Research On The Preparation Process Of Ginkgo Acid And Its Antibacterial Activity

In this paper, we studied the the separation, purification and antibacterial activity of Ginkgo acid. Through a series of purification explore and experiment, we found that the use of membrane separation and purification technology to purify ginkgo acid was a good way. Use of Staphylococcus aureus and Propionibacterium acnes in vivo and in vitro inhibitory effect on acid to clarified the inhibitory effect of Ginkgo acid. The following is the main content and result:1、The separation and purification of Ginkgo acid. Ethanol extraction method to extract ginkgo acid, then membrane separation was experimented. By orthogonal (the main factor is the time, the membrane molecular weight, the pressure) optimum condition of membrane separation and purification. Optimal conditions for the use of ginkgo acid purification,the purity of ginkgo acid increased from 33.9% to 77.82%.2、In vitro antibacterial activity of Ginkgo acid. In this paper, we studied the antibacterial effect of ginkgo acid by both in vitro and in vivo. Measured by filter paper method Ginkgo acid on two types of bacteria (Staphylococcus aureus and Propionibacterium acnes) has a strong inhibitory effect, use the dilution method to Measured the minimum inhibitory concentration (MIC), the MIC were 137.5 μg/ml and 550.0.0μg/ml, After extended period of bacterial growth and the logarithm of Joining ginkgo acid, bacterial growth is inhibited. Through a series of microscopic observation (ordinary optical electron microscopy, scanning electron microscopy SEM, transmission electron microscopy TEM) studies have shown that ginkgo acid primarily by disrupting the bacterial cell membrane to achieve the inhibitory effect. Polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the bacterial proteins, the results showed that the amount of reduction in bacterial proteins,Found that ginkgo acid destroyed the cell membrane of bacteria makes the cell contents leaked, protein outflow.In vivo antibacterial activity of Ginkgo acid. By subcutaneous injection of Staphylococcus aureus and Propionibacterium acnes in Kunming mice and golden hamsters in vivo respectively, establish an animal model of skin infection. Set four groups, Model group, positive control group, low-dose ginkgo acid group, ginkgo acid high-dose group, positive control group (Bactroban, viaminate vitamin E cream), continuous smear 7 days, The results showed that two model mice with smear ginkgo acid, the lesion area were significantly reduced, the treatment group’ skin scores were better than the model group, lesions were significantly improved conditions, all these description that Ginkgo acid can better treat bacterial infections of the skin acne, skin ulcers and inflamed repaired.3、Histological observation showed that the nuclei of skin cells infected with bacteria messy arrangement, inflammatory cell infiltration, telangiectasia; ginkgo acid treatment group aligned nuclei tend structured, reduction of inflammatory cell infiltration, capillary become normal. Description that ginkgo acid have a good therapeutic effect on the Staphylococcus aureus and Propionibacterium acnes infection.4、Molecular mechanism of action of the Antibacterial Activity. Detected the affect of ginkgo acid treatment on the skin infection model related gene expression. When the skin was infected by the bacterial, machine experience spontaneous generation of a series of natural immune system to defend the body produces inflammation. Genes involved have Toll-like receptor 2 (TLR-2), Toll-like receptor 4 (TLR-4), interleukin-1β (IL-1β), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α). The results showed that Toll-like receptor 2 (TLR-2), Toll-like receptor 4 (TLR-4), interleukin-1β (IL-1β), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α) gene expression in the administered group was significantly lower than the model group, but higher than the positive control group. Description ginkgo acid has a role in reduced bacterial infection inflammatory cytokines, induced innate immune system may be a good treatment of the two bacterial infections of the skin inflammation.In summary, the use of membrane separation technology can be a higher concentration of ginkgo acid and little loss. Ginkgo acid has a good inhibition effect on Staphylococcus aureus and Propionibacterium acnes, and to treat skin inflammation due to the two bacterial infection. The mechanism related to the reduction of skin inflammatory cytokine expression which caused by the bacterial infection.