Radix Angelica Sinensis and Radix Astragalus ultrafiltration extract improves myocardial fibrosis in radioactive rats by regulating ROS/GSH/GPx4 axis to inhibit ferroptosis
Aim To investigate the mechanism of ROS/GSH/GPx4 axis-mediated cardiomyocyte iron death in myocardial fibrosis in radioactive rats and the intervention of Radix Angelica Sinensis and Radix As-tragalus ultrafiltration extract(RAS-RA).Methods Fifty Wistar rats were randomly divided into five groups of 10 rats in each.Except for the normal control group which was not irradiated,the rats in other groups were anaesthetised and received 40 Gy X-rays single local chest irradiation once to establish myocardial fibrosis in radioactive rats.After radiation,rats in each interven-tion group of RAS-RA were administered by gavage at doses of 0.12,0.24,and 0.48 g·kg-1,respectively,once/day for 30 consecutive days.After 30 consecutive days of administration,cardiac function of rats in each group was detected by small animal ultrasound;SOD,GSH,MDA and Fe2+levels,indicators of oxidative stress,were detected by colorimetric method in cardiac tissue of each group;fluorescence intensity of reactive oxygen species(ROS)was detected by immunofluores-cence in cardiac tissue;pathological morphology of cardiac tissue and fibrosis were observed by HE and Masson staining.And the rats in each group was used to observe the ultrastructural changes of myocardium by transmission electron microscopy.The protein expres-sion levels of NOX1,GPx4,FTH1,CO1 and α-SMA in myocardial tissue were detected by Western blot.Re-sults Compared with the blank group,the LVEF and FS values of rats in the model group were significantly reduced(P＜0.05);Fe2+level increased,MDA level increased,SOD and GSH level decreased(P＜0.05);ROS level increased;HE and Masson showed that the cardiac cell morphology was irregularly arranged,with more inflammatory cell aggregation and collagen fibre deposition.Transmission electron microscopy showed that myofilament arrangement of cardiomyocytes in the model group was irregular and loosely arranged and disordered,myofilaments were broken and dissolved,the number of damaged mitochondria was significantly increased and disordered,mitochondrial morphology was irregular,mitochondrial cristae were disorganised,mitochondria became smaller,some mitochondria were swollen,outer membrane was ruptured,and mitochon-dria with iron death-specific characteristic alterations were present;Western blot showed that the expression of α-SMA,collagen Ⅰ,and NOX1 in cardiac tissue was significantly elevated(P＜0.05),and GPX4 and FTH1 protein expression significantly decreased(P＜0.05);compared with the model group,each interven-tion group of RAS-RA significantly improved the cardi-ac function of rats,with the recovery of lipid peroxida-tion and antioxidant-related indexes,and the reduction of Fe2+level,the improvement of the structure and function of mitochondria,the reduction of the degree of fibrosis;and the myocardial tissue of α-SMA,collagenⅠ,NOX1 protein expression decreased(P＜0.05),and GPX4,FTH1 protein expression injury increased(P＜0.05).Conclusions RAS-RA inhibits myocardial fi-brosis in radioactive rats,and the mechanism may be related to inhibiting myocardial iron death by regulating the ROS/GSH/GPx4 axis.
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