Exploring The Effect And Mechanism Of Shaoyao Gancao Decoction Combined With Prednisone Acetate On ROS-mediated NETosis In Systemic Lupus Erythematosus

Objective The aim of this study was to investigate the effects of Shaoyao Gancao decoction and its combination with prednisone acetate on ROS-mediated neutrophil NETosis and spontaneous SLE mice,and to explore whether Shaoyao Gancao decoction combined with prednisone acetate can play a role in the treatment of systemic lupus erythematosus by participating in the process of oxidative stress and regulating ROS-mediated NETosis.Methods(1)Using ROS-mediated neutrophil NETosis as cell model,the formation of neutrophil NETs and ROS were detected by immunofluorescence to investigate the inhibitory effect of Shaoyao Gancao decoction combined with prednisone acetate on neutrophil NETs formation and ROS production.(2)Eight-week-old MRL/lpr mice(spontaneous SLE mice)were divided into four groups:model group(MRL/lpr),traditional Chinese medicine(Shaoyao Gancao decoction)group,western medicine(prednisone acetate)group and Chinese and western medicine(Shaoyao Gancao decoction combined with prednisone acetate)group,and normal control group(C57BL/6J).The physical signs of each group were recorded every day for 8 weeks,and the levels of anti-ANA antibody and anti-dsDNA antibody in serum of each group were detected by ELISA method.The content of SOD and GSH-Px in serum of mice was detected by oxidation kit,the concentration of NO in serum of mice was detected by NO kit,and the content of urinary protein was determined by urine protein kit.Results(1)Compared with the normal group,the formation of neutrophil NETs and intracellular ROS in the model group increased significantly(P<0.01),while the NETs formation and intracellular ROS production decreased significantly in the traditional Chinese medicine group and traditional Chinese and western medicine group compared with the model group(P<0.01).(2)Shaoyao Gancao decoction and its combination with prednisone acetate can effectively alleviate the proliferation and enlargement of spleen,kidney and lymph nodes of MRL/lpr mice,reduce the concentration of urinary protein and NO,and the levels of anti-ANA antibody and anti-dsDNA antibody,reduce the formation of NETs in the kidney tissue of mice,improve the renal function of mice,reduce renal injury,and play a therapeutic role in SLE.(3)Shaoyao Gancao decoction and its combination with prednisone acetate could effectively regulate the oxidative stress in SLE mice,compared with the normal control group,the contents of SOD and GSH-Px in the model group were significantly higher than those in the model group(P<0.01),and the content of GSH-Px in the serum of each treatment group was significantly higher than that in the model group(P<0.01).Conclusion Shaoyao Gancao decoction and its combination with prednisone acetate may play a good therapeutic effect on SLE by participating in the process of oxidative stress,inhibiting the production of ROS and then inhibiting the formation of NETs,regulating the process of neutrophil NETosis mediated by ROS.Shaoyao Gancao decoction can be used as a substitute for prednisone acetate to treat SLE patients with glucocorticoid resistance or patients who do not want to take hormone drugs for a long time.Shaoyao Gancao decoction can reduce the dosage of prednisone acetate and reduce the adverse reactions and side effects of hormone on patients without reducing the therapeutic effect of SLE.