Traumatic brain injury is a severe health problem leading to autophagy and apoptosis in the brain.3,6-Dibromo-beta-fluoro-N-(3-methoxyphenyl)-9H-carbazole-9-propanamine(P7C3-A20)can be neuroprotective in various disea...Traumatic brain injury is a severe health problem leading to autophagy and apoptosis in the brain.3,6-Dibromo-beta-fluoro-N-(3-methoxyphenyl)-9H-carbazole-9-propanamine(P7C3-A20)can be neuroprotective in various diseases,including ischemic stroke and neurodegenerative diseases.However,whether P7C3-A20 has a therapeutic effect on traumatic brain injury and its possible molecular mechanisms are unclear.Therefore,in the present study,we investigated the therapeutic effects of P7C3-A20 on traumatic brain injury and explored the putative underlying molecular mechanisms.We established a traumatic brain injury rat model using a modified weight drop method.P7C3-A20 or vehicle was injected intraperitoneally after traumatic brain injury.Severe neurological deficits were found in rats after traumatic brain injury,with deterioration in balance,walking function,and learning memory.Furthermore,hematoxylin and eosin staining showed significant neuronal cell damage,while terminal deoxynucleotidyl transferase mediated dUTP nick end labeling staining indicated a high rate of apoptosis.The presence of autolysosomes was observed using transmission electron microscope.P7C3-A20 treatment reversed these pathological features.Western blotting showed that P7C3-A20 treatment reduced microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)autophagy protein,apoptosis-related proteins(namely,Bcl-2/adenovirus E1B 19-kDa-interacting protein 3[BNIP3],and Bcl-2 associated x protein[Bax]),and elevated ubiquitin-binding protein p62(p62)autophagy protein expression.Thus,P7C3-A20 can treat traumatic brain injury in rats by inhibiting excessive autophagy and apoptosis.展开更多
目的:探讨miR-20a-3p基因干扰对人牙髓干细胞(hDPSCs)成牙本质分化的影响及信号通路调控机制。方法:对hDPSCs转染miR-20a-3p-mimic和miR-20a-3p-inhibitor来上调或下调其表达,然后对hDPSCs进行成牙本质分化诱导培养。通过茜素红染色观...目的:探讨miR-20a-3p基因干扰对人牙髓干细胞(hDPSCs)成牙本质分化的影响及信号通路调控机制。方法:对hDPSCs转染miR-20a-3p-mimic和miR-20a-3p-inhibitor来上调或下调其表达,然后对hDPSCs进行成牙本质分化诱导培养。通过茜素红染色观察细胞钙基质的矿化程度并检测ALP活性。通过双荧光素酶报告基因测定实验验证miR-20a-3p与mothers against decapentaplegic homolog(SMAD)特异性E3泛素蛋白连接酶1(SMURF1)的靶向调节关系。通过RT-qPCR检测hDPSCs中miR-20a-3p以及SMURF1、Runt相关转录因子2(RUNX2)、骨钙蛋白(OCN)和牙本质涎磷蛋白(DSPP)mRNA水平。结果:与Control组相比,miR-20a-3p-mimic组的茜素红相对染色强度升高了67.09%,miR-20a-3p-inhibitor组降低了46.13%(P<0.05)。与Control组相比,miR-20a-3p-mimic组的相对ALP活性升高了52.89%,miR-20a-3p-inhibitor组降低了53.32%(P<0.05)。与Control组相比,miR-20a-3p-mimic组的RUNX2、OCN和DSPP mRNA相对表达量分别升高了2.19倍、1.86倍和2.35倍,miR-20a-3p-inhibitor组分别降低了63.26%、58.84%和68.12%(P<0.05)。双荧光素酶报告基因测定显示miR-20a-3p靶向抑制SMURF1(P<0.05)。过表达SMURF1抑制了miR-20a-3p对成牙本质分化的影响(P<0.05)。结论:miR-20a-3p通过靶向抑制SMURF1及其下游基因促进hDPSCs的成牙本质分化。展开更多
基金supported by National Natural Science Foundation of China,No.32102745(to XL).
文摘Traumatic brain injury is a severe health problem leading to autophagy and apoptosis in the brain.3,6-Dibromo-beta-fluoro-N-(3-methoxyphenyl)-9H-carbazole-9-propanamine(P7C3-A20)can be neuroprotective in various diseases,including ischemic stroke and neurodegenerative diseases.However,whether P7C3-A20 has a therapeutic effect on traumatic brain injury and its possible molecular mechanisms are unclear.Therefore,in the present study,we investigated the therapeutic effects of P7C3-A20 on traumatic brain injury and explored the putative underlying molecular mechanisms.We established a traumatic brain injury rat model using a modified weight drop method.P7C3-A20 or vehicle was injected intraperitoneally after traumatic brain injury.Severe neurological deficits were found in rats after traumatic brain injury,with deterioration in balance,walking function,and learning memory.Furthermore,hematoxylin and eosin staining showed significant neuronal cell damage,while terminal deoxynucleotidyl transferase mediated dUTP nick end labeling staining indicated a high rate of apoptosis.The presence of autolysosomes was observed using transmission electron microscope.P7C3-A20 treatment reversed these pathological features.Western blotting showed that P7C3-A20 treatment reduced microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)autophagy protein,apoptosis-related proteins(namely,Bcl-2/adenovirus E1B 19-kDa-interacting protein 3[BNIP3],and Bcl-2 associated x protein[Bax]),and elevated ubiquitin-binding protein p62(p62)autophagy protein expression.Thus,P7C3-A20 can treat traumatic brain injury in rats by inhibiting excessive autophagy and apoptosis.
文摘目的:探讨miR-20a-3p基因干扰对人牙髓干细胞(hDPSCs)成牙本质分化的影响及信号通路调控机制。方法:对hDPSCs转染miR-20a-3p-mimic和miR-20a-3p-inhibitor来上调或下调其表达,然后对hDPSCs进行成牙本质分化诱导培养。通过茜素红染色观察细胞钙基质的矿化程度并检测ALP活性。通过双荧光素酶报告基因测定实验验证miR-20a-3p与mothers against decapentaplegic homolog(SMAD)特异性E3泛素蛋白连接酶1(SMURF1)的靶向调节关系。通过RT-qPCR检测hDPSCs中miR-20a-3p以及SMURF1、Runt相关转录因子2(RUNX2)、骨钙蛋白(OCN)和牙本质涎磷蛋白(DSPP)mRNA水平。结果:与Control组相比,miR-20a-3p-mimic组的茜素红相对染色强度升高了67.09%,miR-20a-3p-inhibitor组降低了46.13%(P<0.05)。与Control组相比,miR-20a-3p-mimic组的相对ALP活性升高了52.89%,miR-20a-3p-inhibitor组降低了53.32%(P<0.05)。与Control组相比,miR-20a-3p-mimic组的RUNX2、OCN和DSPP mRNA相对表达量分别升高了2.19倍、1.86倍和2.35倍,miR-20a-3p-inhibitor组分别降低了63.26%、58.84%和68.12%(P<0.05)。双荧光素酶报告基因测定显示miR-20a-3p靶向抑制SMURF1(P<0.05)。过表达SMURF1抑制了miR-20a-3p对成牙本质分化的影响(P<0.05)。结论:miR-20a-3p通过靶向抑制SMURF1及其下游基因促进hDPSCs的成牙本质分化。
