Tanshinone IIA and Astragaloside IV Inhibit miR-223/JAK2/STAT1 Signalling Pathway to Alleviate Lipopolysaccharide-Induced Damage in Nucleus Pulposus Cells

Abstract

Astragaloside IV (AS IV) and tanshinone (TS IIA) are the main natural components of Salvia miltiorrhiza and Radix Astragali, respectively. The amalgam of TS IIA and AS IV has potential therapeutic value in many inflammation-related diseases. However, the aftereffect of TS IIA and AS IV for lumbar disc herniation is not clear. Although the function of miR-223 in the inflammation-related JAK/STAT pathway is unknown, it is particularly expressed in human degenerative nucleus pulposus cells. This study has investigated the efficacy of the combined application of TS IIA and AS IV in the treatment of intervertebral disc nucleus pulposus cells (NP cells) injured by lipopolysaccharide (LPS). After miR-223 inhibitor imitated NP cells, the state of the JAK family and STAT family was recognized by Western blotting (Western blot, WB) and reverse transcriptase quantitative polymerase chain reaction (qPCR). The shRNA lentivirus interference vector targeting the STAT family was constructed, and the NP cell line stably interfering with the STAT gene was established after transfection. The expression of TNF-α, IL-6, MMP-9, MMP-3, caspase-1, and caspase-3 was detected by lipopolysaccharide (WTNP cells), control virus NP cells, STAT downregulation NP cells, enzyme-linked immunosorbent assay (ELISA), Western blot, and qPCR, respectively. The cell survival rate was detected by flow cytometry and TUNEL staining reverse transcriptase-polymerase chain reaction (qPCR). NP cells were treated with TS IIA and AS IV which had been made into different concentrations, and then, the expression of miR-223, p-STAT1, and p-JAK families was detected by WB Western blotting and qPCR. MiR-223 selectively acts on JAK2/STAT1 pathway, increases the expression of TNF-α, IL-6, MMP-9, MMP-3, caspase3-1, and caspase-3, and induces apoptosis, which can be eliminated by silencing STAT1. TS IIA combined with AS IV could inhibit the expression of miR-223, p-STAT1, and p-JAK2 in NP cells, and they showed a dose-dependent tendency to p-STAT1 and p-JAK2. This study shows that miR-223 promotes the inflammatory response and induces cell injury of NP cells by acting on the JAK2/STAT1 pathway, and the combination of TS IIA and AS IV may protect NP cells by downregulating miR-223 and inhibiting the expression of JAK2 and STAT1.

Tanshinone IIA and Cryptotanshinone Counteract Inflammation by Regulating Gene and miRNA Expression in Human SGBS Adipocytes

“…The regulation of miRNAs by the two diterpenes has been previously shown in a few studies concerning different pathophysiological settings, such as cardiovascular diseases [122][123][124] or cancer [125]. Our data, therefore, strengthen and expand evidence on potential molecular mechanisms of the anti-inflammatory action of TIIA and CRY, and indicate that TIIA and CRY exert their anti-inflammatory activity in adipocytes also by controlling miRNAs expression towards an anti-inflammatory profile that may improve adipocyte dysfunction.…”

Dual specificity phosphatase 22 suppresses mesangial cell hyperproliferation, fibrosis, inflammation and the MAPK signaling pathway in diabetic nephropathy

“…The only discrepancy was that the apoptotic rate was decreased, while cleaved caspase 3 only demonstrated a decreasing trend (lacked statistical significance) in the HG-Control group compared with the LG-Control group. A possible explanation may be that cleaved caspase 3 protein expression and TUNEL-reflected cell apoptosis themselves presented a mild difference ( 32 , 33 ).…”

The potential roles of JAK/STAT signaling in the progression of osteoarthritis

“…miR-523-3p can specifically interact with ZNF667-AS1 and abolish its inhibitory effects on JAK/STAT signaling pathway ( 132 ). miR-223 can enhance LPS-induced inflammatory responses by activating the activity of JAK2/STAT1 pathway in intervertebral disc chondrocytes, increasing the expression of MMP-3, and promoting ECM degradation ( 133 ). Bioinformatics analysis has been reported that hypoxic treatment can increase the repairment of OA cartilage by promoting the proliferation and migration and suppressing the apoptosis of chondrocytes through miR-18/JAK/STAT pathway ( 134 ).…”