| Background:Psoriasis, also known as "Niu Pi Xuan", or called "Bai Bi" in Traditional Chinese Medicine (TCM), currently has been regarded as a kind of chronic skin disease of inflammation, relatively appeared in clinic with erythema scale as its main clinic symptom. It often occurs on skin and joints and easily relapses. The incidence of psoriasis is gradually increasing in recent years; however, the specific causes of psoriasis are still unclear. Most experts believe psoriasis is a genetic disease related to multiple genes, which is caused and affected by the interaction of multiple factors, such as genetic and environmental factors. During the whole progress of disease, it is influenced and regulated by multiple complex cytokine networks and signaling pathways.Notch signaling pathway consists of Notch acceptors, Notch ligands and CSL DNA-binding protein. It is widely distributed in the surface of multiple cells, such as hematopoietic stem cells, embryonic stem cells, lymphocytes, and vascular endothelial cells. Some studies show that Notch signal is activated in the dermis, and it can promote the onset of psoriasis by influencing the development and activation of T cells and regulating the secretion of cytokine. Some studies indicate that there is high expression of Notch and its ligand in the surface of keratinocyte and other epidermal cells and the specific activation of Notch signaling plays an important role for maintaining the skin cells’ renewal and differentiation. Therefore, abnormal Notch signals possibly help to reveal the pathogenesis of psoriasis and other skin diseases.To date, there is no domestic or overseas research exploring certain medications for the treatment of psoriasis by regulating the Notch signaling pathway, and research on the mechanism of the treatment is even rarer. Therefore, it is extremely necessary to conduct an intervention study from immune regulation perspective to treat psoriasis by regulating Notch signaling pathway. Blood-heat psoriasis, equivalent to the initial stage or acute stage of psoriasis, is the most typical and common type of psoriasis in clinical practice. Qu Yin Granules is a self-prescribed prescription by Doctoral Supervisor and Professor Bai Yanping. Its essential components consist of Chinese herbal medicines with functions of clearing heat, cooling blood and resolving toxins. We has confirmed its beneficial effects on psoriasis by previous clinical studies and animal experiments. However, the specific mechanism and the action target are still unclear. This study aims at probing whether the Notch signal transduction molecules play a role or not in blood-heat psoriasis from the perspective of molecular biology. Understanding of the active situation of the transduction molecules will support future research on the regulation of Notch signaling pathway by clearing-heat and cooling-blood medications, which will lay a foundation for revealing the whole mechanism of the treatment.Purposes:The purposes of this research are to reveal the expression of Notch signaling molecules (Notch1-4, Deltl, Delt3-4and Jagged1-2) in mononuclear cells of blood-heat psoriasis patients, and to objectively evaluate the relationship between the psoriasis severity and the expression of Notch signaling molecules, and to explore target protein molecules for the regulation of Notch signaling pathway.Methods:27blood-heat psoriasis patients confirmed under the guidance of TCM theory were allocated in the experimental group and graded with PASI scores, with27health participants as control. We collected the peripheral blood samples of all participants to extract their mononuclear cells. Real-time fluorescent quantitation PCR method was used to detect the expression levels of mRNA of Notch signaling molecules of all participants. Flow cytometry was used to record expressions of the mean fluorescence intensity (MFI) of Notch signaling molecules in6blood-heat psoriasis patients and5healthy participants. Western Blotting was used to detect the expression of proteins related to Notch signaling molecules of30participants in experimental group and30healthy participants in control group.Results:(1)Compared with healthy participants, blood-heat psoriasis patients had a marked increase of mRNA level of Notch2in mononuclear cells. There was a significant difference between groups, P=0.036(P＜0.05). The expression was positively correlated with PASI scores.(2)Compared with healthy participants, blood-heat psoriasis patients had an obvious decrease of mRNA level of Notch3in mononuclear cells. There was a significant difference between groups, P=0.008(P＜0.01). The positive correlation between the expression and PASI scores was not detected.(3)Compared with healthy participants, there were no specific changes of Notchl, Notch4, Jaggedl-2, Deltal and Delta3-4in blood-heat psoriasis patients. There was no significant difference between them.(P＞0.05)(4)There was significant statistical difference (P＜0.01) for Notch2MFI between experimental group and healthy group. The PASI correlation analysis showed that the Notch2’s flow MFI expression was independent of the psoriasis severity.(5) There was no statistical significance between the MFI of Notchl and Notch3of experimental group and Notchl MFI of healthy group.(6) Western Blotting showed that there was an obvious increase of Delta4protein expression, with significant statistical difference (P＜0.01). The PASI correlation analysis showed that Delta4protein was independent of the psoriasis severity.(7)Western Blotting showed that there was no difference (P＞0.05) for Notchl or Delta4protein expression between experimental group and healthy group.Conclusions:The mRNA and protein expressions of Notch2of blood-heat psoriasis patients were higher than healthy participants, and the higher of mRNA level of Notch2, the severer of psoriasis, which indicates the direct relation between psoriasis severity and the activity level of Notch2. As Notch signaling molecule ligand, Delta4has directly participated in the course of blood-heat psoriasis. Though psoriasis participants had a decrease of Notch3mRNA expression, compared to healthy people, it cannot be used as diagnostic standards for psoriasis. Notch signaling pathway directly has participated in the onset and development of blood-heat psoriasis. |
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