| Background:Traditional Chinese medicine has a history of treating psoriasis for over a thousand year,and "treating from the aspect of blood" is one of the most popular dialectical thoughts for modern Chinese physicians in treating psoriasis.Psoriasis vulgaris,according to the thoughts of "treating from the aspect of blood",is basically divided into three types,which respectively belongs to three different syndromes:blood-heat,blood-dryness and blood stasis.Yangxue Jiedu Decoction,whose clinical effect is definitely positive,is a classical Chinese prescription in the treatment of psoriasis vulgaris with blood dryness syndrome.Patients with psoriasis vulgaris are usually annoyed by red maculo-papulaes,squamae,and probably with pruritus at the same time.The pathogenesis of this disease,which is rather complex,is related to many aspects such as autoimmune,inflammation and cell autophagy,and apoptosis.The main pathological changes of psoriasis are keratinocyte proliferation,inflammatory cell infiltration and capillary angiogenesis.Apoptosis is a way of active programmed cell death,which plays a role in psoriasis vulgaris through anti-proliferation.In the skin lesions of patients with psoriasis,the apoptosis was found significantly abnormal.The levels of anti-apoptotic proteins such as VEGF,PI3K,Akt and Survivin were significantly up-regulated,and the apoptoticproteins such as Fas,FasL and bax showed up and reached a rather high level.Whereas in the lesions of normal people,these apoptotic proteins were not obviously detected.These changes in anti-apoptotic proteins and apoptotic proteins strongly confirm that apoptosis is involved in the pathogenesis of psoriasis and inducing apoptosis to inhibit proliferation is an effective method for the treatment of psoriasis.Previously,members of our research group found that Yangxue Jiedu Decoction could improve psoriasis-like lesion symptoms in mice by regulating the VEGF/PI3K/Akt/Survivin pathway in the mouse model of psoriasis vulgaris.Therefore,it is presumed that Yangxue Jiedu Decoction can inhibit cell proliferation and improve psoriasis lesion symptoms by regulating cell apoptosis and related protein expression.Objective:Preliminary study on the changes caused by Yangxue Jiedu Decoction on PASI score,histopathology,apoptosis,and Cyclin D1 in skin lesions of psoriasis patients with blood dryness syndrome and its intervention on the changes of Survivin mRNA in serum;comparison of blank group,model group,chinese medicine group and positive control group;preliminary study on Yangxue Jiedu Decoction's function in imiquimod model of psoriasis-like mice on skin lesion morphology,histopathology,apoptosis,those related proteins(bcl-2,bax,Caspase-3,Cyclin D1)and VEGF,Survivin mRNA in the serum.Observations are mainly focused on the changes of apoptosis and the related proteins in psoriasis vulgaris participants and mouse models before and after the treatment with Yangxue Jiedu Decoction.And it will be explored in this paper whether Yangxue Jiedu Decoction can interfere with apoptosis and the possible mechanism of this decoction in treating psoriasis.Methods:1 Clinical trial:Thirty-three subjects with psoriasis vulgaris of blood dryness syndrome who fit the inclusion and exclusion criteria were enrolled.They were chosen from the patients treated at the Department of Dermatology,Beijing Hospital of Traditional Chinese Medicine,affiliated to Capital Medical University,from 2018 to 2020.And inclusion of 11 healthy subjects from social recruitment.After entering the group,the subjects with psoriasis vulgaris of blood dryness syndrome were treated with Yangxue Jiedu Decoction granules for 8 weeks.If the skin of the subjects was seriously dry,they could be applied with white vaseline or silicon cream to moisturize the skin.Blood and skin specimens were collected after their permission on the day of getting in and out of the group.After communication,we collected 20 cases of skin lesions before treatment,7 cases of skin lesions after treatment and 7 cases of normal skin.Healthy subjects only kept blood samples.Separated from theblood samples after centrifugation,the supernatants were stored in a negative 80? freezer.And then detect the expressions of Survivin mRNA in it by Quantitative Real-time PCR(qPCR).Paraffin sections are made from the skin tissues after being soaked in 10%formaldehyde solution.And then the apoptosis and the positive expression of Cyclin D1 were determined by TUNEL and immunohistochemical methods.2 Animal experiment:Forty BALB/c mice were randomly divided into four groups with 10 mice in each group.The four groups includes a normal control group,a model group,a Yangxue Jiedu Decoction group(namely the traditional Chinese medicine group),and a cyclosporine control group(also named the positive control group).After a five-day adaptive feeding,all the mice were hair-shaved.Except for the normal control group,mice of the other groups were used to establish models of psoriasis-like lesions,which lasted for six days and formal experiments were to be carried out after the success in modeling.On the 7th day of modeling,the eyeball blood of mice was taken to collect blood samples and the mice were executed by cervical vertebra dislocation method.Then cut the back skins of the mice and fix them with 4%paraformaldehyde for the preparation of paraffin sections.The expressions of VEGF and Survivin in serum were detected by Real-time PCR.And the positive expressions of bcl-2,Bax,Caspase-3 and Cyclin D1 in the skin lesions of mice were determined by TUNEL and immunohistochemical methodsResult:1 Clinical trial:? The PASI score was decreased at the fourth week after the treatment with Yangxue Jiedu Decoction(P<0.05).? In this experiment,the total effective rate of Yangxue Jiedu Decoction in the treatment of psoriasis vulgaris with blood dryness syndrome was 72.73%;Compare the normal skin of subjects with blood dryness syndrome in psoriasis vulgaris,the epidermis thickness of psoriasis vulgaris patients with blood dryness syndrome was significantly thickened by HE staining(P<0.05).After 8 weeks of Yangxue Jiedu Decoction treatment,the epidermis thickness was significantly thinner than that before treatment(P<0.05).? Compared with the normal skin,the apoptosis in the lesion of psoriasis blood dryness syndrome was inhibited(P<0.05),and the apoptosis level in the lesion was increased after treatment(P<0.05).?The positive expression of Cyclin D1 in the lesions of psoriasis vulgaris with blood dryness syndrome increased,and the positive rate decreased slightly after treatment,but there was no statistical difference(P>0.05).?The expression of Survivin mRNA in serum of psoriasis vulgaris patients with blood dryness syndrome was higher than that of healthy control group,but there was no statistical significance(P>0.05).There was no statistical significance in the expression of Survivin mRNA in serum of patients before and after treatment(P>0.05).2 Animal experiment:?The epidermises of mice in the model group,the positive control group and the traditional Chinese medicine group were significantly thicker than those in the blank group(P<0.05),and the epidermises of mice in the positive control group and the traditional Chinese medicine group were significantly thinner than those in the model group(P<0.05).? The apoptosis rate of the skin lesions of mice in the blank group was the highest,with statistical differences(P<0.05).While there were no significant differences among the model group,the positive control group,and the Chinese medicine group(P>0.05).?There were no significant differences in Cyclin D1 expression among the blank group,the model group,the positive control group and the Chinese medicine group(P>0.05)However,from the perspective of the positive rate,the positive expression of Cyclin D1 in the model group was higher than that in the blank group,while in the Chinese medicine group it was lower than that in the other three groups.?There was no significant differences in the positive expression of bcl-2 and bax in the skin lesions of the blank group,the model group,the positive control group and the traditional Chinese medicine group(P>0.05),However,from the perspective of positive rate,the positive expression rates of bcl-2 and bax in the model group,the positive control group and the traditional Chinese medicine group were all higher than those in the blank group,and the positive expression rate in the traditional Chinese medicine group was lower than that in the model group.?The expressions of Caspase-3 protein in skin lesions of blank group,the model group,the positive control group and the Chinese medicine group were negative,but from the average score,the expression of Caspase-3 protein in model group was the lowest,and the scores of the Chinese medicine group and the positive control group were higher than those of the blank group and the model group.?The expression of serum VEGF mRNA in the model group and the positive control group were higher than that in the blank group(P>0.05).The expression of serum VEGF mRNA in Chinese medicine group was lower than those in the model group and the positive control group,but there were no significant differences(P>0.05).?Compared with the blank group,the expressions of Survivin mRNA in the serum of mice in the model group and the positive control group were significantly increased(P<0.05).There were no significant statistical differences between the model group and the positive control group(P>0.05).Compared with the model group and the positive control group,the expression of Survivin mRNA in serum of the Chinese medicine group was decreased(P<0.05),but there was no significant difference campared with the blank group(P>0.05).Conclusion:Yangxue Jiedu Decoction may inhibit cell proliferation by down-regulating VEGF mRNA expression and inhibiting Survivin mRNA expression.In addition,Yangxue Jiedu Decoction may promote the apoptosis in the skin lesions of psoriasis vulgaris patients with blood dryness syndrome,but the intervention effect of Yangxue Jiedu Decoction on the expression of bcl-2,bax and Caspas-3 proteins is still unclear.It is necessary to further expand the sample to clarify the relationship. |
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