Effect Of Non-AIH Inflammation On The Initiation Of Autoimmune Hepatitis (AIH)

Objective: To investigate the role of non-AIH inflammation in the initiation of autoimmune hepatitis.Methods:(1)Mice were injected with different doses of p CYP2D6 plasmid injection via tail vein.The mice were sacrificed after 72 h and the liver tissues were harvested.The expression of h-CYP2D6 protein in liver tissues were detected by Western Blot.(2)After mice infected with Ad or CCl4,serum was taken on the 5th,10 th and 15 th day,and serum ALT level was detected by ALT kit.(3)To observe the characteristics of inflammation induced by different times of CCl4 injection.(4)After mice were infected with Ad or CCl4,the liver tissues were harvested at different time points to extract the total RNA.After reverse transcription,the expression of Tnfa,Il6 and Il1 b genes related to inflammation were detected by realtime PCR.(5)Established the animal model of autoimmune hepatitis by i.v.injecting mice tails plasmid p CYP2D6 and Ad(i.v.)or CCl4(i.p.).(6)In the Ad/p CYP2D6 group and the CCl4/p CYP2D6 group,the tail vein was continuously injected with the plasmid p CYP2D6 to observe whether the more severe autoimmune response and autoimmune hepatitis were induced.(7)The anti-CYP2D6 antibody and anti-heparin antibody titers in the serum of different treatment groups were detected by ELISA.(8)Hematoxylin-eosin(HE)staining of liver tissue sections,observe the infiltration of inflammatory cells.(9)Liver tissue sections Sirius red staining to observe the fibrosis.Results:(1)Different doses of plasmid p CYP2D6 expressed differently in the liver tissue via tail vein injection.h-CYP2D6 protein was undetectable in the liver when transfected with 1 ?g of plasmid in liver.The amount of plasmids at 10,20,50,100,200 ?g can detect h-CYP2D6 protein in the liver when transfected into the liver.From this,it can be seen that plasmid p CYP2D6 can be transfected in the liver and the amount of self-mimicking antigen expression is controllable.(2)Ad and plasmid p CYP2D6 co-treatment of mice can induce autoimmune response.The autoimmune response is related to the presence of Ad and the expression of CYP2D6.(3)After the autoimmune response induced by Ad/self-mimicking antigen in mice,continuous expression of self-mimicking antigen enhances the immune response.(4)Ad and self-mimicking antigen can induce autoimmune hepatitis,and the inflammatory response produced by Ad in liver tissue promotes autoimmune hepatitis induced by self-mimicking antigen.Autoimmune hepatitis persists after the inflammation caused by Ad disappears,and self-mimicking antigen are continuously expressed can aggravate autoimmune hepatitis.(5)CCl4 injected only once,causing acute inflammation,can return to normal in a short period of times.Four consecutive injections of CCl4 resulted in persistent inflammation similar to Ad.(6)Autoimmune responses can also be induced in mice treated with the chemical CCl4 and self-mimicking antigen.The CCl4-induced inflammatory response is controllable.CCl4-induced acute and self-mimicking antigen can not induce autoimmune responses,and only CCl4-induced persistent inflammation and selfmimicking antigen can induce autoimmune response.The autoimmune response in mice induced by CCl4 and self-mimicking antigen continues to express selfmimicking antigen that enhance the immune response.(7)Co-treatment of mice with CCl4 and plasmid p CYP2D6 could also induce autoimmune hepatitis,and the induced hepatitis persisted.Continuous expression of mimic antigens enhances autoimmune hepatitis.(8)The AIH process,whether it is induced by Ad/p CYP2D6 or CCl4/p CYP2D6,is a sustained,long-term inflammation of the liver tissue that eventually leads to hepatic fibrosis.The persistence of self-mimicking antigen aggravates fibrosis.Conclusions: Persistent non-AIH inflammation triggered by the virus or chemical factors in the liver can be an important factor in inducing AIH.Persistent non-AIH inflammation can promote self-mimicking antigen-induced autoimmune responses.When non-AIH inflammation disappears,there is still persistent typical AIH inflammation(autoimmune hepatitis)in the liver tissue and leads to liver fibrosis.The persistence of self-mimicking antigen aggravates autoimmune hepatitis and aggravates liver fibrosis.