The Research Of Ginkgo Biloba Leaf Polysaccharides For Anti-inflammation Effect

Ginkgo biloba L belonged to Ginkoaceae is a famous plantin the world. Polysaccharides of Ginkgo biloba are a kindof functional components which have important physiologicaland pharmacological activities. Polysaccharides of Ginkgobiloba Leave(PGBL)have strong anti-inflammation effect,but its mechanism is unclear. To make it clearly, Weextracted PGBL from Ginkgo biloba leaf, established anurgent peritoneal inflammation model in mice and used flowchamber and flow cytometer in vitro to study its anti-inflammation effect and mechanism.1. PGBL inhibit urgent peritoneal inflammation in vivoThe model of urgent peritoneal inflammation in miceinduced by thioglycollate broth was established, and micewere injected with PGBL(1.25 mg), the number of neutrophilin total peritoneal cells was counted with hemocytometer.PGBL,heparin and positive group number of neutrophil is15.06 X 108/L,18.55 X 108/L and 60 X 108/L. The results showedthat heparin and PGBL groups have significant distinctioncompared with positive group, and no different between them.Then, we can see that PGBL has significant anti-inflammationeffect on urgent peritoneal inflammation in mice, and it canbe used for a new anti-inflammation drug.2 The effect of PGBL on HL-60 cell adhesion(1) HL-60 cell and CHO-P cell adhesion in flow chamberThe experiment of HL-60 adhesion to CHO-P cells in flowchamber indicated that 0.1g/L,0.2g/L and 0.5g/L dose PGBLand heparin group inhibition rate is 27.30%,31.56%,38.12% , 48.94 % respectively. It is significant differencecompared with positive control group. The result showed PGBLcan block HL-60 cell adhesion to CHO-P cell, the PGBLanti-inflammation molecular mechanism probably blocked theinteraction of P-selectin and its ligand, that is similarto heparin.(2) HL-60 cell and P-FC interaction in flow cytometreTo detect the blocking effect of PGBL and heparin, weperformed the interaction experiment of P-FC and HL-60 cellwith flow cytometrer. The result showed that PGBL(0.1g/L,0.2g/L and 0.5g/L)and heparin(1g/L) group inhibition rateis 28.66%,35.72%,38.28%and 56.97% respectively. It issignificant difference compared with positive control group.It appears that PGBL can block the interaction ofP-selectin and its ligand .3.The effect of PGBL on neutrophil adhesion(1) Neutrophil and CHO-P cell adhesion in flow chamberIn vitro, with the experiment of neutrophil adhesion toCHO-P cell in flow chamber, we investigated theanti-inflammation mechanism of PGBL. The result showed PGBLcan block neutrophil adhesion to CHO-P cell, PGBL 0.1g/L,0.2g/L and 0.5g/L dose and heparin group inhibition rate is41.26%,54.92%,60.38%and51.91% respectively,it hassignificant distinction compared with positive controlgroup. It indicated that PGBL can block neutrophil andP-selectin interaction, and participate in anti-inflammation process by blocking the ligand similar toheparin.(2) Neutrophil and P-FC interaction in flow cytometreTo detect the PGBL and heparin blocking effects, we didsome experiments on the interaction of P-FC and neutrophilby flow cytometrer. The result showed that PGBL 0.1g/L,0.2g/L and 0.5g/L dose and heparin group inhibition rateis 36.52%,40.42%,62.92% and 56.98% respectively,ithas significant distinction compared with positive controlgroup, it indicated that PGBL can inhibit the ligand bindingto neutrophil, block neutrophil adhesion to endothelial cellwhich expressed P-selectin to reach the inflammation zonethrough endothelial.Our results have demonstrated that PGBL have stronganti-inflammation effect. The anti-inflammation moleculemechanism is blocking the interaction of P-selectin and itsligand, and then inhibiting neutrophil adhesion to vascularendothelial cell. Our data provides fundamental research forexploiting PGBL as a new anti-inflammation drug.