| Objective To explore the effects of two Notopterygium incisum Ting ex H.T.Chang extracts on HIRI model rats and the mechanism of reducing liver injury.Methods Eighty-two 6-week-old SPF SD rats were randomly divided into four groups: Blank group(group B),simple Model group(group M),intervention group of Notopterygium incisum Ting ex H.T.Chang 70% ethanol extract(group QH-EA)and intervention group of Notopterygium incisum Ting ex H.T.Chang extract(group QH-NBA).Group m was divided into three groups(8 rats/group)according to ischemia for 30 min,ischemia for 30 min and reperfusion for 60 min,namely,group I30R0(I30R0),group I30R30(I30R30)and group I30R60(I30R60),and QH-EA group was divided into groups I30 according to the duration of ischemia and reperfusion in group M.All rats were fed adaptively for one week.After being fed adaptively,rats in group B and group M were fed with 0.9% normal saline daily,rats in group QH-EA were fed with 70% ethanol extract of Notopterygium Rhizoma daily,rats in group QH-NBA were fed with n-butanol extract of Notopterygium Rhizoma daily,and rats in all groups were fed continuously for one week.The model was made in rats only for M group,QH-EA group and QH-NBA group.The improved model method was as follows: the rats were anesthetized routinely,laparotomy was performed,the left middle lobe of the liver was blunt separated,the left middle lobe of the liver was clamped with a non-invasive arterial clamp to make the left middle lobe ischemic for 30 min,then the non-invasive arterial clamp was removed to restore the blood perfusion of the left middle lobe of the liver,and the blood perfusion of the left middle lobe of the liver was performed at the following time points of 0min,30 min and 60 min.Blood samples were collected from the inferior vena cava of rats,and the left middle lobe of liver was taken.Serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were detected by automatic biochemical analyzer,and tumor necrosis factor-a(TNF-α),interleukin-6(IL-6)and vascular endothelial growth factor-a(VEG-6)in the left middle lobe of liver were detected by double sandwich antibody method in enzyme-linked immunosorbent assay(ELISA).The pathological changes of left middle lobe of liver were observed under light microscope after hematoxylin-eosin(HE)staining.Results (1)Levels of transaminase in each group: The expression levels of serum ALT and AST of rats in group M at I30R0,I30R30 and I30R60 were higher than those in group B(P<0.05),and the highest level was at I30R60.Compared with rats in group M,blood ALT and AST levels in QH-EA group and QH-NBA group decreased at I30R0,I30R30 and I30R60(P<0.05),and the differences were statistically significant.(2)Levels of TNF-α in liver tissue: The levels of TNF-α in liver tissue of rats in group M at I30R0,I30R30 and I30R60 were higher than those in group B(P<0.05),and the highest level was found at I30R60.Compared with group M,the content of TNF-α in liver tissue of QH-EA group decreased at I30R30 and I30R60 time points(P< 0.05).The content of TNF-α in liver tissue of QH-NBA rats decreased at I30R0,I30R30 and I30R60(P<0.05),but compared with QH-NBA rats,the level of TNF-α in liver tissue of QH-EA rats decreased at I30R30(P<0.05).(3)Levels of IL-6 in liver tissue: Compared with group B,the content of IL-6 in liver tissue of rats in group M increased at three time points: I30R0,I30R30 and I30R60(P<0.05),and reached the highest level at I30R60.Compared with M group,the content of IL-6 in liver tissue of QH-EA group decreased at I30R0 and I30R60 time points(P < 0.05).In QH-NBA group,the content of IL-6 in liver tissue of rats decreased at I30R0,I30R30 and I30R60(P<0.05).At I30R0 and I30R60 time points,the level of IL-6 in liver tissue of QH-EA group was higher than that of QH-NBA group(P<0.05).(4)Levels of VEGFA in liver tissue: The content of VEGFA in liver tissue of rats in group M at three time points: I30R0,I30R30 and I30R60 was higher than that in group B(P<0.05),and the highest was at I30R60.Compared with the rats in group M,the content of VEGFA in liver tissue of QH-EA group and QH-NBA group decreased at I30R0,I30R30 and I30R60(P<0.05).(5)Observation under HE staining microscope after pathological section of the model liver lobe: the structure of hepatic lobule in group B rats is normal,and there is no dilatation and congestion in hepatic sinuses;With the progress of ischemia/reperfusion,the pathological changes of rats in group M include the existence of hepatic lobule structure,swelling and degeneration of liver cells,focal necrosis of liver cells,infiltration of lymphocytes in portal area,obvious expansion of hepatic sinuses and congestion,and the most serious injury at I30R60 time point.The pathological damage of QH-EA group and QH-NBA group was less than that of corresponding point M group.Conclusion Pretreatment with 70% ethanol extract of Notopterygium incisum Ting ex H.T.Chang and N-butanol extract of Notopterygium root can reduce transaminase level,secretion of TNF-α,IL-6 and VEGFA inflammatory factors in liver tissue of HIRI rats,alleviate inflammatory reaction and reduce liver cell damage.HE staining of pathological sections of model liver leaves showed that pretreatment with 70% ethanol extract of Notopterygium root and pretreatment with Notopterygium root n-butanol extract could reduce the degree of liver damage in HIRI rats,and it was concluded that intervention with 70% ethanol extract of Notopterygium root and Notopterygium root n-butanol extract could reduce liver cell damage in HIRI model rats and play a role in protecting liver function. |
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