Tianjin Marine Environmental Protection and Restoration Technology Engineering Center; Tianjin Key Laboratory of Marine Resource and Chemistry; College of Marine & Environment, Tianjin University of Science and Technology, Tianjin, China.
Pak J Pharm Sci. 2021 Jan;34(1):129-134.
The aim of this study was to examine the effects of glycosaminoglycan (GAG) from Urechis unicinctus on the P2Y1 receptor pathway and expression of related factors in rat platelets. The concentration of calcium ion (Ca) in rat platelets was determined by double wavelength Fura-2 fluorescence spectrophotometry, and the concentrations of inositol trisphosphate (IP) and glycoprotein IIb/IIIa (GPIIb/IIIa) in rat platelets were measured using the enzymatic immunoassay method. The phosphorylation levels of phospholipase C (PLC), phospholipase A (PLA), protein kinase C (PKC), and p38 mitogen-activated protein kinase (p38MAPK) were also detected by Western blot. It was found that the GAG from U. unicinctus significantly reduced the Ca and IP3 levels in rat platelets (p<0.05, p<0.01). Moreover, medium and high concentrations of GAG significantly reduced the concentration of the platelet membrane GPIIb/IIIa in rats (p<0.05, p<0.01). The phosphorylation levels of PLC, PLA), PKC and p38MAPK in rat platelets were also inhibited by GAG and P)Y) receptor blocker MRS2179 (p<0.05, p<0.01). However, the degree of inhibition of GAG was lower than that of MRS2179. The results laid a foundation for further utilization of the glycosaminoglycan.
本研究旨在探讨海胆糖胺聚糖（GAG）对大鼠血小板 P2Y1 受体途径及相关因子表达的影响。采用双波长 Fura-2 荧光分光光度法测定大鼠血小板内钙离子（Ca）浓度，酶联免疫吸附法测定大鼠血小板中三磷酸肌醇（IP）和糖蛋白 IIb/IIIa（GPIIb/IIIa）的浓度。通过 Western blot 检测磷脂酶 C（PLC）、磷脂酶 A（PLA）、蛋白激酶 C（PKC）和 p38 丝裂原活化蛋白激酶（p38MAPK）的磷酸化水平。结果表明，海胆糖胺聚糖可显著降低大鼠血小板内 Ca 和 IP3 水平（p<0.05，p<0.01）。此外，中、高浓度的 GAG 可显著降低大鼠血小板膜 GPIIb/IIIa 浓度（p<0.05，p<0.01）。GAG 和 P2Y1 受体阻滞剂 MRS2179 还可抑制大鼠血小板 PLC、PLA）、PKC 和 p38MAPK 的磷酸化水平（p<0.05，p<0.01）。但 GAG 的抑制程度低于 MRS2179。该研究为进一步利用海胆糖胺聚糖奠定了基础。
