Effects Of Complement Factor CD59 Deficiency On Cholestatic Liver Injury In C57BL/6 Mice

Objective:To investigate the role of complement factor CD59 in cholestatic liver injury in mouse bile duct ligation(BDL)model.Methods:(1)The anesthesia group was anesthetized by inhalation with isoflurane through a ventilator.After the laparotomy,the common bile duct was ligated and disconnected,and the abdominal cavity was sutured.Ligature and break.Mice were sacrificed on the 3rd day after surgery.(2)Determination of biochemical parameters of mouse serum liver function: The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and total bilirubin(TBil)in mouse serum were measured using a fully automatic biochemical analyzer.(3)Hematoxylin and eosin(HE)staining to observe the pathological morphology of liver tissue: HE staining of liver tissue sections of mice was used to observe the pathological morphology and structural changes of liver tissue.(4)Detection of serum C3 a and C5 a levels by ELISA: The serum complement C3 a and C5 a levels of mice were measured by ELISA,and the absorbance(A)value of each well of the microplate was detected at 450 nm on a microplate reader,and the concentration of the test sample was calculated according to the standard curve..(5)Detection of MAC expression in liver tissue by immunohistochemistry: In order to detect the amount of MAC deposition,immunohistochemical staining was performed on paraffin-embedded tissue sections.Results:(1)Compared with wild type mice,serum ALT [(730.67 ± 104.50)U / L ratio(311.33 ± 52.47)U / L,t = 8.784,P <0.001 was observed in CD59-deficient BDL mice.],AST [(877.17 ± 76.31)U / L ratio(454.83 ± 45.87)U / L,t = 11.62,P <0.001],TBil [(244.15 ± 32.22)U / L ratio(141.85 ± 41.51)U / L t = 4.769,P <0.001] increased,indicating that CD59 deficiency significantly aggravates cholestatic liver injury in mice.(2)Pathological changes of liver tissue in various mice: Severe damage to the hepatic lobules was observed in CD59-deficient BDL mice,large areas of hepatocyte necrosis were observed,and the area of stem cell necrosis in wild-type mice was significantly smaller than in CD59-deficient mice.(3)C3a [(408.30 ± 167.82)ng / ml ratio(269.48 ± 49.33)ng / ml,t = 1.587,P> 0.05] and C5 a [(284.43 ±80.88)ng / ml ratio((212.95 ± 13.53)ng / ml,t = 1.743,P> 0.05] The difference in levels was not statistically significant,indicating that the lack of CD59 did not affect the production of C3 and C5 activation products in vivo.(4)Membrane attack complex(MAC)deposition was observed to increase in liver tissue of CD59-deficient mice.Conclusion:(1)Activation of complement cascade in vivo during cholestatic hepatitis in mice.(2)Deficiency of complement factor CD59 significantly aggravates cholestatic liver injury in mice;(3)After the elimination of complement factor CD59,MAC expression in liver tissues is significantly upregulated when cholestasis hepatitis occurs in mice;(4)complement factor is eliminated The increase of cholestatic liver injury in mice after CD59 may be related to the upregulation of MAC expression.