Observation On The Efficacy And Mechanism Of Qingshi Zhiyang Ointment In The Treatment Of Psoriasis Vulgaris (blood-heat Syndrome

Psoriasis vulgaris,known as "bai bi" in traditional Chinese medicine,is a common chronic inflammatory skin disease mediated by the immune system.It is characterized by scaly,itchy,and erythematous skin lesions,which seriously impair the physical and mental health of patients.External therapy is an important treatment method for psoriasis vulgaris,and traditional Chinese medicine has unique advantages in external treatment.However,there is currently limited research on the external treatment of psoriasis vulgaris with traditional Chinese medicine.Seeking effective topical Chinese medicine preparations and further exploring their mechanisms of action have important practical significance.In recent years,high-mobility group box 1 protein(HMGB1),a widely distributed proinflammatory cytokine in cells,has received much attention.After cells are exposed to stress,damage,or undergo necrosis,HMGB1 can be released from the nucleus and secreted into the extracellular space,where it activates multiple inflammation-related signal transduction pathways by binding to pattern recognition receptors.Recent studies have shown that the binding of HMGB1 to its ligand RAGE activates the NF-κB signaling pathway,triggering a cascade of inflammatory responses that play an important role in the pathogenesis of psoriasis.Intervention in the HMGB1/RAGE/NF-κB pathway may become a promising new anti-inflammatory strategy for psoriasis.Qingshi Zhiyang ointment is a topical traditional Chinese medicine developed by Professor Li Yuanwen based on the experience of the renowned Chinese medicine expert Professor Jin Qifeng.The ointment’s main therapeutic principle is "clearing heat and detoxifying",which is in line with the pathogenesis of psoriasis known as "blood heat toxin accumulation" Our department has accumulated years of clinical experience in treating psoriasis with Qingshi Zhiyang ointment.Animal experiments have also shown that it can inhibit the expression of inflammatory factors such as TNF-α and the expression of RAGE and NF-κB p65 proteins in psoriasis-like mouse skin lesions.However,the exact clinical efficacy of Qingshi Zhiyang ointment in treating psoriasis vulgaris(blood heat syndrome)and its anti-inflammatory effect on psoriasis via the HMGB1/RAGE/NF-κB pathway at the animal level still require further exploration.Therefore,two separate research projects have been initiated to investigate the clinical efficacy of Qingshi Zhiyang ointment in treating psoriasis vulgaris(blood heat syndrome)and its impact on the HMGB1/RAGE/NF-κB signaling pathway.The aim is to elucidate the mechanism by which the ointment improves psoriasis lesions and provide clinical and experimental evidence for its wider application in clinical practice.Part 1 Observation of the Therapeutic Efficacy of Qingshi Zhiyang Ointment for psoriasis vulgaris with Blood Heat SyndromeObjective:To evaluate the clinical efficacy of topical Qingshi Zhiyang ointment for the treatment of psoriasis vulgaris with blood heat syndrome.Methods:A prospective,randomized,double-blind,placebo-controlled study was conducted on 72 patients with psoriasis vulgaris with blood heat syndrome who were treated at the Dongfang Hospital of Beijing University of Chinese Medicine.The patients were divided into treatment group(n=36)and control group(n=36).The treatment group received topical application of Qingshi Zhiyang ointment twice daily,while the control group received placebo ointment(matrix)twice daily.Both groups were also treated with an oral Chinese herbal medicine,Liang Xue Jie Du decoction,which was taken once a day for four weeks.The patients were assessed at baseline,2 weeks and 4 weeks of treatment,and the Psoriasis Area and Severity Index(PASI),Visual Analog Scale(VAS)for itch,Dermatology Life Quality Index(DLQI)and adverse events were recorded.Results:A total of 61 valid cases were collected in this study,including 30 cases in the experimental group and 31 cases in the control group.Before treatment,there were no statistically significant differences between the two groups in terms of gender,age,disease duration,BMI,family history of psoriasis,PASI score,lesion area,erythema,infiltration,scaling score,VAS score,and DLQI(P>0.05),indicating comparability.At 2 weeks of treatment,there were no statistically significant differences between the two groups in terms of PASI score,lesion area,erythema,infiltration,scaling score,VAS score,and DLQI(P>0.05).At 4 weeks of treatment,there were statistically significant differences in PASI score,erythema,infiltration,and VAS score between the two groups(P<0.05),with the experimental group showing lower scores than the control group.There were no statistically significant differences in lesion area,scaling score,and DLQI between the two groups(P>0.05).Within-group comparisons showed statistically significant differences in PASI score,lesion area,erythema,infiltration,scaling score,VAS score,and DLQI between pre-treatment and 4 weeks of treatment(P<0.05).At 2 weeks of treatment,the total effective rate was 50.00%in the experimental group and 19.35%in the control group,with statistically significant differences(P<0.05).At 4 weeks of treatment,the total effective rate was 86.67%in the experimental group and 70.97%in the control group,with statistically significant differences(P<0.05).At 2 weeks of treatment,26.67%of patients(8/30)in the experimental group achieved PASI50,compared to 3.23%of patients(1/31)in the control group,with statistically significant differences(P<0.05).At 4 weeks of treatment,56.67%of patients(17/30)in the experimental group achieved PASI50,compared to 25.81%of patients(8/31)in the control group,with statistically significant differences(P<0.05).At 2 weeks of treatment,6.67%of patients(2/30)in the experimental group achieved PASI75,compared to 0.00%of patients(0/31)in the control group,with no statistically significant differences(P>0.05).At 4 weeks of treatment,10.00%of patients(3/30)in the experimental group achieved PASI75,compared to 0.00%of patients(0/31)in the control group,with no statistically significant differences(P>0.05).During the study period,three cases of adverse reactions occurred in the experimental group and one case occurred in the control group,with symptoms relieved after discontinuation of the medication.Conclusion:Qingshi Zhiyang ointment combined with oral traditional Chinese medicine and placebo ointment combined with oral traditional Chinese medicine are both effective in treating psoriasis vulgaris(blood heat syndrome),and the former shows significant superiority in improving erythema,infiltration,and pruritus of the lesions compared to the latter,and is worth promoting in clinical practice.Part 2 Mechanism of Qingshi Zhiyang ointment’s intervention in psoriasis-like mice by affecting the HMGB1/RAGE/NF-κB pathwayObjective:To observe the intervention effect of Qingshi Zhiyang ointment on psoriasis through the HMGB1/RAGE/NF-κB pathway,and to reveal its anti-inflammatory effects and characteristics in improving psoriatic skin lesions.Methods:Forty-eight male SPF BALB/c mice were randomly divided into a blank control group,a model control group,a traditional Chinese medicine treatment group(Qingshi Zhiyang ointment group),a negative control group(matrix ointment group),and a positive control group(dexamethasone acetate cream group).Psoriasis-like mice were induced using 5%imiquimod,and the corresponding drugs were given for 7 days.Changes in the Psoriasis Area and Severity Index(PASI)score were observed,and mouse skin tissue was collected.Hematoxylin-eosin(HE)staining was used to evaluate the histopathological changes in mouse tissues.Immunohistochemistry was used to detect the effect of Qingshi Zhiyang ointment on the expression of CD3+T cells in mouse skin lesions.Immunofluorescence was used to detect the effect of Qingshi Zhiyang ointment on the distribution and expression of HMGB1.Western blot was used to detect the effect of Qingshi Zhiyang ointment on the expression of pathway-related molecules,including HMGB1,RAGE,NF-κB p65,and p-p65 proteins.Enzyme-linked immunosorbent assay(ELISA)was used to detect the effect of Qingshi Zhiyang ointment on the levels of inflammatory factors such as TNF-α,IL-6,INF-γ,IL-17,and IL-22 in mouse skin lesions.Flow cytometry was used to detect the effect of Qingshi Zhiyang ointment on the proportion of Thl and Th17 cells in mouse skin lesions.Results:HE staining of mouse skin tissue showed that the model group had a significant increase in epidermal thickness,excessive and incomplete keratinization,thickened spinous layer,downward extension of the epidermal papillae,and significant infiltration of inflammatory cells in the dermis.Shallow dermal capillaries were dilated,forming typical pathological changes of psoriasis skin tissue.Compared with the model group,the Qingshi Zhiyang ointment group and dexamethasone group had thinner epidermal layers,less downward extension of the epidermal papillae,and less severe excessive and incomplete keratinization.The infiltration of inflammatory cells in the dermis was also reduced.Measurement of epidermal thickness showed that,compared with the model group,the epidermal thickness of mice in the Qingshi Zhiyang ointment group and dexamethasone group was significantly reduced(P<0.01).Immunohistochemistry experiments showed that the brown precipitate in the dermis of mice in the model group was significantly increased,and a small amount migrated to the epidermis.The brown precipitate in the Qingshi Zhiyang ointment group and dexamethasone group was significantly lower than that in the model group.Compared with the model group,the mean optical density and integral optical density values of CD3 expression in the Qingshi Zhiyang ointment group and dexamethasone group were significantly reduced(P<0.05)by immunohistochemistiy.Flow cytometry analysis showed that,compared with the model group,the proportion of IFNγ+Thl cells and IL17A+Thl7 cells in CD4+T cells was significantly reduced in the Qingshi Zhiyang ointment group and dexamethasone group(P<0.01).ELISA experiments showed that,compared with the model group,the levels of TNF-α,IL-6,IFN-γ,IL-17,and IL-22 were significantly reduced in the Qingshi Zhiyang ointment group and dexamethasone group(P<0.01).Immunofluorescence detection showed that,compared with the model group,the mean optical density and integral optical density values of HMGB1 expression in the Qingshi Zhiyang ointment group and dexamethasone group were significantly reduced(P<0.01).Western blotting showed that,compared with the model group,the relative content of cytoplasmic HMGB1 in the Qingshi Zhiyang ointment group and dexamethasone group was significantly reduced,and the relative content of nuclear HMGB1 was significantly increased.The ratio of cytoplasmic to nuclear HMGB1 relative content was significantly reduced(P<0.01).Compared with the model group,the relative content of RAGE protein and the expression ratio of NF-κB p-p65/p65 protein in the Qingshi Zhiyang ointment group and dexamethasone group were significantly reduced(P<0.01).Conclusion:Qingshi Zhiyang ointment can improve psoriasis-like skin lesions in mice,reduce epidermal thickness and inflammation.Its mechanism may be related to the inhibition of HMGB1 expression and translocation in skin tissue,which regulates the HMGB1-mediated RAGE/NF-κB signaling pathway and inflammatory cascade reaction,inhibits the activation of Thl and Th17 cells,and reduces the expression of inflammatory factors TNF-α,IL-6,IFN-y,IL-17,and IL-22.