| 1. BackgroundPsoriasis is a common and frequently-occurring dermatology, traditional Chinese medicine is one of the advantages method for the treatment of disease. In recent years, with the increase of air pollution and social pressure, the trend and incidence rates of this disease have increased every year. "Qingrehuoxuejiedufufang (QHJD)", which is a one of the clinically effective therapy formulae for this disease over past 30 years. It belongs to clinical subjects of "blood heat syndrome" to clear away for the treatment of psoriasis, this formulae can effectively promote regression of lesions, prolong remission and reduce recurrence. Due to the complexity of Chinese formulae medicine in the multi-components, hence the mechanism of QHJD formulae for the treatment of psoriasis is not clear, this experiment is with aim to study this area for the illustration its mechanism in cell or molecular.2. ObjectiveTo observe the decoction of Qing-Re-Liang-Xie-Jie-Du (QHJD) on human immortalized keratinocyte cell (HaCat) proliferation, apoptosis, secretion of inflammatory cytokines and influence on the related apoptotic genes. Based upon those to investigate the mechanism of QHJD for the treatment of psoriasis with regarding to ideas and guidance for the clinical and research.3. Material and Method3.1 The means of CCK-8, flow cytometry, RT-PCR and Western Blot were used to observe QHJD with effect on Hacat cell proliferation, apoptosis, Bcl-xl and Bax expression. The enzyme-linked immunosorbent assay (ELISA), RT-PCR and Western Blot was applied to observe the QHJD influence on the supernatant of Hacat cell factors such as cytokines IFN-γ, IL-1β, IL-2, IL-6,1L-8 secretion and expression of p38.3.2 The means of CCK-8, flow cytometry, RT-PCR and Western Blot were used to observe QHJD with effect on the TNF-a induced Hacat cell proliferation, apoptosis, Bcl-xl and Bax expression. The enzyme-linked immunosorbent assay (ELISA), RT-PCR and Western Blot was applied to observe the QHJD influence on the supernatant of TNF-a induced Hacat cell factors such as cytokines IFN-γ, IL-1β, IL-2, IL-6, IL-8 secretion and expression of p38.3.3 The CCK-8, flow cytometry, RT-PCR and Western Blot technique was used to observe the effect of Paeonol on Hacat cell proliferation, apoptosis, Bcl-xl and Bax expression. The enzyme-linked immunosorbent assay (ELISA), RT-PCR and Western Blot was applied to observe the Paeonol influence on the supernatant of Hacat cell factors such as cytokines IFN-γ, IL-1β, IL-2, IL-6, IL-8 secretion and expression of p38.3.4 The CCK-8, flow cytometry, RT-PCR and Western Blot technique was used to observe the effect of Paeonol on the TNF-a induced Hacat cell proliferation, apoptosis, Bcl-xl and Bax expression. The enzyme-linked immunosorbent assay (ELISA), RT-PCR and Western Blot was applied to observe the Paeonol influence on the supernatant of the TNF-a induced Hacat cell factors such as cytokines IFN-γ, IL-1β,IL-2, IL-6, IL-8 secretion and expression of p38.4. Result4.1 The results showed that QHJD formulae could inhibit the HaCat cells proliferation and promote apoptosis of HaCat cells, which was related to the increase expression of Bax mRNA and protein. Which could increase the expression of Bcl-xl with no statistically significance as well, which could reduce the blood detoxification of HaCaT cell cultures in IFN-γ, IL-1β,IL-2, IL-6 and IL-8 concentration in the supernatant but without statistically significant. And it could decrease the expression of p38 mRNA and protein with no statistical significance.4.2 The results showed that QHJD formulae could inhibit the TNF-a induced HaCat cells proliferation and promote HaCat cells apoptosis, which was related to the increase expression of Bax mRNA and related protein. Which could increase the expression of Bcl-xl with no statistically significance as well, which could reduce the IFN-γ, IL-1β, IL-2, IL-6 and IL-8 concentration in the supernatant of HaCaT cell but without statistically significant. And it could decrease the expression of p38 mRNA and related protein with no statistical significance.4.3 The results showed that paeonol could inhibit HaCat cells proliferation and promote HaCat cells apoptosis, which was related to the increase expression of Bax mRNA and related protein. Paeonolu could increase the expression of Bcl-xl with no statistically significance as well, which could reduce the IFN-γ, IL-1β, IL-2,1L-6 and IL-8 concentration in the supernatant of HaCaT cell but without statistically significant. And it could decrease the expression of p38 mRNA and related protein with no statistical significance.4.4 The results showed that paeonol could inhibit the TNF-α induced HaCat cells proliferation and promote HaCat cells apoptosis, which was related to the increase expression of Bax mRNA and related protein without statistical significance. Which could increase the expression of Bcl-xl with no statistically significance as well. Paeonolum could reduce the IFN-γ, IL-1β, IL-2, IL-6 and IL-8 concentration in the supernatant of TNF-α induced HaCaT cell but without statistically significant, and it could decrease the expression of p38 mRNA and related protein with no statistical significance.5. conclusionFrom the research results mentioned above, we could come to a conclusion that QHJD formulae could increase the expression of pro-apoptotic gene Bax on the one hand, thereby which could induce the keratinocyte apoptosis and inhibit abnormal keratinocyte proliferation. On the other hand, which could improve immune function to adjust abnormal keratinocyte proliferation and differentiation of cells via reducing the IFN-γ, IL-1β, IL-2, IL-6, IL-8 concentration, which could be a pathway for QHJD reducing the infiltration of inflammatory cells. Both of those factors maybe the mechanism for the treatment of psoriasis.Paeonol could induce keratinocyte apoptosis to inhibit the abnormal proliferation of keratinocytes on one hand; which could reduce the concentration of IFN-γ, IL-1β, IL-2, IL-6, IL-8 to improve the immune function for adjusting abnormal keratinocyte proliferation and differentiation of cells, for reducing the infiltration of inflammatory cells on another hand. Both of those two factors maybe the mechanism for the treatment of psoriasis.However, it is due to the complexity of traditional Chinese medicine for the various of its active ingredients but limited to the experimental conditions and. Hence the exact mechanism of QHJD on the psoriasis is in need of a lot of research work for the verification. |
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