Lv Li, Lu Qian, Shao Fangyu, Li Weiping, Zhou Qin, Wang Jihong, Li Qingwei
School of Life Sciences, Liaoning Normal University, Dalian, Liaoning 116029, China; Department of Pharmacology, Dalian Medical University, Dalian, Liaoning 116044, China.
Department of Pharmacology, Dalian Medical University, Dalian, Liaoning 116044, China; College of Basic Medicine, Jilin Medical University, Jilin, Jilin 132013, China.
Biomed Res Int. 2016;2016:6701249. doi: 10.1155/2016/6701249. Epub 2016 Sep 5.
rLj-RGD3 is a 14.5 kDa recombinant protein with 3 RGD (Arg-Gly-Asp) motifs from the salivary gland secretions of , which is a histidine-rich and arginine-rich protein. Previous reports indicated that rLj-RGD3 has typical functions of RGD-toxin protein, such as platelet aggregation suppression tumour metastasis and angiogenesis inhibition. Because histidine and arginine have cerebral ischemia-reperfusion and neuroprotective functions, we investigated whether rLj-RGD3 has such activities and studied the mechanism. The effects of rLj-RGD3 on neuroprotection and antiapoptosis were determined. The expression level of focal adhesion kinase (FAK), p-FAK, Caspase-3, and Bcl-2 after oxygen-glucose deprivation and reperfusion (OGD-R) was examined. The viability of PC12 cells incubated with rLj-RGD3 at high concentrations (16 mol/L) increased significantly due to its ability to protect the cells from apoptosis after OGD-R-induced injury. Furthermore, rLj-RGD3 attenuated the damage due to OGD-R. Most of the PC12 cells were apoptotic after OGD-R. In contrast, the number of apoptotic PC12 cells was significantly decreased in the group treated with a high-dose of rLj-RGD3. In addition, rLj-RGD3 activated FAK and p-FAK protein. rLj-RGD3 inhibited Caspase-3 and upregulated Bcl-2 protein expression in PC12 cells after OGD-R. The study provides the first evidence for neuroprotective effects of rLj-RGD3 in ischemic injury that may be partly mediated through inhibition of Caspase-3 and upregulation of Bcl-2, FAK, and p-FAK protein expression.
rLj-RGD3是一种14.5 kDa的重组蛋白,具有来自唾液腺分泌物的3个RGD(精氨酸-甘氨酸-天冬氨酸)基序,是一种富含组氨酸和精氨酸的蛋白质。先前的报道表明,rLj-RGD3具有RGD毒素蛋白的典型功能,如抑制血小板聚集、肿瘤转移和血管生成。由于组氨酸和精氨酸具有脑缺血再灌注和神经保护功能,我们研究了rLj-RGD3是否具有此类活性并探讨了其机制。确定了rLj-RGD3对神经保护和抗凋亡的作用。检测了氧糖剥夺和再灌注(OGD-R)后粘着斑激酶(FAK)、磷酸化FAK(p-FAK)、半胱天冬酶-3(Caspase-3)和Bcl-2的表达水平。高浓度(16 μmol/L)的rLj-RGD3孵育PC12细胞后,其活力显著增加,因为它能够保护细胞免受OGD-R诱导的损伤后的凋亡。此外,rLj-RGD3减轻了OGD-R造成的损伤。OGD-R后大多数PC12细胞发生凋亡。相比之下,高剂量rLj-RGD3处理组的凋亡PC12细胞数量显著减少。此外,rLj-RGD3激活了FAK和p-FAK蛋白。OGD-R后,rLj-RGD3抑制PC12细胞中的Caspase-3并上调Bcl-2蛋白表达。该研究首次提供了rLj-RGD3在缺血性损伤中具有神经保护作用的证据,其机制可能部分是通过抑制Caspase-3以及上调Bcl-2、FAK和p-FAK蛋白表达来介导的。