Protective Effect Of GSPE On Liver Injury Induced By AAⅠ And Its Mechanism

Aristolochic acid nephropathy has been clinically confirmed,aristolochic belongs to traditional Chinese medicine,clinical application belongs to the scope of drug safety supervision of medical administration and health department.In order to explore the application of traditional veterinary medicine compatibility and detoxification theory in veterinary clinic,32 male SD rats were randomly divided into 4 groups,8 rats in each group,including blank control group,aristolochic acid group I(AA I),grape seed proanthocyanidin extract(GSPE)group,aristolochic acid group I(AA I),grape seed proanthocyanidin extract(GSPE)group,and control group Grape seed proanthocyanidins extract intervention group(AA Ⅰ + GSPE).The normal control group was given0.9% normal saline containing polyethylene glycol 400 by gavage.AA Ⅰ group was given normal saline containing polyethylene glycol 400 by gavage at 8 o’clock every day,and 10 mg / kg AA Ⅰsolution was given by gavage at 10 o’clock.In GSPE group,normal saline containing polyethylene glycol 400 was given by gavage at 8 o’clock and GSPE solution of 200 mg / kg was given by gavage at 10 o’clock.In AA Ⅰ + GSPE group,10 mg / kg AA Ⅰ solution and 200 mg / kg GSPE solution were given by gavage at 8 o’clock and 10 o’clock respectively.After 28 days of continuous administration,the rats were killed and blood was collected from abdominal vein.After the liver was taken out,the wet weight of rat liver was measured and the liver organ index was calculated.The pathological changes of rat liver tissue were observed by H.E.staining.The function of liver was detected by AST and ALT kits;Malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH PX),reduced glutathione(GSH),total antioxidant capacity(T-AOC)and other kits were used to detect the oxidation and antioxidant capacity of rat liver.TUNEL staining was used to detect the level of apoptosis of rat liver cells.QRT PCR was used to detect the relative expression levels of PI3 K and Akt m RNA in rat liver tissue,and the relative expression levels of Bax,Bcl-2,caspase-3and caspase-9 m RNA in mitochondrial apoptosis pathway.Results: the liver organ index of AA Ⅰ group and AA Ⅰ + GSPE group was significantly higher than that of normal control group(P < 0.05),and the liver organ index of GSPE group and AA Ⅰ +GSPE group was significantly lower than that of AA Ⅰ group(P < 0.05).The activities of AST and ALT in AA Ⅰ group were higher than those in normal control group(P < 0.05),while those in GSPE group and AA Ⅰ + GSPE group were lower than those in AA Ⅰ group(P < 0.05).The content of MDA in AA Ⅰ group and AA Ⅰ + GSPE group was higher than that in normal control group(P < 0.05),and the content of MDA in GSPE group and AA Ⅰ + GSPE group was lower than that in AA Ⅰ group(P< 0.05).The values of SOD,GSH Px,T-AOC and GSH in AA Ⅰ group were lower than those in normal control group(P < 0.05),while those in GSPE group and AA Ⅰ + GSPE group were higher than those in AA Ⅰ group(P < 0.05).H.E.the sections showed that the hepatocytes in AA Ⅰ group were obviously swollen,some of them were enlarged,and a few of them were turbid.The nuclei of liver were deeply stained with double nuclei.Inflammatory cell infiltration and necrosis could be seen in hepatic sinusoids.In AA Ⅰ + GSPE group,the swelling of hepatocytes was not obvious,and a few hepatocytes were deeply stained.Compared with AA Ⅰ group,the injury in AA Ⅰ + GSPE group was lighter,and the pathological changes in this group were significantly improved.TUNEL staining showed that there were fewer apoptotic positive cells in normal control group and GSPE group.Compared with normal control group and GSPE group,the number of apoptosis positive cells in AAⅠ group was significantly increased.Compared with the normal control group,the number of apoptosis positive cells in AA Ⅰ + GSPE group was more,and compared with AA Ⅰ group,the number of apoptosis positive cells decreased significantly.The relative expression levels of PI3 K,Akt and bcl-2 m RNA in AA Ⅰ group were lower than those in normal control group,while the relative expression levels of Bax,caspase-3 and caspase-9 m RNA in AA Ⅰ group were higher than those in normal control group.The relative expression levels of PI3 K,Akt and bcl-2 m RNA in GSPE group and AA Ⅰ + GSPE group were higher than those in AA Ⅰ group,while the relative expression levels of Bax,caspase-3 and caspase-9 m RNA were lower than those in AA Ⅰ group.Conclusion: GSPE has protective effect on liver injury in rats exposed to AA Ⅰ,and its mechanism is related to activation of PI3 K / Akt signaling pathway.Aristolochic acid induced excessive ROS may lead to oxidative stress,leading to cell damage.GSPE can alleviate the damage of AA Ⅰ to the liver by improving the ability of anti oxidative stress in the liver of rats.