Effect of Yihuang Decoction on cGAS/STING/IRF-3 Signaling Pathway and Related Immune Factors in Subcutaneous Transplanted Tumor of Cervical Cancer Mice with Spleen Deficiency and Damp-Heat Syndrome
Objective:To investigate the effect of Yihuang Decoction on cGAS/STING/IRF-3 signal pathway and related immune factors in cervical cancer mice with spleen deficiency and damp-heat syndrome.Methods:A total of 50 SPF female BALB/c mice aged 6-8 weeks were selected and randomly divided into spleen deficiency and damp-heat syndrome model group(SDDH group,n=40)and blank group(n=10).The mouse model of spleen deficiency and damp-heat syndrome was established by hot and humid envi-ronment and high-fat and high-sugar diet in the SDDH group.The mice showed lethargy,irritability,weight loss,reduced amount of food and drinking,loose and rough hair,and feces were sometimes dry and sometimes loose with a slight increase in anal temperature,which meant the modeling was considered successful,while the blank group was fed normally.0.2 mL of 2×106/mL logarithmic growth phase HPV 16-positive human cervical cancer CaSki cell suspension was inoculated into the right axilla of 50 mice.The subcu-taneous transplanted tumor model was successful when nodules with a diameter of 5 mm or greater could be palpated after 5-7 days.According to the random number table method,the SDDH group was randomly divided into a model group and low,medium,and high-dose groups of Yihuang Decoction,with 10 mice in each group.The Yihuang decoction was given to three different dose groups through intragastric administration with concentrations of crude drug of 0.25,0.50,and 1.00 g/mL based on body mass,at a rate of 40.0 mL/kg.The model and blank groups were given the same dose of saline solution.The time of intervention was once every other day for 4 weeks.After the intervention,the researchers calculated the indexes of the thymus and spleen,tumor mass and tumor inhibi-tion rate.They also detected the level of serum interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)using the ELISA method.The HPV E6 mRNA transcription level of subcutaneously transplanted tumor tissue in the five groups was detected by branched chain DNA signal amplification.Additionally,the expression of cGAS,STING,and IRF-3 protein in transplanted tumor tissue of the five groups was determined by Western blot.Results:The model group showed a significant decrease in thymus and spleen index,as well as level of IL-6 and TNF-α compared to the blank group.The transcription level of HPV E6 mRNA was found to be significantly in-creased in the model group,while the highest tumor inhibition rate was observed in the high dose group at(35.42±4.86％)(P＜0.05);Compared with the model group,the low,middle,and high dose groups showed a significant increase in thymus and spleen index,level of IL-6,TNF-α,and the protein expression of cGAS,STING,and IRF-3.Additionally,the tumor mass and HPV E6 mRNA transcrip-tion level significantly decreased in the low,middle,and high dose groups,which was found to be proportional to the dose of Yihuang Decoction.(P＜0.05).Conclusion:Spleen deficiency and dampness-heat can promote the growth of CaSki cells and increase the ex-pression of HPV.Yihuang Decoction may enhance immune function by activating cGAS/STING/IRF-3 signaling pathway,thus inhib-iting the expression of HPV and the growth of cervical cancer in mice.
万方数据
维普
