| Psoriasis vulgaris, known as "Baibi" in Traditional Chinese Medicine, is a chronic, recurrent and inflammatory disease of the skin with erythema and sliver scaling. It is a complex multifactorialdisease and its pathogenesis and precise mechanism remain poorly understood. Nowdays, psoriasis vulgaris is mainly recognized as a polygenic disorder linked with gene and environment factors. The activation of T cell is one of the most crucial step in the pathogenesis of psoriasis, meanwhile, which is regulated by cytokines and multiple signaling pathways. Some studies have determined that the psoriasis vulgaris has a close association with the Notch signaling pathway. It has also been demonstratedthat the expression of Notch receptors weredetected in the dermal layer. InT-lymphocytes in the peripheral blood, the expression of Notch-1 and Notch-2 receptors increased, and the ability to transductthe activition signal was also enhanced. As the target gene in the downstream of Notch pathway, Hes-1 gene can indirectlyreflect the expression of Notch pathway. Therefore, The way of regulating Notch signaling pathway may be a new therapeutic approach for psoriasis.Based on TCM theory, "Blood heat" is the main cause of psoriasis vulgaris. According to the core pathogenesis,Professor Yan-ping Baihas worked out the Heat-clearing and blood-cooling recipe, which also known as "Quyin granula"for patients with blood-heat type psoriasis vulgaris. Thisrecipe consists of many herbs having the effect of clearing away heat, cooling blood and detoxifying. Its clinical therapeutic effect has been confirmed by some clinical trials and animal experiments.However, the specific mechanism and the target of Heat-clearing and blood-cooling recipe are not clear.Objective:The purpose of this paper is to reveal the expression level of Notch-1 receptor in CD4+T-lymphocytes in peripheral blood of patients withblood-heat typepsoriasis, to explore the effect of Heat-clearing and blood-cooling recipefor the activation Notch signaling pathway in vitro, and to clarify the mechanism of Heat-clearing and blood-cooling recipein psoriasistreatment through regulating Notch signaling pathway.Methods:The CD4+T-lymphocytesof patients withblood-heat typepsoriasiswere employed in this study. RT-PCR and Western blotting were performed to detect the mRNA and protein expressions of Notch-1 receptor inCD4+T-lymphocytes.The time and concentrations of CD4+T-lymphocytes activation after stimulation of Phytohemagglutinin (PHA) were observed. After screening the different concentrationsof Jagged-1 fusion protein, the optimal concentration ofactivation Notch signaling pathway in vitro was established. Serum containing Heat-clearing and blood-cooling recipe(low, moderate,high dose) andnormal saline were prepared. The Hes-1 mRNA expressionin each group containing serum was detected by RT-PCR. The proliferation and inhibition ofCD4+T-lymphocytes were evaluated using CCK-8 test.The TNF-a, IL-2, IFN-y expression of the cell supernatant in each groupwere detected by ELISA.Results:(1)There was significant difference between patients withblood-heat typepsoriasis and healthy people in Notch-1 receptor mRNA and protein in CD4+T-lymphocytes (P<0.01).(2)48h after PHA stimulation on CD4+T-lymphocytes, the proliferation and division of Tlymphocytes in lOug/ml concentration of PHA were better than that in 5ug/ml concentration, there was significant differencebetween lOug/ml and 5ug/ml. There was a statistically significantly difference between the two groups(P<0.01).(3)When Jagged-1 fusion protein in 0.5ug/mlconcentration stimulatingCD4+T-lymphocytes for 48h, the downstream of Notch-1 signaling pathway can be activated.Meanwhile, both the Hes-1 mRNA expression and the proliferation of CD4+T-lymphocyte were significantly higher than that in control group(P<0.05). These results suggest activation Notch signaling pathway was establishedin vitro.(4) The Hes-1 mRNA expression in the moderate andhigh doseHeat-clearing and blood-cooling recipegroups were markedly lowered than that in model group (P<0.05). Two groupscould reduce the secretion ofTNF-a, IFN-y secretion in CD4+T-lymphocytes, and increase the secretion of IL-2.There were significantly different compared with low dose group(P<0.05).Conclusion:Notch-1 receptor in Notch signaling pathway were in a high expression status in CD4+T-lymphocytes of patients with blood-heat type psoriasis. The Jagged-1 fusion protein in 0.5ug/ml concentration could activate Notch-1 signaling pathway, and promote proliferation and division of CD4+T cell in the peripheral blood in patients with blood-heat type psoriasis. These results suggested that Notch signaling pathway may play a role in regulating the proliferation and division of T cell in psoriasis. The Heat-clearing and blood-cooling recipe could inhibit the activation of Notch-1 signaling pathway, thereby reduce the secretion of TNF-a, IFN-y in CD4+T-lymphocytes, and increase the secretion of IL-2. Meanwhile, the effect in moderate and high dose Heat-clearing and blood-cooling recipe last longer than that in the low dose group. |
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