Identifying the Effect of Nuanxin Capsules on Myocardial Injury Induced by Chronic Hypoxia via Network Pharmacology Analysis and Experimental Validation.

Nuanxin capsule (NX), an in-hospital preparation of Guangdong Provincial Hospital of Chinese Medicine, has been used in heart failure (HF) treatment for 15 years, but its mechanism and protective effect have not been investigated. This study was aimed at exploring the mechanism and protective effect of NX on HF treatment via network pharmacology analysis and experimental validation. Network pharmacology analysis predicted that NX was involved in the regulation of response to apoptotic process and hypoxia via protecting cellular damage and mitochondrial dysfunction against chronic hypoxia. Its mechanism may be involved in the regulation of the PI3K-Akt signaling pathway, HIF-1 signaling pathway, AMPK signaling pathway, and MAPK signaling pathway. Experimental validation indicated that NX was capable of improving cellular viability, restoring cellular morphology, and suppressing cellular apoptosis cellular. NX also exerted cardioprotection by inhibiting mitochondrial membrane potential injury and protecting mitochondrial respiratory and energy metabolism in a chronic hypoxia cellular model, which was consistent with the results of network pharmacology prediction. In addition, the screened active compounds of NX did have a good binding with their key targets, indicating NX may exert protective effect through multicompounds and multitargets. In conclusion, NX had a protective effect on HF through cellular and mitochondrial protection against chronic hypoxia via multicompounds, multitargets, and multipathways, and its mechanism may be involved in modulating the PI3K-Akt signaling pathway, HIF-1 signaling pathway, AMPK signaling pathway, and MAPK signaling pathway.

Nuanxin capsule enhances cardiac function by inhibiting oxidative stress-induced mitochondrial dependent apoptosis through AMPK/JNK signaling pathway.

OBJECTIVE: Oxidative stress and apoptosis play critical roles in the pathogenesis of heart failure (HF).Nuanxin capsule (NX) is a Chinese medicine that has outstanding protective effects on HF. The present study aimed to elucidate whether NX could protect HF against oxidative stress-induced apoptosis through intrinsic mitochondrial pathway. METHODS: In vivo, HF was induced by transverse aortic constriction. NX and Compound C (Comp C) were administered to C57BL/6 J mice for over a 4-week period. Cardiac function was assessed with echocardiography. In vitro, H9c2 cells were exposed to H2O2 in the presence or absence of NX and Compound C. Cell viability, cytotoxicity, reactive oxygen species (ROS) production, apoptosis, mitochondrial membrane potential (ΔΨm) and mitochondrial function by oxygen consumption rate (OCR) were detected. The expressions of cytochrome c, BAX, Bcl-2, cleaved caspase-3, AMPK and JNK were evaluated by western blotting. RESULTS: The results indicated that NX significantly improved cardiac function and enhanced the cell viability, ΔΨm and mitochondrial respiration. Also NX treatment reduced cell cytotoxicity and ROS production. Moreover, NX inhibited mitochondrial-mediated apoptosis by upregulating AMPK and downregulating JNK both in vivo and in vitro. The protective effects of NX on cardiac function by reducing oxidative stress-induced mitochondrial dependent apoptosis were reversed by Compound C treatment. CONCLUSIONS: These findings demonstrated that NX effectively improved cardiac function in TAC mice by reducing oxidative stress-induced mitochondrial dependent apoptosis by activating AMPK/JNK signaling pathway.

The Osmanthus fragrans flower phenylethanoid glycoside-rich extract: Acute and subchronic toxicity studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Osmanthus fragrans var. thunbergii (O. fragrans) flower has been consumed as folk medicine for thousands of years. O. fragrans flower extract is a well-characterized phenylethanoid glycoside-rich extract, which has been used as a natural anti-oxidant. The aim of this study was to evaluate the safety of O. fragrans flower phenylethanoid glycoside-rich extract (OFFE). MATERIALS AND METHODS: The OFFE was extracted by 80% (v/v) aqueous ethanol with 0.01% sodium isoascorbate (w/v) from the O. fragrans flower and purified on HPD300 resins. The total phenylethanoid glycosides content and individual phenylethanoid glycosides was determined by photocolorimetric method and reversed phase UPLC respectively. An acute oral toxicity study, reverse mutation test, bone marrow cell micronucleus test, and sperm abnormality test as well as a 90-day oral toxicity study were performed on experimental animals. RESULTS: The total content of phenylethanoid glycosides in OFFE was 73.4g acteoside equivalent per 100g of extract, include acteoside (52.5g per 100g of extract), salidroside (13.8g per 100g of extract), and isoacteoside (2.6g per 100g of extract) and so on. No acute lethal effect at the maximal tested OFFE dose of 10g/kg body weight (bw) in either rats or mice was observed, suggesting that OFFE can be considered nontoxic. No evidence for mutagenicity was detected in any of the three mutagenic tests. Administration at levels of 0.50, 1.00, and 2.00g/kg bw to rats for 90 days failed to induce any significant hematological, clinical, chemical, or histopathological changes. The no-observed adverse-effect-level for OFFE was >2.00g/kg bw for the study on subchronic toxicity. CONCLUSION: The results showed that consuming OFFE has no adverse effects and poses no health risk in the acute oral toxicity study, subchronic oral toxicity study, and in the micronucleus test, which may provide supportive evidence for the safety of OFFE powder that has been used in medicine as well as in functional foods, and dietary supplements.