The Therapeutic Effects Of Orazamide On Liver Injury Induced By Isoniazid And Rifampicin In Mice And Its Possible Mechanism

Objective: To explore the therapeutic effect of orazamide on liver injury induced by isoniazid and rifampicin in mice and its possible mechanism.Method: Sixty SPF male KM mice,weighting 20 ~ 22 g,which were randomly divided into six groups(n=10)including blank group,model group,glutathione group,low-dose of orazamide group,middle-dose of orazamide group,high-dose of orazamide group.All the mice except the blank group were given isoniazid(75 mg/kg·d)+rifampicin(75 mg/kg·d)for 14 days.From the 15 th day,the glutathione group was givenglutathione 78mg/(kg·d).The low,medium and high dose groups were given orazamide 80mg/(kg·d),160mg/(kg·d),320mg/(kg·d)for14 days,while the blank group and model group were given normal saline of equal volume.28 days later,mice in each group were killed,blood was collected from eyeballs and than their livers were taken.Detection of serum liver function in mice by automatic biochemical analyzer.Histopathological changes of rat liver were observed under HE staining light microscope.A full-automatic biochemical analyzer was used to detect the activities of Superoxide Dismutase,glutathione peroxidase and Malonaldehyde in liver tissue.Enzyme-linked immunosorbent assay was used to detect the levels of tumor necrosis factor-α,interleukin-1β,cytochromeP450 enzyme and PCR was used to detect the mRNA expression levels of bile acid transporter and sodium-dependent taurocholic acid cotransporter in mouse liver homogenate.Results:(1)Compared with the blank group,the levels of serum TBIL 、 DBIL 、 ALT 、 AST 、 ALP 、 TBA in the model group were significantly increased(P<0.01).compared with the model group,the levels of TBIL,DBIL,ALT,AST,ALP and TBA could be decreased after drug treatment(P<0.01),high-dose of orazamide group had a most obvious effect among them(P<0.01).(2)The HE staining examination ofblank group was almost normal,model group showed degeneration of hepatocytes and inflammatory cell infiltration.The drug intervention groups had different degrees of improvement,and high-dose of orazamide group with glutathione group to improve the most obvious.(3)Compared with the blank group,the content of SOD 、 GSH-Px in the liver homogenate of the model group decreased significantly(P<0.01),while the content of MDA increased significantly(P<0.01),compared with the model group,the content of SOD、GSH-Px in the drug treatment group increased significantly(P<0.05),and the content of MDA decreased significantly(P<0.05),and the content of SOD and GSH-Px in the glutathione group decreased most obviously(P<0.01),the MDA in high-dose of orazamide group had a most obvious effect among them(P<0.01).(4)Compared with the blank group,the levels of TNF-α,IL-1β,CYP450 in the model group were significantly increased(P<0.01),compared with the model group,the levels of TNF-α、IL-1β、CYP450in the treatment group were significantly decreased(P<0.05),especially in the high-dose group(P<0.01).(5)Compared with the blank group,the expression levels of Bsep and Ntcp mRNA in the model group were significantly decreased(P<0.01),compared with the model group,the levels of Bsep and Ntcp mRNA in the drug treatment group weresignificantly increased(P<0.05),especially in the high-dose group(P<0.01).Conclusion: 1.Orazamide has certain therapeutic effect on liver injury caused by isoniazid combined with rifampicin.2.The therapeutic mechanism may be related to the inhibition of oxidative stress reaction,reducing the release of inflammatory factors,down-regulating the expression of CYP450 and promoting the metabolism of bile acids.