and multiple ankyrin repeat domains protein 3(SHANK3),a scaffold protein of the postsynaptic density,has been reported to be associated with autism.This study aimed to investigate whether and how SHANK3 influences body weight in the hypothalamic neuronal regulation of energy homeostasis.Adeno-associated viruses 9(AAV9)carrying CMV-Cre and Agrp-Cre were stereotactically injected to restore SHANK3 expression in the arcuate nucleus(ARC)and agouti-related peptide(AgRP)neurons,respectively.Agrp-Cre mice were injected with AAV9-p38αflox/flox to overexpress p38α.Activated p38αwas generated by mutating both D176A and F327S in p38α.Inactivated p38αwas constructed by mutating both T180A and Y182F in p38α.Metabolic analysis,immunoblotting,histological analysis,the glucose tolerance test,the insulin tolerance test,and body fat mass analysis were applied to investigate the underlying mechanisms by which SHANK3 regulates body weight.We reveal that SHANK3 regulates body weight via the p38αsignaling pathway in the AgRP neurons of the hypothalamus.Shank3 knockout(Shank3−/−)mice exhibit resistance to diet-induced obesity.Shank3 re-expression in the ARC or AgRP neurons increases body weight in Shank3 knock-in mice with an inverted allele(SKO).Overexpression or activation of p38αin AgRP neurons elicits resistance to diet-induced obesity.Inactivated p38αin AgRP neurons abolished the resistance to diet-induced obesity due to SHANK3 deficiency.Our findings suggest that the SHANK3-p38αsiganling pathway in AgRP neurons regulates body weight balance in autism,revealing a promising therapeutic target for obesity in children with autism.
