Cryptotanshinone prevents muscle wasting in CT26-induced cancer cachexia through inhibiting STAT3 signaling pathway

Ethnopharmacological relevance: Salvia miltiorrhiza bunge (Danshen) has been extensively used to treat a wide variety of diseases including cancers. Cryptotanshinone is a major lipophilic compound extracted from the root of Danshen and has been reported to exert various pharmacological effects, however, its anti-cachectic remains unknown.

Aim of the study: The present study aims to investigate the anti-cachectic efficacy of cryptotanshinone and elucidate the underlying mechanism.

Materials and methods: Prevention of muscle wasting by cryptotanshinone in colon adenocarcinoma CT26-induced cachexia and CT26 conditioned medium (TCM)-induced myotubes were investigated. Main features of cancer cachexia were determined after cryptotanshinone administration. The therapeutic effect of cryptotanshinone on myotube atrophy was assessed by morphological observation and myotube fiber width determination. E3 ubiquitin ligases muscle RING-finger containing protein 1 (MuRF1) and muscle atrophy Fbox protein (MAFbx/Atrogin-1) expression and STAT3 activation were examined using western blot, real-time qPCR and dual-luciferase reporter gene assays both in vitro and in vivo. The myotubes were infected with lentiviruses expressing STAT3 or GFP.

Results: In CT26 tumor-bearing mice, cryptotanshinone (20 and 60 mg/kg) administration drastically prevented systemic cancer cachexia from whole body weight loss and wasting of multiple tissues including heart, fat and skeletal muscle, with a negligible effect on cancer growth at dose of 20 mg/kg cryptotanshinone administration prevented the induction of MuRF1 and MAFbx/Atrogin-1 in cachectic muscles. Moreover, cryptotanshinone (2.5-10 μM) dose-dependently reduced the elevated expression of MuRF1 and MAFbx/Atrogin-1 in C2C12 myotubes, and improved myotube atrophy. We showed that cryptotanshinone significantly suppressed the hyper-activated STAT3 in cachectic muscles and C2C12 myotubes and inhibited STAT3 transcriptional activity, but it did not repress the activation of STAT1. The inhibitory effect of cryptotanshinone on TCM-induced myotube atrophy was blocked by STAT3 overexpression.

Conclusions: These data suggest that cryptotanshinone prevents muscle wasting in cancer cachexia through STAT3 inhibition, and it may be a promising candidate drug for the treatment of cancer cachexia.

Keywords: Cancer cachexia; Cryptotanshinone; E3 ubiquitin ligases; Muscle degradation; STAT3.