Effects And Mechanisms Of Kochia Scoparia Saponins On M5-induced Inflammation And Proliferation Of HaCaT Cell

Objective: To study the effect of Radix saponins on M5-induced inflammation and proliferation of HaCaT cells,and to explore the possible mechanism of Radix saponins in the treatment of psoriasis.Methods:(1)MTT was used to determine the effect of saponin on the activity of HaCaT cells,and the optimal concentration and time of saponin were selected.(2)M5(IL-17 A,IL-22,oncostatin M,IL-1α,and TNF-α)cytokines cultured HaCaT cells to simulate the in vivo environment of psoriatic cells and build psoriatic cell models in vitro.(3)CCK-8 was used to detect the effect of psoriasis cell models construction and the effect of radix saponins on the cell viability of psoriasis cell models.(4)Wb was used to detect the effects of saponins on the expression of β-Catenin,c-Myc and VEGF in psoriasis cell models.(5)ELISA and qPCR were used to detect the effects of saponin on the secretion and mRNA expression of IL-1β,IL-8,IL-18 and IL-6 in psoriasis cell models.(6)Wb was used to detect the effect of saponins on the expression of TLR4 and NF-κB p65 in psoriasis cell models.Results:(1)Based on the results of MTT,compared with the Control group,the activity of HaCaT cells treated with 5,10,25,50 and 100 μmol/L saponins was significantly decreased at24 h,and after 48 h of cell growth,The inhibitory effect of 50 μmol/L and 100 μmol/L saponins on HaCaT cell proliferation was decreased,and the difference was statistically significant.(2)Based on the results of CCK-8 and inverted microscope,compared with the Control group,the cell vitality of M5 group was higher,the cell vitality of Momordin Ic group was lower,compared with the M5 group,the cell vitality of M5+Momordin Ic group was lower,the difference was statistically significant.(3)Based on the results of Wb,compared with the Control group,the protein expressions of β-catenin,c-Myc and VEGF in the M5 group were increased,while the protein expressions of β-catenin,c-Myc and VEGF in the Momordin Ic group were decreased;Compared with M5 group,the protein expressions of β-catenin,c-Myc and VEGF in M5+Momordin Ic group decreased,and the differences were statistically significant.(4)Based on the results of ELISA and qPCR,Compared with the Control group,the mRNA expression and secretion of IL-1β,IL-8,IL-18 and IL-6 in the M5 group were increased,while that of IL-1β,IL-8,IL-18 and IL-6 in the Momordin Ic group were decreased.Compared with the M5 group,the mRNA expression and secretion of IL-1β,IL-8,IL-18 and IL-6 in M5+Momordin Ic group were decreased,and the differences were statistically significant.(5)Based on the results of Wb,compared with the Control group,the expressions of TLR4 and NF-κB p65 in the M5 group were significantly up-regulated,while those in the Momordin Ic group were significantly reduced.Compared with the M5 group,the expressions of TLR4 and NF-κB p65 in M5+Momordin Ic group were significantly decreased,and the differences were statistically significant.Conclusion: The inflammatory and proliferation state of HaCaT cells were up-regulated by M5 cytokines.It is suggested that radix saponins may act on the downstream inflammatory factor target genes by activating the TLR4-mediated NF-κB pathway to reduce the inflammatory response of psoriasis and inhibit the wnt/β-actenin pathway to reduce cell viability for psoriasis.