| Objective:To investigate the therapeutic effect of femoral artery injection of Ginkgo biloba extract on patients with thromboangiitis obliterans in phase I and II,and to explore the signal pathway of Ginkgo biloba in the treatment of thromboangiitis obliterans through network pharmacology,under the guidance of network pharmacology,the effect and mechanism of Ginkgo biloba on thromboangiitis obliterans were explored through animal experiments,all the above provide ideas for the clinical treatment of thromboangiitis obliterans.Methods:1.Clinical study:Seventeen patients with thromboangiitis obliterans treated with Ginkgo biloba extract(Ginaton injection)were hospitalized in our hospital from November2019 to November 2020.The changes of intermittent claudication distance,skin temperature,ABI and pain were observed one day before treatment and one day after a course of treatment.2.Network pharmacology research:In order to explore the signal pathway of effective components in Ginkgo biloba leaves to play a therapeutic role,the target points of effective components in Ginkgo biloba leaves and related targets of thromboangiitis obliterans were collected and the signal pathway was enriched.3.Animal experiment:Sixty SD rats were randomly divided into four groups: blank group,sham operation group,model group and Ginkgo biloba treatment group.Rats in the blank group,sham operation group and model group were fed with normal saline every day,while rats in Ginkgo biloba treatment group were fed with concentrated solution of Ginkgo biloba every day for 7 days.We observed the changes of rat’s physical signs from the beginning of modeling.After collecting samples,we detected the expression and content of TNF-α,NF-κB,VCAM-1,IκBα protein by immunohistochemistry and Western Blot.Results:1.After a course of treatment with Ginkgo biloba extract injected into femoral artery,the claudication distance of patients increased after treatment,and the difference was statistically significant(P<0.01).After treatment,the skin temperature and ABI of patients were higher than those before treatment,and the difference was statistically significant(P<0.01).The VAS score of patients after treatment was lower than that before treatment,and the difference was statistically significant(P<0.01).2.The network pharmacological research shows that the effective components of Ginkgo biloba leaves may regulate TNF/NF-κB signaling pathway to play a therapeutic role.3.Animal experiment results(1)Changes of physical signs in rats after modeling In this study,the rat model of thromboangiitis obliterans was successfully established.On the first day after the model was established,the rats’ feet became ischemic and dark,and on the third day,the toes of the rats became black and intermittent claudication.On the seventh day,the toes of the affected limbs became dry and black and necrotic.On the third day,the condition of the rats in Ginkgo biloba treatment group was milder than that in the model group,and there was no missing toes in the model group and Ginkgo biloba treatment group.(2)Measurement of TNF-α,NF-κB,VCAM-1 and IκBα expression Immunohistochemical staining was used to observe the expression of TNF-α,NF-κB,VCAM-1 and IκBα in rat femoral artery.The results showed that the positive expressions of TNF-α,NF-κB,VCAM-1 and IκBα in model control group and Ginkgo biloba treatment group were higher than those in blank control group and sham operation group,and the difference was statistically significant(P<0.05);The expressions of TNF-α,NF-κB,,VCAM-1 and IκBα in Ginkgo biloba treatment group were significantly lower than those in model control group(P<0.05).Western Blot showed that the expression levels of TNF-α,NF-κB,VCAM-1 and IκBαin the model control group were higher than those in the blank control group and sham operation group(P<0.05),and the difference was statistically significant.Compared with the blank control group and sham operation group,the expression levels of TNF-α,NF-κB,VCAM-1 and IκBα in Ginkgo biloba treatment group were higher.The difference was statistically significant(P<0.05).Compared with the model control group,the expressions of TNF-α,NF-κB,VCAM-1,IκBα in Ginkgo biloba treatment group decreased after 7days of oral administration,and the difference was statistically significant(P<0.05).Conclusion:Injection of Ginkgo biloba extract into femoral artery can improve the limping distance,skin temperature,ABI and pain of patients with thromboangiitis obliterans.Ginkgo biloba can alleviate the inflammatory reaction of rats with thromboangiitis obliterans,and its clinical efficacy may be related to the regulation of TNF/NF-κB signaling pathway. |
