Clinical Observation On The Treatment Of Vitiligo With Baibo NO.1 Recipe And Mechanism Research On Complannatuside And Catalpol For Melanocytes Ferroptosis

Objectives1.To observe the clinical efficacy of Bai Biao No.1 Recipe on the treatment of vitiligo and its mechanism of action in the treatment of vitiligo.2.To apply the TCM heritage aid platform to analyze the pattern of vitiligo medication.3.Applying network pharmacology and molecular docking to analyse the commonly used drugs for vitiligo and discover the factors associated with iron death and vitiligo development.4.Construct ferroptosis states in melanocytes using RSL-3,Observing melanocyte changes from the perspective of ferroptosis,refining the pathogenesis of vitiligo and exploring the targets of action of Complannatuside and Catalpol.Methods1.Clinical trial: 60 cases of clinically diagnosed vitiligo patients were collected and randomly divided into two groups.The teratmental group was given Bai Biao No.1 Recipe orally,and the control group was given Bai Ling tablets orally,and the conventional treatment was compound kalizarin tincture rubbed externally.The clinical efficacy was determined at the end of the study.2.Analysis of vitiligo medication pattern: Patients with vitiligo previously treated at the Affiliated Hospital of Shandong University of Traditional Chinese Medicine were included in this study,the enclosed patients were collected and also the patients’ prescriptions were processed according to the format requirements of the TCM Heritage Assist Platform V 3.0.3.Network pharmacology and molecular docking part: network analysis of vitiligo and Complannatuside and Catalpol by applying relevant databases and software,and molecular docking of small molecules and target proteins.4.Cellular experiments: Normal human melanocytes were subjected to different concentrations of iron death inducers RSL-3,Complannatuside and Catalpol.The experiments were divided into as control group,RSL-3 group,RSL-3+Complannatuside and group,Catalpol +RSL-3 group and Complannatuside and Catalpol +RSL-3 group.The melanocytes were also observed and tested accordingly.Results1.Clinical findings: Bai Biao No.1 Recipe had significant advantages over the control group in terms of re-colourisation rate and efficacy.Bai Biao No.1 Recipe was more effective on the face and neck and trunk compared to the lesions on the extremities.2.Analysis of vitiligo medication pattern: semen astragali complanati and radix rehmanniae were the commonly used combination.3.Network pharmacological study results: TP53 is present in both iron death signaling pathway and vitiligo pathogenesis,molecular docking results show that all small molecules can enter the active center of the target protein.4.Results of cellular experimental studies:(1)Complannatuside and Catalpol antagonize RSL-3-induced melanocyte growth inhibition.(2)The mitochondrial morphology and dysfunction of melanocytes under oxidative stress were improved by Complannatuside and Catalpol.(3)Complannatuside and Catalpol reduced RSL-3-induced iron ion levels and melanocyte ROS(intracellular,lipid).(4)Complannatuside and Catalpol protected melanocytes by increasing SL C7A11 and GPx4 and decreasing TP53 and TFR1 expression.Conclusion1.The efficacy of Bai Biao No.1 Recipe on the treatment of stable vitiligo is definite with no adverse effects.2.Semen astragali complanati and radix rehmanniae are the main combinations for the treatment of vitiligo.3.The TP53 is the key protein in vitiligo,molecular docking results show that all small molecules can enter the active center of the target protein.4.Complannatuside and Catalpol were able to improve melanocytes in a state of oxidative stress with the involvement of iron death induced by RSL-3.