| Objective To investigate the expression and relationship between IL-17A(hereinafter referred to as IL-17) and associated cytokines in a rat mo del’s liver tissue and ileum tissue of a-naphthalene isothiocyanate induced cholestasis liver fibrosis,explore whether these cytokines take part in the formation of cholestatic hepatitis,liver fibrosis,and the damage of cholestasi s intestinal mucosal barrier.Methods 24 male healthy SD rats were randomly divided into three groups,weight 180±30g,namely the blank control group,the castor oil cont rol group and the ANIT experimental group,with 8 rats for each group.Th ere was no treatment in the blank control group.The castor oil control gro up was gavaged castor oil 5ml/kg.The ANIT experimental group was gava ged ANIT mixed with castor oil at the concentration of 2%,100mg/kg or 5ml/kg.Respectively one times per week,total five times,48 hours after the fifth times gavaging,filling the stomach to kill rats in each group,to obtai n blood from the abdominal arota.To test the serum of TBil,DBil,ALT,AS T,y-GT,ALP and TBA concentration in each group.The level of IL-17±IL- 4 and IFN-y in rats peripheral blood were detected by ELISA.HE staining was used to determine the hepatic and ileum pathological changes of the se groups.Masson staining was used to observe the changes of liver fibros is.Results 1.The morphological observation of liver tissue in HE staini ng showed that lobule and portal area were normal in the blank control g roup and the castor oil control group,no liver cell degeneration and necros is,no inflammatory cell infiltration in the portal area,no bile duct and fibr ous tissue heperplasia;ANIT experimental group can be seen that the porta 1 area expanging,many inflammatory cells (plasma cells、lymphocytes and eosinophils) infiltration,bile duct proliferation,necrotic cells exfoliated in th e bile duct cavity,bile duct cavity obstruction and fibrous tissue hyperplasi a around the bie duct;the morphological observation of ileum mucosal in HE staining showed that these three groups’ileum mucosal villus were co mplete,epithelial cells were neat,and no degeneration and necrosis.Masson’s staining showed that ANIT experimental group collagen deposition were seen around the bile duct.2.The immunohistochemical result of liver tissue showed that EL-17、I L-4 and IFN-y in the ANIT experimental group were all positive;in the bl ank control group and castor oil control group were all negative.3.In terms of these three groups’liver function,compared with blank control group (38.63±12.65) u/L and castor oil control group (60.67±11.48) u/L,ANIT experimental group (437.42±156.37) u/L was significantly increa sed in the serum of ALT;compared with blank control group (132.38±30.2 5) u/L and castor oil control group (132.67±35.90) u/L,ANIT experimenta 1 group (492.42±179.89) u/L was significantly increased in the serum ofAST;compared with blank control group (2.25±0.46) u/L and castor oil co ntrol group (2.5±0.84) u/L,ANIT experimental group (13.33±5.12) u/L wa s significantly increased in the serum of y-GT;compared with blank contro 1 group (256.25±89.83) u/L and castor oil control group (209.17±49.37) u /L,ANIT experimental group (862.79±279.12) u/L was significantly increas ed in the serum of ALP;compared with blank control group (0.52±0.12) u /L and castor oil control group (0.6±0.14) u/L,ANIT experimental group (1 47.53±23.35) u/L was significantly increased in the serum of TBil;compare d with blank control group (0.38±0.13) u/L and castor oil control group (0. 47±0.12) u/L,ANIT experimental group (117.21±20.55) u/L was significant ly increased in the serum of DBil;compared with blank control group (16. 38±5.04) u/L and castor oil control group (14.53±4.41) u/L,ANIT experi mental group (226.19±26.82) u/L was significantly increased in the serum of TBA,difference were statistically significant (all P<0.01).4.Compared with blank control group (112.25±15.82) pg/ml and castol oil control group (111.53±8.28) pg/ml,significant elevation of circulating I L-17 was demonstrated in ANIT experimental group (216.69±15.53) pg/ml, difference was statistically significant(P<0.01);compared with blank control group (24.29±8.06) pg/ml and castol oil control group (20.58±8.01) pg/ ml,ANIT experimental group of circulating IFN-y (25.53±8.45) pg/ml was a little higher,compared with blank control group (26.68±0.26) pg/ml and castol oil control group (26.74±0.25) pg/ml,ANIT experimental group of c irculating IL-4 (27.14±1.59) pg/ml was a little higher,there had no signific ant difference between these three groups (all P>0.05).5.Expression of IL-17 mRNA (3.39±1.07) in the liver tissue of ANIT experimental group was higher than blank control group (1.31±.15) and castor oil control group (1.30±0.25),difference were statistically significant (P<0.05);expression of IFN-y mRNA (31.60±14.87) in the liver tissue of ANIT experimental group was significantly increased as compared with b1 ank control group (7.35±1.07) and castor oil control group (6.87±0.62),diff erence were statistically significant(P<0.01);expression of IL-4 mRNA (4.1 9±1.31) in the liver tissue of ANIT experimental group was a little higher than blank control group (3.80±1.07) and castor oil control group (3.52± 0.77),there had no significant difference between these three groups(P>0.0 5).Expression of IL-17 mRNA (2.28±0.92) in the ileum tissue of ANIT ex perimental group was higher than blank control group (0.62±0.23) and cas tor oil control group (0.57±0.09),expression of IL-4 mRNA (0.64±0.28) in the ileum tissue of ANIT experimental group was higher than blank cont rol group (0.25±0.10) and castor oil control group (0.20±0.03),difference were statistically significant(all P<0.01);expression of IFN-y mRNA (6.18± 2.84) in the ileum tissue of ANIT experimental group was significantly in creased as compared with blank control group (2.33±0.73) and castor oil control group (2.39±0.51),difference were statistically significant(P<0.05).Conclusion 1.IL-17 and IFN-y may through exerting its pro-inflamm atory and pro-fibrogenic effect,take part in the formation of cholestatic he patitis and liver fibrosis in rat model;IL-4 may has no correlation with ch olestatic hepatitis and cholestasis liver fibrosis in rat model.2.IL-17、1IL-4 and IFN-γ might have relationship with the damage of cholestasis intestinal mucosal barrier in rat model. |
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