Shen Shuai Ⅱ Recipe Attenuates Renal Fibrosis in Chronic Kidney Disease by Improving Hypoxia-induced the Imbalance of Mitochondrial Dynamics Via PGC-1α Activation

Background: Shen Shuai Ⅱ Recipe (SSR) is an effective Chinese herbal formula for the treatment of patients with chronic kidney disease (CKD) in the clinic and significantly improves 5/6 ablation and infarction (A/I) surgery-induced renal interstitial fibrosis (RIF) and intrarenal hypoxia in rats. However, the underlying molecular mechanisms need further elucidation.

Purpose: This study aims to investigate the renoprotective mechanisms of SSR in vivo and in vitro.

Methods: CKD model was induced in rats with 5/6 (A/I) surgery. 4 weeks later, rats were treated with vehicle or SSR or Fenofibrate by daily gavage. In vitro, HK2 cells exposed to hypoxia (1% O) were treated with SSR in the presence or absence of 100 μM MitoTEMPO or 10 μM Mitochondrial Fusion Promoter M1. The effects of SSR on RIF, mitochondrial dynamics, oxidative metabolism, and mitochondrial ROS (mtROS) were determined by immunoblotting, colorimetric, and fluorometric assays according to the experimental protocols. Furthermore, to explore the mechanisms of SSR against RIF, HK2 cells of PGC-1α or MFN2 knockdown under hypoxic stimulation were treated with 400 μg/ml of SSR and (or) 1 μM of ZLN005.

Results: The results showed that treatment with SSR significantly improved mitochondrial morphology and function, up-regulated the expression of PGC-1α protein, and inhibited the production of mtROS in 5/6 (A/I) kidneys and hypoxia-treated HK2 cells, which may be closely correlated with its anti-RIF effect. In addition, compared to wild-type HK2 cells, the roles of SSR in improving mitochondrial dynamics and energy metabolism were greatly diminished in HK2 cells of PGC-1α knockdown under hypoxic exposure. More importantly, compared to ZLN005 or SSR combined with ZLN005 treatment, MFN2-deficient HK2 cells exhibited the increased protein levels of FN, α-SMA, TGF-β1 and cleaved IL-1β in response to hypoxic stimulation.

Conclusion: SSR exerted the renoprotective effects by improving mitochondrial dynamics under hypoxic condition via PGC-1α activation.