The Effect Of JieZeNO.1on Expression Of Pattern Recognition Receptor Dectin-1in VK2/E6E7Cells

ObjectiveTo observe the effects of traditional Chinese prescription JieZe NO.1on expression of pattern recognition receptor Dectin-1and adaptive molecule Syk of human immortal ectocer-vical-vaginal epithelial cell (VK2/E6E7) stimulated by Candida albicans. To study the role of JieZe NO.1in Dectin-1/syk signaling pathway of VK2/E6E7induced by Candida albicans of human natural immune system.MethodsSection I The Expression of Dectin-1and Syk on VK2/E6E7Stimulated by Candida AlbicansThere are two groups:control (educate VK2/E6E7alone) and VK2/E6E7stimulated by heat-killed Candida albicans for6h、12h、24h、48h in vitro, respectively. The expression of Dectin-1and Syk were tested by RT-PCR.Section II The Effect of JieZeNO.1on Expression of Pattern Recognition Receptor Dectin-1and Syk on VK2/E6E7Cells Stimulated by Candida Albicans1The impact of JieZe NO.1、berberine and zymosan on the cytotoxic effects of VK2/E6E7cells.2The expression effect of Dectin-1and Syk in VK2/E6E7treated by traditional Chinese prescription JieZe NO.1. According to the result of section I, we co-cultured VK2/E6E7and heat-killed Candida albicans for24h, then divided them randomly into: model group, JieZe NO.1group、berberine group、zymosan group. Each group was treated by indicated medicine for indicated time(the concentration and time were defined according to the cytotoxic effects of VK2/E6E7cells). At the same time,control group was established. The expression of Dectin-1and Syk were tested by Western-blotting.ResultsSection I The Expression in Dectin-1and Syk Stimulated by Candida AlbicansQuantitatively test with real-time PCR showed that Dectin-1and Syk mRNA constitutively expressed in VK2/E6E7. There was up-regulated expression of Dectin-1as a result that VK2/E6E7were co-incubated with heat-killed Candida albicans for24h.(P<0.01); while the expression level was down-regulated after48h co-incubation. The expression trend of downstream adaptive molecule Syk was same as Dectin-1.Section II The Effect of JieZeNO.1on Expression of Pattern Recognition Receptor Dectin-1and Syk in VK2/E6E7Cells Stimulated by Candida Albicans.1The impact of JieZe NO.1、berberine hydrochloride and zymosan on the cytotoxic effects of VK2/E6E7cells10.792mg/ml was the IC10value of JieZe NO.1on VK2/E6E7for24h;3.626μmol/L was the IC10value of berberine hydrochlorideon VK2/E6E7for12h;51.978mg/ml was the IC10value of zymosan on VK2/E6E7for12h.2The expression effect of Dectin-1and Syk inVK2/E6E7treated by traditional Chinese prescription JieZe NO.1 Compared model group with control group, the expression of Dectin-1in VK2/E6E7increased obviously, which was statistically significant (P<0.01); Compared JieZe NO.1group、berberine group、zymosan group with model group, the decreased expression of Dectin-1in VK2/E6E7was statistically significant (P<0.01); while there was no statistical differences among JieZe NO.1group、berberine group and zymosan group.(P>0.05)The expression trend of downstream adaptive molecule Syk was same as Dectin-1. That was, compared with Con,there was statistical differences about the increased expression of Syk in model group (P<0.01). The expression of Syk in JieZe NO.1group berberine group、zymosan group was fewer than in control group, which was statistically significant(P<0.01); while there was no statistical differences among JZ1group、berberine group and Zym group.(P>0.05).Conclusion1. There is constitutive expression of Dectin-1and Syk in VK2/E6E7.2. The expression of pattern recognition receptor Dectin-1is up-regulated in VK2/E6E7stimulated by heat-killed Candida albicans., with the expression of Syk was on a rising. It suggest that Candida albicans could be recognized by the Dectin-1on surface of VK2/E6E7, via Syk-dependent pathway mediating innate immune responses. Dectin-1may play a partial role in the recognition of Candida aibicans and startup host defense mechanism of Candida albicans infection3. JieZe NO.1down-regulates the expression of pattern recognition receptor Dectin-1and adaptive molecule Syk in VK2/E6E7infected by Candida albicans, consequently give rise to reducing downstream proinflammatory cytokines, and limit local inflammatory reactions.