已报道低浓度溶血磷脂酸(lysophosphatidic acid,LPA)对去血清培养所致的神经元凋亡有神经保护作用.为了进一步观察LPA是否对β-amyloid peptide fragment 31-35(AβP31-35)所致的神经元凋亡也起类似的作用,本研究应用DNA电泳分析、HO33...已报道低浓度溶血磷脂酸(lysophosphatidic acid,LPA)对去血清培养所致的神经元凋亡有神经保护作用.为了进一步观察LPA是否对β-amyloid peptide fragment 31-35(AβP31-35)所致的神经元凋亡也起类似的作用,本研究应用DNA电泳分析、HO33342和TUNEL染色法等技术,对培养的小鼠大脑皮层神经元进行了观察.结果显示,只有使用较低浓度的LPA(1～10μmol/L)、并且将此剂量的LPA比AβP31-35提前12～24 h加入培养液时,才可看到LPA明显削弱了AβP31-35所致的神经元凋亡.以上结果表明,适当浓度的LPA在长时间预作用的条件下,可对AβP31-35所致的皮层神经元凋亡起保护因子或抗凋亡因子的作用,但其作用途径可能较在去血清培养所致的凋亡时更为复杂,因为在去血清的同时加入LPA就能制止去血清所致的凋亡.展开更多
Background We previously reported that iodine-131（131^I）-Iabeled anti-pro-gastrin-releasing peptide （ProGRP（31-98）） monoclonal antibody D-D3 could selectively accumulate in the tumor sites of nude mice bearing s...Background We previously reported that iodine-131（131^I）-Iabeled anti-pro-gastrin-releasing peptide （ProGRP（31-98）） monoclonal antibody D-D3 could selectively accumulate in the tumor sites of nude mice bearing small cell lung cancer （SCLC） xenografts. However, 1311-D-D3 was cleared slowly from the body, and the best radioimmunoimaging time for SCLC was 72-96 hours after injection. The aims of this study were to radiolabel anti-ProGRP（31-98） D-D3 monoclonal antibody with technetium-99m（99m^Tc） and to investigate the biodistribution of this antibody in healthy ICR mice. Methods D-D3was labeled with 99m^Tc via the 2-mercaptoethanol reduction method. 99m^Tc-D-g3 was purified by the gel column separation method. The labeling efficiency and radiochemical purity were measured by thin-layer chromatography. The immunological activity of 99m^Tc-D-O3 was determined with cell conjugation assays. 99m^Tc-D-D3 was injected into healthy ICR mice via a tail vein, and all the healthy ICR mice were sacrificed by cervical dislocation at a designated time. Then, the blood and major organs were removed and weighed, and counted in a gamma scintillation counter to determine the percentage of the injected dose per gram （%ID/g）. Results The labeling rate and the radiochemical purity of 99m^TC-D-D3 were （73.87±2.89）% and （94.13±4.49）%, respectively. The immunobinding rates of 99m^Tc-O-D3 to the human small cell lung cancer NCI-H446 cell line and lung adenocarcinoma A549 cell line were （81.2±2.37）% and （24.3±1.46）%, respectively. The distribution data of normal ICR mice demonstrated that 99m^TC-D-D3was mainly distributed in the liver, kidney and lung, and less in the brain tissue and muscle. Conclusions 99m^Tc-D-D3 antibody not only had high radiochemical purity, but also had good stability both in vitro and in vivo, and maintained good immunological activity. 99m^Tc-D-D3 was metabolized mainly in the kidney and liver, and the blood radioactivity decreased rapidly. Thus, 99m^Tc-O-D3 is conducive to the radioimmunoimaging of SCLC.展开更多
Mitochondrial dysfunction is associated with the emergence of several neurological and cardiovascular diseases. Hence, mitochondria-targeting delivery strategies are highly significant and critically needed. In this s...Mitochondrial dysfunction is associated with the emergence of several neurological and cardiovascular diseases. Hence, mitochondria-targeting delivery strategies are highly significant and critically needed. In this study, we developed a small library of peptides simulating the mitochondria-targeting peptide SS-31, a promising tetra-peptide with antioxidant character, and subsequently evaluated the toxicity, antioxidant ability and mitochondrial delivery of nanoparticles. Among the designed peptides, RF-2 (DArg-Dmt-Arg-Phe-NH2) showed controlled toxicity and excellent protection against gentamicin-induced hair cell damage, as compared with SS-31. More importantly, RF-2-modified PLGA nanoparticles demonstrated high colocalization with mitochondria and comparable specific subcellular accumulation, when compared with SS-31. Taken together, the obtained results supported RF-2 as a mitochondriatargeting peptide with high potential as a targeted carrier.展开更多
文摘Background We previously reported that iodine-131（131^I）-Iabeled anti-pro-gastrin-releasing peptide （ProGRP（31-98）） monoclonal antibody D-D3 could selectively accumulate in the tumor sites of nude mice bearing small cell lung cancer （SCLC） xenografts. However, 1311-D-D3 was cleared slowly from the body, and the best radioimmunoimaging time for SCLC was 72-96 hours after injection. The aims of this study were to radiolabel anti-ProGRP（31-98） D-D3 monoclonal antibody with technetium-99m（99m^Tc） and to investigate the biodistribution of this antibody in healthy ICR mice. Methods D-D3was labeled with 99m^Tc via the 2-mercaptoethanol reduction method. 99m^Tc-D-g3 was purified by the gel column separation method. The labeling efficiency and radiochemical purity were measured by thin-layer chromatography. The immunological activity of 99m^Tc-D-O3 was determined with cell conjugation assays. 99m^Tc-D-D3 was injected into healthy ICR mice via a tail vein, and all the healthy ICR mice were sacrificed by cervical dislocation at a designated time. Then, the blood and major organs were removed and weighed, and counted in a gamma scintillation counter to determine the percentage of the injected dose per gram （%ID/g）. Results The labeling rate and the radiochemical purity of 99m^TC-D-D3 were （73.87±2.89）% and （94.13±4.49）%, respectively. The immunobinding rates of 99m^Tc-O-D3 to the human small cell lung cancer NCI-H446 cell line and lung adenocarcinoma A549 cell line were （81.2±2.37）% and （24.3±1.46）%, respectively. The distribution data of normal ICR mice demonstrated that 99m^TC-D-D3was mainly distributed in the liver, kidney and lung, and less in the brain tissue and muscle. Conclusions 99m^Tc-D-D3 antibody not only had high radiochemical purity, but also had good stability both in vitro and in vivo, and maintained good immunological activity. 99m^Tc-D-D3 was metabolized mainly in the kidney and liver, and the blood radioactivity decreased rapidly. Thus, 99m^Tc-O-D3 is conducive to the radioimmunoimaging of SCLC.
基金financially supported by the National Natural Science Foundation of China (No. 81473162)the Program for Liaoning Excellent Talents in Universitythe Young and Middleaged Career Development Planning of Shenyang Pharmaceutical University
文摘Mitochondrial dysfunction is associated with the emergence of several neurological and cardiovascular diseases. Hence, mitochondria-targeting delivery strategies are highly significant and critically needed. In this study, we developed a small library of peptides simulating the mitochondria-targeting peptide SS-31, a promising tetra-peptide with antioxidant character, and subsequently evaluated the toxicity, antioxidant ability and mitochondrial delivery of nanoparticles. Among the designed peptides, RF-2 (DArg-Dmt-Arg-Phe-NH2) showed controlled toxicity and excellent protection against gentamicin-induced hair cell damage, as compared with SS-31. More importantly, RF-2-modified PLGA nanoparticles demonstrated high colocalization with mitochondria and comparable specific subcellular accumulation, when compared with SS-31. Taken together, the obtained results supported RF-2 as a mitochondriatargeting peptide with high potential as a targeted carrier.
