Huang Lian Jie Du Tang Combined With Hyperthermia Mediates The Regulation Of JAK/STAT Signaling Pathway On The Proliferation Of HaCat Cells

Objective The proliferation of Ha Cat cells was promoted by VEGF,and the activation of JAK/STAT signaling pathway after the proliferation of Ha Cat cells stimulated by VEGF was studied.By studying the traditional Chinese medicine compound Huanglian Jiedu Decoction combined with hyperthermia intervention JAK/STAT signaling pathway,we observe the effects of cell proliferation,cycle,apoptosis and other biological characteristics after modeling.It is that Huanglian Jiedu Decoction and heat therapy promote psoriasis keratin the apoptotic mechanism of forming cells provides experimental basic research.Methods1.Using the VEGF forming cell model,CCK-8 method was used to calculate the optimal inhibitory concentration of Huanglianjiedu Decoction and Ruxolitinib.2.After the cell model was established,the use of CCK-8 was used to detect the apoptotic status by using Huanglian Jiedu Decoction,inhibitors,hyperthermia and drugs combined inhibitor group,drug combination hyperthermia group and comprehensive treatment group.The changes of cell cycle and apoptosis were detected by cytometry.RT-p CR was used to detect the mRNA expression of inflammatory factors VEGF,IL-8 and Caspase-3.3.After the cell model was established,the expression of JAK1,STAT3,and STAT1 signaling molecules were detected by quantitative PCR and Western-blot.4.After the cell model was established,the expressions of HSP70 and HSF1 related to the heat shock protein family were detected by RT-PCR and Western-blot.Results1.CCK-8 method calculated the best inhibitory concentrations of Huanglian Jiedu Decoction and Ruxolitinib were 4mg/ml and 30 um,respectively.2.After the relevant interventions,the cell cycle changes,the G1 protein degradation of the cells is slow,the proportion is significantly increased,the cells are arrested in the G1 phase;the S phase is decreased,the proportion in the S phase and the G2 phase is relatively decreased,and the G2 phase is blocked,preventing the S In the G2 phase,it is in a split-suppressed state with a pronounced apoptotic state.3.The mRNA and protein expressions of JAK,STAT3 and STAT1 were significantly higher in the model group than in the blank control group.After the intervention,Huanglian Jiedu Decoction had similar Ruxolitinib-like effects and could significantly inhibit the activation of JAK/STAT signaling pathway.PJAK1 and PSTAT3 activity was significantly inhibited and apoptosis was observed.4.The expression of VEGF,IL-8 and apoptotic factor CasPase-3 mRNA in the model group was significantly higher than that in the control group.After intervention,the mRNA expression of VEGF and IL-8 was significantly decreased;CasPase-3 The mRNA expression was significantly increased.5.The mRNA and protein expression levels of HSP70,HSF1 were significantly higher in the model group than in the blank group After intervention,they all had a decreasing trend.However,after intervention with combined inhibitors,the expression of HSP70 and HSF1 could not be decreased,but the expression was increased.Conclusion1.Huanglian Jiedu Decoction may promote apoptosis by inhibiting the activation of JAKs/STATs signaling pathway2.Huang Lian Jie Du Tang and JAKs/STATs signaling pathway inhibitor Ruxolitinib act synergistically to promote apoptosis3.Hyperthermia may interact with JAKs/STATs signaling pathway through other signal transduction pathways to promote apoptosis...