Research On The Purification Process Of Ginkgolic Acid And Its Anti-inflammatory Activity

This thesis mainly studied the purification methods of the ginkgolic acid extracts.It also studied the anti-inflammatory activity of Ginkgolic acid according to several animal models of inflammatory and the anti-inflammatory action mechanism on allergic contact dermatitis (ACD) at the level of molecular. The main contents and results are put as follows:Crude extract of ginkgolic was purified by water washing method combine with silica gel chromatography and NaOH extraction method, following detected by HPLC, then we obtained the optimal process. the purity of ginkgolic acids alcohol can be improved from 33.9% to 47.82% by washing method in solid-liquid ratio of 1:8, temperature of 65℃, washing time of 4 h; the purity of ginkgolic acids alcohol can be improved to 67.5% by Silica gel chromatography which was at 1 mL/min flow rate inject with the original concentration of ginkgolic acids, in 1 mL/min flow rate to elute after ginkgolic acids was dealed with water washing method; the purity of ginkgolic acids can be improved from 33.9% to 62.3% by NaOH solution extraction.The ginkgolic acid skin topical anti-inflammatory effects was evaluated by using xylene-induced mouse ear edema model, carrageenan-induced mouse paw edema model, allergic contact dermatitis model and the skin capillary permeability test. The results showed that Ginkgolic acids could not only inhibit ear swelling induced by xylene but also inhibit paw edema induced by carrageenan, and both ear swelling rate and paws swelling rate of the ginkgolic acids group was significantly lower than those of the model group. Moreover, ginkgolic acids could significantly inhibit mice abdominal skin capillary permeability induced by xylene. In the allergic contact dermatitis(ACD) experiment, the swelling in mice ear of the ginkgolic acids group was reduced, and the skin lesions of the ginkgolic acids group has been improved which showed that ginkgolic acids also had a good therapeutic effect on allergic contact dermatitis on mice. Histopathological results showed that ginkgolic acids could not only reduce local inflammatory cell infiltration, cellular edema, vascular dilatation in the experiment of xylene-induced ear edema; but also reduce ear inflammatory cell infiltration in these experiments of carrageenan-induced paw edema and allergic contact dermatitis. In conclusion, ginkgolic acids act a significant role in the anti-inflammation effect by inhibitting skin inflammation and allergy.In the incidence of allergic contact dermatitis (ACD), the balance between Thl cells and Th2 cells play an important role in regulating, so we verified the effects of ginkgolic acid on related gene expression which can regulate the balance between Thl cells and Th2 cells. These genes included the main cytokines IFN-y that Thl cells secrete and its’transcription factor T-bet, the main cytokines IL-4 that Th2 cells secrete and its’transcription factor GATA-3, in addition, we also studied the impact of ginkgolic acid on ICAM-1 gene expression.The results showed that the expression levels of IFN-γ gene and the transcription factor T-bet gene of treatment group were lower than the model group, but the treatment group of IL-4 gene and transcription factor GATA-3 gene expression amount were higher than the model group. It showed that ginkgolic acid can affect the expression of Thl cells and Th2 cells associated cytokines and transcription factors to regulate the balance of Thl cells and Th2 cells in between, so as to achieve the purpose of treat allergic contact dermatitis (ACD). In addition, the drug group of ICAM-1 gene expression quantity was lower than the model group, which suggested that ginkgolic acid adjust the ICAM-1 gene expression at the same time.In conclusion, the method of water washing combine with silica gel chromatography and alkali extraction method have a good effect of ginkgolic acid purification Ginkgolic acid also have the activity of anti-inflammatory, and its mechanism of action to allergic contact dermatitis(ACD) is related to the balance between Thl cells and Th2 cells and the ICAM-1 gene expression.