Effect of Shenkang on renal fibrosis and activation of renal interstitial fibroblasts through the JAK2/STAT3 pathway

Abstract

Background Activation of renal fibroblasts is a critical mechanism in the process of renal fibrosis. As a commonly used herbal formula, Shenkang (SK) has been found to attenuate renal fibrosis and renal parenchyma destruction. However, the effect of SK on renal fibroblast activation in unilateral ureteral obstruction (UUO) mice and its molecular mechanism remain undetermined. The present study was performed to elucidate the effect of SK on renal fibroblast activation and renal fibrosis, as well as the potential underlying mechanism, in both NRK-49F cells and UUO mice. Methods NRK-49F cells were stimulated with 10 ng/ml TGF-β1 for 48 h. After SK treatment, the CCK-8 method was used to evaluate cell viability. Thirty-six C57BL/6 mice were randomly divided into the sham group, UUO group, angiotensin receptor blocker (ARB) group, and SK high-, moderate- and low-dose groups. UUO was induced in mice except those in the sham group. Drugs were administered 1 day later. On the 13th day, the fractional anisotropy (FA) value was determined by MRI to evaluate the degree of renal fibrosis. After 14 days, serum indexes were assessed. Hematoxylin and eosin (HE) and Sirius red staining were used to observe pathological morphology and the degree of fibrosis of the affected kidney. Western blotting and PCR were used to assess the expression of related molecules in both cells and animals at the protein and gene levels. Results Our results showed that SK reduced extracellular matrix (ECM) and α-smooth muscle actin (α-SMA) expression both in vitro and in vivo and attenuated renal fibrosis and the pathological lesion degree after UUO, suppressing JAK2/STAT3 activation. Furthermore, we found that SK regulated the JAK2/STAT3 pathway regulators peroxiredoxin 5 (Prdx5) in vitro and suppressor of cytokine signaling protein 1 (SOCS1) and SOCS3 in vivo. Conclusions These results indicated that SK inhibited fibroblast activation by regulating the JAK2/STAT3 pathway, which may be a mechanism underlying its protective action in renal fibrosis.

Effects of secretome derived from macrophages exposed to calcium oxalate crystals on renal fibroblast activation

“…During the transition of fibroblasts from quiescent state to their activated form, up-regulations of α-SMA and F-actin have been reported to promote myofibroblast contractile activity 67 , 69 . Several lines of evidence have indicated that α-SMA-associated contraction is involved in ECM organization 67 , 70 , 71 . The up-regulation of α-SMA is also important for myofibroblast focal adhesion, maturation and adhesive capability 72 , 73 .…”

Shenkang Injection and Its Three Anthraquinones Ameliorates Renal Fibrosis by Simultaneous Targeting IƙB/NF-ƙB and Keap1/Nrf2 Signaling Pathways

“…Both clinical and experimental studies have demonstrated that SKI could improve renal function in CKD. Several previous publications have highlighted that SKI retarded renal fibrosis by inhibiting levels of interleukin-6, interleukin-1β, and TNF-α ( Zhang et al, 2020 ) and modulating TGF-β1/Smad3 and JAK2/STAT3 signaling pathways ( Wu et al, 2015 ; Qin et al, 2021 ). Another study has been demonstrated that treatment with SKI could inhibit the protein expression of NF-ƙB at both mRNA and protein levels in kidney tissues of renal ischemia–reperfusion injury mice with DN induced by high-fat diet and streptozocin ( Liu, 2018 ).…”

“…Recently, clinical studies have demonstrated that SKI could improve renal function in CKD, peritoneal dialysis patients with chronic renal failure (CRF), and diabetic nephropathy ( Zhang et al, 2017 ; Song et al, 2019 ; Wang et al, 2020a ; Qin et al, 2020 ; Zou et al, 2020 ; Ma, 2021 ). A seminal publication has highlighted that SKI treatment protected against CRF and symptoms related to CKD following treatment with traditional Chinese medicine was 73.05 and 98.00%, respectively, in a clinical trial of 2200 patients ( Qin et al, 2021 ). The experimental studies revealed that SKI could improve renal function and inhibit tubulointerstitial fibrosis by anti-oxidative, anti-inflammatory, and anti-apoptotic effects in unilateral ureteral obstruction mice and rats, streptozotocin-induced mice, and renal ischemia–reperfusion injury (IRI) rats ( Liu, 2018 ; Liu et al, 2019 ; Zhang et al, 2020 ; Qin et al, 2021 ) as well as renal tubular cells or mesangial cells treated by transforming growth factor-β1 (TGF-β1) or high glucose ( Wu et al, 2015 ; Xu et al, 2016 ; Fu et al, 2019 ).…”

“…A seminal publication has highlighted that SKI treatment protected against CRF and symptoms related to CKD following treatment with traditional Chinese medicine was 73.05 and 98.00%, respectively, in a clinical trial of 2200 patients ( Qin et al, 2021 ). The experimental studies revealed that SKI could improve renal function and inhibit tubulointerstitial fibrosis by anti-oxidative, anti-inflammatory, and anti-apoptotic effects in unilateral ureteral obstruction mice and rats, streptozotocin-induced mice, and renal ischemia–reperfusion injury (IRI) rats ( Liu, 2018 ; Liu et al, 2019 ; Zhang et al, 2020 ; Qin et al, 2021 ) as well as renal tubular cells or mesangial cells treated by transforming growth factor-β1 (TGF-β1) or high glucose ( Wu et al, 2015 ; Xu et al, 2016 ; Fu et al, 2019 ). Mechanistically, several preliminary studies have revealed that SKI alleviated CKD and renal fibrosis by inhibiting pro-inflammatory cytokines such as interleukin-6, interleukin-1β, and tumor necrosis factor-α (TNF-α) expression ( Zhang et al, 2020 ) and modulating TGF-β1/Smad3 and JAK2/STAT3 signaling pathways ( Wu et al, 2015 ; Qin et al, 2021 ).…”

“…The experimental studies revealed that SKI could improve renal function and inhibit tubulointerstitial fibrosis by anti-oxidative, anti-inflammatory, and anti-apoptotic effects in unilateral ureteral obstruction mice and rats, streptozotocin-induced mice, and renal ischemia–reperfusion injury (IRI) rats ( Liu, 2018 ; Liu et al, 2019 ; Zhang et al, 2020 ; Qin et al, 2021 ) as well as renal tubular cells or mesangial cells treated by transforming growth factor-β1 (TGF-β1) or high glucose ( Wu et al, 2015 ; Xu et al, 2016 ; Fu et al, 2019 ). Mechanistically, several preliminary studies have revealed that SKI alleviated CKD and renal fibrosis by inhibiting pro-inflammatory cytokines such as interleukin-6, interleukin-1β, and tumor necrosis factor-α (TNF-α) expression ( Zhang et al, 2020 ) and modulating TGF-β1/Smad3 and JAK2/STAT3 signaling pathways ( Wu et al, 2015 ; Qin et al, 2021 ). Although SKI has been demonstrated to have anti-oxidative and anti-inflammatory effects in the treatment of CKD, little is known about its underlying oxidative stress and inflammation-associated mechanisms.…”