Protective Effects Of Total Flavones Of And Icariin From Epimedium Brevicornu Maxim Against Cisplatin-induced Kidney Injury

Epimedium brevicornum Maxim belongs to the genus Epimedium of the Epimedium family.It mainly effects are treating impotence,premature ejaculation,backache,leg pain,limb numbness,hemiplegia,and neurasthenia.The main active ingredient in Epimedium is total flavones of epimedium?TFE?,which have significant pharmacological effects on the immune system,skeletal system,cardiovascular system,reproductive system,and neuroendocrine system.During the processing of Epimedium,the content of flavonoids has change,and the content of Icariin will increase significantly.This is due to structural changes in the flavonoids after they are heated.Icariin is a flavonoid glycoside with high content in total flavones of epimedium?TFE?,which can improve cardiovascular and cerebrovascular function,strengthen gonads,and enhance immunity.We established models of nephrotoxicity induced by cisplatin in mice and HEK293 cells,and explored the kidneys of TFE and ICA from the aspects of renal function index,inflammation index,oxidative stress index,and apoptosis level.The role of protection,and investigated and summarized its molecular mechanism in renal protection.1.The protective effect of TFE on cisplatin-induced renal injury in mice.The mice were randomly divided into 4 groups of 8 in each group.TFE(300 and 600mg·kg^-1)+cisplatin groups were ig given TFE 300 mg·kg^-1 and 600 mg·kg^-1respectively,Normal and cisplatin group give the same dose of saline.After 1h of administration on the 7th day,cisplatin group and TFE(300 and 600 mg·kg^-1)+cisplatin group were injected cisplatin 20 mg·kg^-1 by ip to establish cisplatin kidney injury model.After the mouse eyelid blood was taken and the cervical dislocation was killed,the serum and various organ tissues were taken for the detection of each index and histopathological staining analysis.The results showed that:TFE can effectively reduce creatinine?CRE?,urea nitrogen?BUN?,malondialdehyde?MDA?levels,increase the level of glutathione transferase?GSH?,in addition TFE can also inhibit renal cell apoptosis and inflammation,reduces inflammation of the kidney tissue by inhibiting the expression of the inflammatory factor Nrf2 and reduces renal damage to kidney tissue.2.The contents of the main flavonoids of Epimedium were changed before and after the preparation of sulphur oil.Weighed 600g of epimedium accurately and divided it into 12 equal parts,each 50g and kneaded,according to the proportion prescribed by the People's Republic of China Pharmacopoeia?5:1 material ratio?.Six of the epimedium leaves were mixed with 10 g of suet oil and fried at a temperature of 90°C.Each serving was fried for 10 minutes and the epimedium leaves were fried till it was green.The remaining six epimedium leaves were directly extracted from Epimedium flavonoids.The results showed that the contents of four main flavonoids Epimedin A,Epimedin B,Epimedin C,and ICA all changed to varying degrees after sulphur oil was fired,Epimedin A decreased by 12%to 16%,Epimedin B decreased by 13%to 15%,Epimedin C decreased by 25%to 29%,and ICA increased,increasing by 40%to 52%.This result shows that after the processing of epimedium,the polyglycoside flavonoids are converted to the flavonoids of the secondary glycosides,and this change may be one of the reasons that the pharmacological activity of the epimedium has been improved after processing.3.The protective effect of ICA on cisplatin-induced nephrotoxicity.HEK293cells obtained from the ATCC cell bank were placed in a DMEM medium containing10%FBS and streptomycin and penicillin and cultured in an incubator with an environment of 95%air and 5%CO₂.When the cells are filled with 80-90%of the flask.Digest with 1 mL trypsin-EDTA solution for the next experiment.The cell viability was measured by MTT quantitative colorimetric assay,and the levels of malondialdehyde?MDA?and reduced glutathione?GSH?in cells were measured to evaluate the level of oxidative stress,the apoptosis of HEK293 cells was assessed by Hoechst 33258 staining and Western blotting,and the level of inflammation was evaluated by Western blotting.The results showed that ICA can effectively reduce the level of MDA in HEK293 cells and inhibit the decrease of GSH,and can regulate the expression levels of pro-apoptosis factor Bax,anti-apoptosis factor Bcl-2 and caspase-3/-9,antagonizes cisplatin-induced apoptosis in HEK293 cells.At the same time,using Western blotting,ICA was also found to reduce the expression of inflammatory response factors such as iNOS,NF-?B,TNF-?and IL-1?,thereby inhibiting cisplatin-induced inflammation in HEK293 cells.In summary,the results of this study clearly show that TFE and its main flavonoid component ICA can antagonize the nephrotoxicity caused by cisplatin in part by inhibiting inflammation,relieving oxidative stress and preventing apoptosis,thereby playing a role in renal protection.