Exploring Synergistic Effect of Herbal Pair of Stephania Tetrandra and Astragalus Membranaceus on Treatment of Nephrotic Syndrome by Using Integrated Metabolomics and Pharmacodynamics

Abstract

The herbal pair of Stephania tetrandra and Astragalus membranaceus (FH) is widely used in the treatment of nephrotic syndrome. Both Stephania tetrandra (FJ) and Astragalus membranaceus (HQ) can improve nephrotic syndrome. The mechanism of synergistic effect of the two drugs is not clear. To exploring synergistic effect of herbal pair of Stephania Tetrandra and Astragalus Membranaceus on treatment of nephrotic syndrome by using integrated metabolomics and pharmacodynamics. The effects of FJ, HQ, FH were evaluated by general morphological observation, hematoxylin and eosin staining, blood urea nitrogen, triglycerides, cholesterol, urinary protein, Cys C, TNF-α and IL-6. The metabolomics based on GC-MS was performed and analyzed using multivariate statistical analysis. Biomarker identification, pathway analysis, correlations between identified biomarker and efficacy indices were performed. Expressions of STAT6, p-STAT6, Bcl-2, Bax, cleaved-caspase3 were detected using western blot, respectively.The results showed that FJ, HQ, FH had obvious effects on nephrotic syndrome of rats. The pathological damage of renal tissue was improved by FJ, HQ and FH based on general morphological observation and hematoxylin and eosin staining. The content of blood urea nitrogen, triglycerides, cholesterol, urinary protein, Cys C, TNF-α，IL-13 and IL-6 was showed decreasing after regulation by FJ, HQ and FH. FH has regulated more than FJ and HQ. Eighteen metabolites (12 in serum and 6 in urine) associated with nephrotic syndrome were identified. These metabolites were mainly carbohydrates, fatty acids, organic acids and others. FH regulated more metabolites of carbohydrates and fatty acids which contribute to metabolic disorders in nephrotic syndrome than FJ and HQ. Adriamycin stimulated IL-13/STAT6 pathway and elevated the levels of several factors related to renal injury, including inflammatory response, oxidative stress and cell apoptosis, which were significantly restored by the treatment of FJ, HQ and FH in vivo. FJ, HQ and FH were all effective on treatment of nephrotic syndrome. FH was more effective than FJ and HQ for treatment of nephrotic syndrome. This study evaluated the effects of FJ, HQ and FH on nephrotic syndrome at the metabolomics level. FJ, HQ and FH significantly reduces adriamycin-induced adverse outcomes so as to prevent renal injury. Inhibition of IL-13/STAT6 pathway may be the functional mechanism under FJ, HQ and FH in experimental kidney injury. It provides a reference for the research on the herb pair and efficacy of a synergistic effect of traditional Chinese medicine.