| Objective: To compare the expression of key molecular proteins of c GASSTING-TBK1-IRF3 signal pathway in peripheral blood mononuclear cells(PBMCs)between patients with immune tolerance stage and immune clearance stage of HBV infection and healthy controls,and the effect of Bu Shen Jie Du Formula on the expression of key molecular proteins of PBMCs c GAS-STING-TBK1-IRF3 signal pathway and IFN-α and IFN-β in patients with HBV infection in immune tolerance stage and immune clearance stage,so as to further explain whether Bu Shen Jie Du Formula can restore the immune function of patients with HBV infection and provide further basis for clinical application of Bu Shen Jie Du Formula in the treatment of patients with chronic HBV infection.Methods: 20 patients with chronic HBV infection were collected from March 2021 to September 2021 in the Department of Hepatology,the first affiliated Hospital of Hunan University of traditional Chinese Medicine,including 10 patients with immune tolerance and 10 patients with immune clearance.After informed consent of patients,peripheral venous blood 30 ml was collected In addition,10 healthy people without HBV infection were collected.Peripheral blood samples from 10 ml were collected,and PBMCs were isolated,Western Blot method was used to de tect the expression of PBMCs c GAS-STING-TBK1-IRF3 signal pathway molecular proteins in immune tolerance stage,immune clearance stage of HBV infection and healthy subjects.In addition,after PBMCs were isolated from peripheral venous blood 20 ml,the same volume of Bu Shen Jie Du Formula plasma and blank plasma were intervened for 48 h respectively.After 48 h,PBMCs was collected for Western Blot and ELISA to detect the protein expression level of c GAS,STING,TBK1,IRF3 and the expression level of IFN-α and IFN-β between groups.SPSS26.0 was used for statistical analysis of the data.Results: 1.The expression levels of PBMCs c GAS protein in patients with immune tolerance stage,immune clearance stage of HBV infection and healthy persons were 0.36±0.09,0.16±0.13,0.36±0.09 respectively,The expression level of PBMCs c GAS protein in patients with immune clearance stage of HBV infection was significantly lower than that in healthy subjects and patients with immune tolerance stage of HBV infection and the difference was statistically significant(P<0.05);However,the expression level of PBMCs c GAS protein in patients with immune tolerance was not significantly different from that in healthy subjects(P>0.05).The expression levels of PBMCs STING protein in the three groups were 0.39±0.26,0.16±0.12,0.46±0.18 respectively.The expression level of PBMCs STING protein in patients with immune tolerance and clearance was lower than that in healthy subjects,and the expression level of PBMCs STING protein in patients with immune clearance was significantly lower than that in healthy subjects and the difference was statistically significant(P<0.05).The expression levels of PBMCs TBK1 protein in the three groups were 0.60 ±0.21,0.34±0.20,0.49±0.17 respectively.The expression of PBMCs TBK1 protein in patients with immune tolerance of HBV infection was the highest,followed by healthy people.The expression level of PBMCs TBK1 protein in patients with immune clearance was significantly lower than that in patients with immune tolerance and the difference was statistically significant(P<0.05).The expression levels of IRF3 protein in the three groups were 0.86±0.30,0.45±0.37,0.58±0.40 respectively.The expression level of PBMCs IRF3 protein in patients with immune tolerance stage of HBV infection was the highest,followed by healthy people.The expression level of PBMCs IRF3 protein in patients with immune clearance stage of HBV infection was significantly lower than that in patients with immune tolerance stage and the difference was statistically significant(P<0.05)..2.After intervention with Bu Shen Jie Du Formula plasma and blank plasma,the expression levels of PBMCs c GAS protein in immune tolerance stage of HBV infection were 0.13 ±0.11,0.15±0.11 respectively,the expression levels of PBMCs STING protein were 0.28±0.21,0.28±0.23 respectively,the expression levels of PBMCs TBK1 protein were 0.39 ±0.24,0.37 ±0.23 respectively,and the expression level of IRF3 protein was 0.11±0.07,0.13±0.10,respectively There was no significant difference in the above indexes between the two groups(P>0.05).The expression levels of IFN-α in PBMCs culture supernatants of the above two groups were 72.59±19.43pg/ml and 49.52±14.59 pg/ml respectively,and the expression levels of IFN-β were 1120.85±213.29pg/ml and 684.49±218.74pg/ml respectively.The expression levels of IFN-α and IFN-β in Bushen jiedu intervention group were significantly higher than those in blank plasma control group(P<0.05).3.After intervention with Bu Shen Jie Du Formula plasma and blank plasma,The expression levels of PBMCs c GAS protein in immune clearance stage of HBV infection were 0.11±0.05,0.17±0.15 respectively,the expression levels of PBMCs STING protein were 0.26±0.22,0.23±0.16 respectively,the expression levels of PBMCs TBK1 protein were 0.13±0.11,0.13±0.10 respectively,and the expression levels of PBMCs IRF3 protein were 0.14±0.12,0.08±0.06 respectively.There was no significant difference in the above indexes between the two groups(P>0.05).The expression levels of IFN-α in the culture supernatant of PBMCs in the above two groups were 83.88±11.80pg/ml and 56.14±7.73 pg/ml respectively and the expression levels of IFN-β were 1100.56±193.10pg/ml,860.81±113.27pg/ml respectively.The expression levels of IFN-α and IFN-β in Bushen jiedu intervention group were significantly higher than those in blank plasma control group(P<0.05).Conclusion: 1.There was a dynamic decrease in the expression of key molecular proteins of PBMCs c GAS/STING signal pathway from immun e tolerance to immune clearance in patients with chronic HBV infection.2.Bu Shen Jie Du Formula had no significant effect on the expression of PBMCs c GAS,STING,IRF3 and TBK1 in PBMCs of patients with immune tolerance / clearance of HBV infection,but could increase the expression of type I interferon in PBMC of patients with immune tolerance and clearance of HBV infection.It is suggested that Bu Shen Jie Du Formula may promote the expression of type I interferon in other ways,so as to further improve the immune function of patients with chronic HBV infection. |
PDF Full Text Request