| Psoriasis is a common disease in the dermatology,the pathogenesis of psoriasis is complex and is thought to be related to inheritance,immunity,infection and environmental factors,Although the clinical treatment of many,but can only achieve the purpose of temporary relief,and psoriasis longer duration,recurrent and difficult to cure,seriously endangering people's physical and mental health.IL-6 is a multifunctional cytokine that secreted by keratinocytes and is highly expressed in psoriasis lesions.Calcium ion is an important second messenger involved in human coagulation,maintaining muscle excitability and other physiological functions,and keratinocyte proliferation,differentiation is also closely related.To investigate whether there is a correlation between calcium and IL-6.In this study,we investigated the effects of different concentrations of calcium on the proliferation of human keratinocyte HaCaT and the effect of IL-6 mRNA and protein in vitro,preliminary analysis of the correlation between the two factors,and then for the clinical treatment of psoriasis research and development of new drugs and drug targets for reference.Objective To study the effect of calcium on proliferation and expression of interleukin-6(IL-6)in human immortal keratinocyte colony HaCaT cells in vitro.Methods Some HaCaT cells were divided into three test groups treated with calcium at concentrations of 0.3,0.5,1mmol/L,and a control group treated with roswell park memorial institute(RPMI)1640 culture solution.After 24 hours and 48 hours cultured,methyl thiazol tetrazolium(MTT)assay was conducted to estimate the proliferation of HaCaT cells,quantitative fluorescent polymerase chain reaction(qPCR)to detect the messenger ribonucleic acid(mRNA)expression of IL-6,enzyme linked immunosorbent assay(ELISA)to measure the level of IL-6 protein in the culture supernatant of HaCaT cells,immunocytochemistry to observe the level of IL-6 protein in HaCaT cells,the data were analyzed with the Image Pro Plus 6.0 system.Statistical analysis was carried out by using independent samples t test,one-way analysis of variance and Pearson correlation analysis with the Statistical Product and Service Solutions(SPSS)19.0 software.Results Under the set concentrations,calcium showed an inducing effect on the proliferation of HaCaT cells,and the inducing effect increased with the increase of calcium concentrations.After 24 and 48 hours treatment,the proliferation rate of HaCaT cells respectively are 9.65%?0.57%,29.47%?0.73%,52.18%?1.13%(t=29.08,69.12,79.52,all P<0.01)and 10.23%?0.51%,34.17%?1.14%,57.96%?0.21%(t=35.36,51.67,87.37,all P<0.01).After 24 and 48 hours treatment,the mRNA expression and protein secretion of IL-6 were increased with the increase of calcium concentrations(24h F=133.07,121.68,P<0.01;48h F=151.17,78.64,P<0.01).Mean density of IL-6 in HaCaT cells was supported by immunocytochemistry which increased with the increase of calcium concentrations after 24 and 48 hours treatment(F=113.22,202.05,P<0.01).After 24 and 48 hours treatment,the immunocytochemistry showed that there was a positive correlation between calcium and IL-6 mRNA expression and protein secretion(24h r=0.97,0.83,P<0.01;48h r=0.98,0.91,P<0.01).Conclusion calcium can promote the proliferation of HaCaT cells under the set concentrations,and increase the mRNA and protein expression of IL-6,this maybe the reason that increased calcium concentrations lead to the proliferation of keratinocyte and aggravation of inflammation in psoriasis lesions. |
PDF Full Text Request