Study Of Effect Of Chinese Herb Medicine Of "Resolving Phlegm And Eliminating Stasis" Method On PD-L1 And IL-6/JAK1/STAT3 Signaling Pathways Of Laryngeal Carcinoma In Nude Mice

ObjectiveTo study Effect of Erchen plus Huiyanzhuyu Decoction on PD-L1,IL-6,JAK1,STAT3 of laryngeal carcinoma in nude mice.Methods1.60 nude mice were randomly divided into Phlegm-blood Stasis Syndrome group(50)and blank group(10).Feeding high-fat diet,swimming in ice water and subcutaneous injection of epinephrine hydrochloride were the method of setting up the nude mouse model of phlegm-blood stasis syndrome.During the syndrome modeling period,the general situation of nude mice was observed,and the body weight,water consumption and food intake were recorded.Detection of blood lipid and coagulation function in plasma 2 weeks later.2.Heterotopic transplantation of Hep2 laryngeal cancer cell line was used to establish a Phlegm-blood Stasis Syndrome mouse model of laryngeal cancer.The model mice were divided into control group,low,medium and high dose resolving phlegm and eliminating stasis group,and Cisplstin(DDP)group,and the administration cycle was 3 weeks.The expression level of PD-L1,IL6 mRNA in tumor tissues of mice in each group was determined by RT-PCR,and the protein expression level of PD-L1,p-JAK1,STAT3,p-STAT3 in tumor tissues of mice in each group was determined by Western blot and IHC.Results1.Compared with the blank group,the amount of diet and drinking water in the model group was less,but there was no significant difference between the two groups.In terms of weight change,after one week of modeling,compared with the model group,the blank group mice weight growth faster,the difference between the two groups was statistically significant(P<0.05).Compared with the blank group,TC,nonhdl-c and LDL-C in the model group were higher than those in the blank group.The difference between the two groups was statistically significant(P<0.05).The APTT of model group was shorter than that of blank group(P<0.05).2.① The results of RT-PCR showed that compared with the control group,the expression of PD-L1 mRNA in tumor tissue of mice in EHD low,middle and high dose group and DDP group decreased,and the expression of IL-6 mRNA in EHD low,middle and high dose group also decreased(P<0.01),but there was no significant difference in the expression of IL-6 mRNA in DDP group(P>0.05).There was no significant difference in the expression of PD-L1 and IL-6 mRNA between the EHD low and middle groups(P>0.05).The expression of PD-L1 and IL-6 mRNA in EHD high dose group was lower than that in EHD low,middle dose group and DDP group(P<0.05 or P<0.01).The expression of PD-L1 and IL-6 mRNA in EHD low and middle groups was lower than that in DDP group(P<0.01).②The results of Western blot showed that compared with the control group,PD-L1 protein expression decreased in EHD low,middle and high dose group and DDP group,STAT3 and p-STAT3 protein decreased in EHD middle,high dose group and DDP group,and p-jak1 protein expression decreased in EHD high dose group(P<0.01).PD-L1 protein expression in EHD high dose group was lower than that in EHD middle dose group and DDP group,STAT3 protein expression in EHD high dose group was lower than that in EHD low,middle dose group and DDP group(P<0.01).The expression of p-STAT3 protein in high dose EHD group was lower than that in low and middle dose EHD group(P<0.01),and there was no significant difference between high dose EHD group and DDP group(P>0.05).There was no significant difference in the expression of p-jak1 between the EHD group and DDP group(P>0.05).The expression of PD-L1 protein in DDP group was higher than that in EHD low dose group(P<0.05),and lower than that in EHD medium dose group(P<0.01).The expression of STAT3 protein in DDP group was lower than that in EHD low dose group(P<0.01),and higher than that in EHD medium dose group(P<0.05).The expression of p-STAT3 protein in DDP group was lower than that in EHD group(P<0.01 or P<0.05).③The results of IHC showed that compared with the control group,the expression of PD-L1 protein in EHD low,middle and high dose group and DDP group decreased(P<0.01).PD-L1 protein expression in high dose EHD group was lower than that in low dose EHD group(P<0.01),but there was no significant difference between high dose EHD group and middle dose EHD group(P>0.05).The expression of PD-L1 protein in middle and high dose EHD group was lower than that in DDP group(P<0.05).There was no significant difference in the expression of PD-L1 protein in EHD group compared with DDP group(P>0.05).Conclusion1.the nude mouse model of phlegm-blood stasis syndrome can be successfully set up by feeding high-fat diet,swimming in ice water and subcutaneous injection of epinephrine hydrochloride.2."Resolving phlegm and eliminating stasis" Chinese herb medicine may reduce the expression level of PD-L1 in laryngeal cancer bearing nude mice by inhibiting IL-6/JAK1/STAT3 signaling pathway.