The Affection On Acute On Chronic Liver Failure Energy Metabolism By Oxidative Stress, Mitochondrial Permeability Transition

ObjectiveThrough the establishment of acute on chronic liver failure rat models toinvestigate the protective effect of antioxidant therapy for acute on chronic liverfailure, and learn the relationship between mitochondrial permeability transition,oxidative stress and acute on chronic liver failure energy metabolism, and based onabove disscuesions to draw evaluationa reliable indicator of oxidative stress. Toexplore the relationship beween antioxidant therapy, acute on chronic liver failuremorphology abnormal of mitochondrial, with energy metabolism, this lays thefoundation for further study.MethodsEstablished liver fibrosis in rats by intraperitoneal injecting CCL4oil solutionand then giving LPS/D-gal acute attack to establish acute on chronic liver failure(ACLF group); Using the same mean establishing acute on chronic liver failuremodal and at the beginning giving NAC transesophageal gavage to establishantioxidant therapy rat model (NAC group); Without any interruption to establishnormal rats guoup (normal group). After build rats models,we deliver all groups ofliver tissue for pathological identification. Flow cytometry liver tissue specimensfrom rat liver tissue samples gradient centrifugation liver mitochondria, ELLSA lawmitochondrial the contention of Cytc, and fluorescence microplate method todetermin the MPTP RFU; ELISA method to determin liver tissue GSH, MDA, SOD;in the levels of ROS. Specimens from rat blood samples and measures by enzymelinked immunosorbent assay plasma GSH, MDA, SOD.There were40patients from Capital Medical University Affiliated Beijing You An Hospital, which were acute (subacute) on chronic liver failure15cases, chronichepatitis B15cases, and healthy persons10cases. Observe the three groups both inblood, liver function, blood clotting like changes and oxidative stress (GSH, MDA,SOD).Results1、CCL4oil intraperitoneal injectied6weeks, rat liver tissue showed varies degrees ofliver fibrosis, manifested as increased liver volume, and pale in color, with thesurrounding tissue adhesion, the surface was small nodules and small granular, edgeobtuse, rigid texture. The light microscope views a large number of fibrous tissue,fibrous septa thickening, and no pseudolobule. After given LPS/D-gal acute attack,the light microscope views original liver injury on the basis of liver cells were chunksor sub-massive necrosis, sinusoid expansion, inflammatory cell infiltration,hypertrophy cytoplasm. Residual liver cells visible microvesicular steatosis orvacuolar degeneration.2、Three groups of rats liver and blood oxidative stress test results showed that acuteon chronic liver failure in rats at a distinct oxidative stress injury: the ACLF groupliver tissue GSH, SOD lower than normal rats and NAC rats, and there is nostatistically significant difference; liver tissue MDA is significantly higher thannormal group and NAC group, the difference is statistically significant (p <0.05).NAC liver tissue GSH, SOD levels close to normal rats, and there is no statisticallysignificant difference; liver MDA is higher than normal group, the difference isstatistically significant (p <0.05). The ACLF rats plasma GSH, SOD is significantlylower than normal group and NAC group, MDA is significantly higher than in normalrats and rats with NAC group, the difference is statistically significant (p <0.05).NAC group plasma GSH, SOD levels close to normal group, the difference is nostatistically significant.3、Detecting three groups Cyt c contention, the result is: the ACLF group0.997±0.275ng/mgprot, normal group396.6±2.063ng/mgprot and the NAC group1.406±0.656ng/mgprot, the difference is statistically significant (p <0.05). 4、 Detecting MPTP opening of three groups, the result is: the ACLF group123,152.094±53925.851RFU/mgprot,the NAC group72524.074±5168.649RFU/mgprot, and normal group48260.106±8596.065RFU/mgprot, Although thereis no statistically significant difference, but an improving trend, P value of0.068,which is closer to0.05,the course may be related to the small sample size.5、Pearson correlation analysis showed that, the MPTP openning is negativelycorrelated with mitochondrial Cytc contention, correlation coefficient r=-0.858, andATP contention is negatively correlated (r=-0.799); Mitochondrial Cytc contentionwith the amount of ATP generation is positively correlated (r=0.78).6、ACLF patients in the oxidative stress,and their plasma GSH, SOD levels aresignificantly lower, MDA levels are significantly increased: ACLF group and chronichepatitis group the blood MDA levels are elevated compared with the normalgroup,the difference is statistically significant (p <0.05); There is similar levels ofGSH between ACLF group, normal group and chronic hepatitis group, but GSHslightly lower in chronic hepatitis group. ACLF group, chronic hepatitis group andnormal group blood GSH gap significantly statistically significance (p <0.05).Conclusions1、CCL4oil solution injected6weeks can cause stable liver fibrosis modle in rats,and given LPS/D-gal acute attacks can cause acute on chronic liver failure ratmodel;2、 Affecting acute on chronic liver failure, the body is oxidative stress, while MPTPlarge number of opening, by through MPTP pore Cytc into the cytoplasm, ATPgenerated obstacles, which maybe one of the reasons for such patients in clinicalmaterial and energy metabolism;3、By exogenous supply NAC, we can supply the body reduced substances from acertain extent, improve the body’s antioxidant capacity, pathology results show thatthis antioxidant can be suppressed to a certain extent, the degree of liverfibrosisformation, delayed liver failure disease process.