Study On The Mechanism Of Action Of 6-hydroxydopamine On The Inflammatory Response Of Psoriasis Cell Models

ObjectiveThe effects of 6-OHDA on the proliferation,apoptosis,cell cycle and inflammatory factors of psoriasis cell model were investigated by TNF-α-induced psoriasis cell model in HaCaT cells.Methods1.TNF-α(10,25,50,100μM)was used to induce sepsis cell model in HaCaT cells.(1)CCK8 was used to detect cell proliferation activity and screen the best modeling concentration.(2)The expression of IL-1β,IL-6,IL-8 and IL-17 in psoriasis cell model was detected by RT-PCR.(3)Flow cytometry was used to detect apoptosis in psoriasis cell model.(4)Flow cytometry was used to detect cell cycle changes in psoriasis cell model.(5)Western blot was used to detect the changes of WNT signaling pathway proteins in psoriasis cell model.2.The psoriatic cell model was treated with 6-OHDA(10,25,50,100μM).(1)CCK8 was used to detect cell proliferation activity and screen the best drug concentration.(2)The expression of IL-1β,IL-6,IL-8 and IL-17 in psoriasis cell model was detected by RT-PCR.(3)Flow cytometry was used to detect apoptosis after intervention.(4)Flow cytometry was used to detect cell cycle changes after intervention.(5)Western blot was used to detect the changes of WNT signaling pathway proteins in the cell model after intervention.Results1.After HaCaT cells were treated with TNF-α(10,25,50,100 μM),(1)CCK8 results showed that compared with the normal cell group,the addition of different concentrations of TNF-α could promote cell proliferation activity(p<0.001),of which 50 μM was the most significant.(2)RT-PCR results showed that compared with the normal cell group,the expression of inflammatory factors IL-1β,IL-6,IL-8 and IL-17 in the psoriasis cell model was significantly increased(p<0.01).(3)The results of apoptosis detection showed that compared with the normal cell group,the total number of apoptosis in the psoriasis cell model was significantly reduced(p<0.001).(4)The results of cell cycle detection showed that compared with the normal cell group,the number of cells in G1 phase increased(p<0.001),the number of cells in G2 phase decreased(p<0.001),and the number of cells in S phase did not change significantly.(5)Western blot results showed that the WNT signaling pathway was activated in the psoriatic cell model compared with the normal cell group.2.After using 6-OHDA(10,25,50,100μM)to intervene in the psoriasis cell model,(1)CCK8 results showed that compared with the psoriasis cell model group,different concentrations of 6-OHDA could inhibit cell proliferation activity(p<0.001).Among them,there were too many dead cells at the concentration of 100μM,which was not conducive to subsequent experiments,because 50μM was selected as the best drug intervention concentration.(2)RT-PCR results showed that compared with the psoriasis cell model group,the expression of inflammatory factors IL-1β,IL-6,IL-8 and IL-17 in the psoriasis cell model after 6-OHDA intervention was significantly inhibited(p<0.01).(3)The results of apoptosis detection showed that compared with the psoriasis cell model group,the total number of apoptosis in the psoriasis cell model after 6-OHDA intervention was significantly increased(p<0.001).(4)The results of cell cycle detection showed that compared with the psoriasis cell model group,the number of cells in G1 phase decreased(p<0.001),and the number of cells in S phase and G2 phase increased(p<0.05)after 6-OHDA intervention.(5)Western blot results showed that compared with the psoriasis cell model group,the WNT signaling pathway in the psoriasis cell model was blocked by 6-OHDA.Conclusion6-OHDA can inhibit the proliferation activity of psoriasis cell model by blocking the activation of WNT signaling pathway,promote cell apoptosis,affect cell cycle progression,and reduce the expression of inflammatory factors IL-1β,IL-6,IL-8 and IL-17,thereby regulating the progression of psoriasis.