Clinical Study On The Effect Of Heat-clearing And Dampness-removing Therapy On Psoriasis And Its Effect On The Expression Of JAK/STAT3 Pathway In Keratinocytes

Objective:The clinical evidence of heat-clearing and dampness-removing therapy in the treatment of psoriasis vulgaris is sufficient,but the reported results are not consistent.Therefore,this study is based on the guidance of evidence-based medicine,through the Meta analysis of the published randomized controlled trials of heat-clearing and dampness-removing therapy in the treatment of psoriasis vulgaris,combined with the clinical observation of Qingre Lishi Yin,an effective prescription in our department,in the treatment of psoriasis.It is expected to analyze the efficacy of heat-clearing and dampness-removing therapy in the treatment of psoriasis vulgaris according to the existing research data,and evaluate the efficacy and advantages of this method subjectively and objectively.Then,through the culture of Ha Ca T cells and the establishment of psoriatic mouse model,we could analyze the effect of Qingre Lishi Yin on Ha Ca T cells and tyrosine kinase protein JAK/STAT3 pathway in psoriatic lesions,and explain its regulatory mechanism,which is helpful to further explore the mechanism of heat-clearing and dampness-removing therapy in the treatment of psoriasis and enrich the theory of TCM syndrome differentiation in the treatment of psoriasis.Methods:1.Meta analysis:adopt the retrieval method of the combination of subject words and free words,formulate the search words from three aspects:target diseases,intervention measures and research types,and adjust the retrieval strategy according to the specific database.The randomized and controlled trials in English database(Pub Med,EMBASE,Web of Science))and Chinese database(CNKI,VIP,Wan Fang)about Qingre Lishi Yin or combined with other therapies in the treatment of psoriasis were searched.After reading,screening and extracting effective information,risk assessment tools were used to evaluate the quality,and Rev Man5.3.0 was used to analyze the data of randomized controlled trials that met the inclusion criteria and exclusion criteria.2.Clinical study:using randomized controlled trial(RCT)design,102 patients with psoriasis vulgaris were randomly divided into traditional Chinese medicine group(n=52)and proprietary Chinese medicine group(n=50).The traditional Chinese medicine group was given Qingre Lishi Yin orally,the proprietary Chinese medicine group was given Xiaoyin granule orally,and both groups were treated with carpotriol ointment as the basic treatment drug for a total of 8 weeks.The area and severity index of psoriasis(PASI)was taken as the main observation indexes,while the skin lesions occupying body surface area(BSA),itching visual analogue score(VAS),dermatosis quality of life index(DLQI)and safety were taken as the secondary observation indexes.The effective rates of the two groups were calculated respectively.3.The vitro cell experiment:On the basis of previous studies,this study further studied and analyzed the effect of Qingre Lishi Yin on JAK-STAT3 pathway in Ha Ca T cells.Ha Ca T cells were cultured in complete medium containing different concentrations of Qingre Lishi Yin.After 48 hours of intervention,the protein expressions of JAK1,JAK2,P-ERK and P-STAT3 were detected by Westernblot.The m RNA expression of inflammatory cytokines IL-6,CXCL1,TNF-αand STAT3 was measured by real-time PCR.4.The vivo animal experiment:50 BALB/c male mice were randomly divided into blank control group(Control),imiquimod cream(IMQ)induction+normal saline intragastric administration group,IMQ induction+Qingre Lishi Yin high dose gastric perfusion group(QRLSY-H),IMQ induction+Qingre Lishi Yin medium dose gastric perfusion group(QRLSY-M),IMQ induction+Qingre Lishi Yin low dose gastric perfusion group(QRLSY-L),with 10 mice in each group.The skin lesions of psoriatic mice were evaluated by PASI score and HE staining.The expressions of proliferating cell nuclear antigen(Ki67),PCNA and P-STAT3 in signal pathway were detected by immunohistochemistry.The expressions of JAK1,JAK2,TYK2,STAT3,STAT5 and P-STAT3 were detected by Westernblot,and the expressions of STAT3,IL-6,IL-22,IL-17A and IFN-γm RNA in tissues were quantitatively detected by real-time fluorescence.Results:1.Meta analysis:a total of 1189 cases and 16 literatures(including 6 journals and 10 dissertations)were included in this study,all of which were randomized controlled clinical trials,and the quality of the literatures was acceptable,so this Meta analysis has high credibility.The experimental group and the control group were analyzed by Meta from the aspects of clinical effective rate,clinical cure rate,PASI score,adverse reactions and recurrence rate.The results of PASI score Meta analysis showed that the study had high heterogeneity(P<0.00001,I~2=98%).The random effect model was used to carry out the combined statistics,Z=2.02,P<0.05,which was statistically significant.The heterogeneity of Meta analysis of clinical effective rate,clinical cure rate,adverse reaction and recurrence rate was low(I~2<50%).Using fixed effect model,the difference was statistically significant(P<0.05).2.Clinical study:In the traditional Chinese medicine group,the cure rate was11.54%,the markedly effective rate was 48.08%,the effective rate was 26.92%,the ineffective rate was 13.46%,and the total effective rate was 86.54%.In the proprietary Chinese medicine group,the cure rate was 2.00%,the markedly effective rate was 34.00%,the effective rate was 42.00%,the ineffective rate was 22.00%,and the total effective rate was 78%.The total effective rate,cure rate and markedly effective rate of Qingre Lishi decoction in the treatment of psoriasis were higher than those in Xiaoyin granule group,while the effective rate and inefficiency in Xiaoyin granule group were lower than those in Xiaoyin granule group.After treatment,the scores of PASI,BSA,DLQI and VAS were significantly decreased in both the traditional Chinese medicine group and the proprietary Chinese medicine group,and the scores in the traditional Chinese medicine group were more significantly lower than those in the proprietary Chinese medicine group.Three patients in the traditional Chinese medicine group had severe abdominal pain and diarrhea after taking medicine,which disappeared after drug withdrawal,while there were no adverse reactions in the proprietary Chinese medicine group.3.Vitro cell experiment:compared with the group without Qingre Lishi Yin,Qingre Lishi Yin could reduce the expression of JAK1,JAK2 and P-STAT3 in Ha Ca T cells,and the inhibitory effect was concentration-dependent,but there was no significant difference in the expression of phosphorylated extracellular regulated protein kinase P-Erk.Qingre Lishi Yin could also inhibit the expression of IL-6,TNF-αand STAT3m RNA in Ha Ca T cells,and the inhibitory effect was dose-dependent,especially in high concentration Qingre Lishi Yin group.However,Qingre Lishi Yin had no significant effect on the expression of CXCL1 m RNA,and the difference was not statistically significant(P>0.05).4.Animal experiment in vivo:Qingre Lishi Yin could significantly improve the symptoms of erythema,scale and infiltration on the back of psoriatic mice induced by imiquimod,and reduce the PASI score,compared with IMQ group,Qingre Lishi Yin could reduce the thickness of keratinocytes and histopathology of epidermis in psoriasis-like mice.The results of immunohistochemical experiment showed that Qingre Lishi Yin could significantly reduce the number of PCNA and Ki67 positive cells in the epidermis of psoriatic mice.Qingre Lishi Yin could also reduce the expression of STAT3,P-STAT3,JAK1 and JAK2 protein in keratinocytes of psoriatic mice.The difference was statistically significant,and the concentration of Qingre Lishi Yin was dependent,but had no effect on the expression of STAT5 and TYK2.Qingre Lishi Yin could also inhibit the expression of IL-6,IL-22,IL-17A,IFN-γand STAT3m RNA in skin tissue of mice,and the difference was statistically significant.Conclusion:1.The results of Meta analysis show that the method of clearing heat and removing dampness can improve the effective rate of psoriasis,improve the clinical symptoms of patients with psoriasis and reduce the occurrence of adverse reactions,but limited by the inclusion of the literature,this conclusion still needs to be further verified.2.Clinical observation shows that Qingre Lishi Yin can significantly improve the clinical symptoms of patients with psoriasis vulgaris,improve the quality of life of patients,improve the symptoms of itching,and high safety.3.Cytological experiments show that Qingre Lishi Yin may play a role in the treatment of psoriasis by inhibiting the expression of JAK/STAT3 signal pathway protein in Ha Ca T cells and the secretion of IL-6 and TNFαinflammatory cytokines.4.Animal experiments show that Qingre Lishi Yin can reduce the expression of inflammatory cytokines and improve psoriasis-like inflammation induced by IMQ,which may play a role in the treatment of psoriasis by inhibiting the expression of JAK/STAT3signal pathway.