[Cangxi Tongbi Capsules promote chondrocyte autophagy by regulating circRNA＿0008365/miR-1271/p38 MAPK pathway to inhibit development of knee osteoarthritis].

To investigate the mechanism by which Cangxi Tongbi Capsules promote chondrocyte autophagy to inhibit knee osteoarthritis(KOA) progression by regulating the circRNA＿0008365/miR-1271/p38 mitogen-activated protein kinase(MAPK) pathway. The cell and animal models of KOA were established and intervened with Cangxi Tongbi Capsules, si-circRNA＿0008365, si-NC, and Cangxi Tongbi Capsules combined with si-circRNA＿0008365. Flow cytometry and transmission electron microscopy were employed to determine the level of apoptosis and observe autophagosomes, respectively. Western blot was employed to reveal the changes in the protein levels of microtubule-associated protein light chain 3(LC3)Ⅱ/Ⅰ, Beclin-1, selective autophagy junction protein p62/sequestosome 1, collagen Ⅱ, a disintegrin and metalloproteinase with thrombospondin motifs 5(ADAMTS-5), and p38 MAPK. The mRNA levels of circRNA＿0008365, miR-1271, collagen Ⅱ, and ADAMTS-5 were determined by qRT-PCR. Hematoxylin-eosin staining was employed to reveal the pathological changes of the cartilage tissue of the knee, and enzyme-linked immunosorbent assay to measure the levels of interleukin-1ß(IL-1ß) and tumor necrosis factor-alpha(TNF-α). The chondrocytes treated with IL-1ß showed down-regulated expression of circRNA＿0008365, up-regulated expression of miR-1271 and p38 MAPK, lowered autophagy level, increased apoptosis rate, and accelerated catabolism of extracellular matrix. The intervention with Cangxi Tongbi Capsules up-regulated the expression of circRNA＿0008365, down-regulated the expression of miR-1271 and p38 MAPK, increased the autophagy level, decreased the apoptosis rate, and weakened the catabolism of extracellular matrix. However, the effect of Cangxi Tongbi Capsules was suppressed after interfering with circRNA＿0008365. The in vivo experiments showed that Cangxi Tongbi Capsules dose-dependently inhibited the p38 MAPK pathway, enhanced chondrocyte autophagy, and mitigated articular cartilage damage and inflammatory response, thereby inhibiting the progression of KOA in rats. This study indicated that Cangxi Tongbi Capsules promoted chondrocyte autophagy by regulating the circRNA＿0008365/miR-1271/p38 MAPK pathway to inhibit the development of KOA.

Cangxitongbi capsules protect the articular cartilage in the rat knee through the long non-coding RNA HOTAIR/p38MAPK pathway.

BACKGROUND: Knee osteoarthritis (KOA) is a leading cause of chronic pain and disability, and as such, it poses a significant economic burden. Traditional Chinese medicine (TCM), as well as complementary and alternative medicine, can offer safe and effective treatments for KOA. Cangxitongbi (CXTB) capsule is a Chinese patented medicine for KOA treatment and has a remarkable curative effect. This article evaluated the effects and mechanisms of CXTB in protecting joint cartilage in vivo. METHODS: The KOA model was constructed in rats using the modified Hulth method. CXTB (35 mg/kg) was administered intragastrically for 4 weeks. Hematoxylin and eosin (HE) staining of the knee articular were performed to evaluate the efficiency of CXTB. Western blot analysis, quantitative polymerase chain reaction (qPCR), and enzyme-linked immunosorbent assay (ELISA) were used to investigate the protective mechanisms of CXTB in joint cartilage. RESULTS: CXTB effectively improved the morphological structure of the cartilage and bone in the knee joint by enhancing autophagy and regulating the expression of related protease and inflammatory factors. Furthermore, CXTB downregulated the expression of the long non-coding RNA (lncRNA) Hox transcript antisense intergenic RNA (HOTAIR) and inhibited the activation of the p38MAPK pathway. Conversely, overexpression of lncRNA HOTAIR suppressed the protective effects of CXTB on the knee joint. CONCLUSIONS: CXTB capsules can protect the knee articular cartilage in rats through the lncRNA HOTAIR/p38MAPK pathway.

Cangxitongbi capsule protects articular cartilage of the knee in rats by regulating ADAMTS-5.

BACKGROUND: Knee osteoarthritis (KOA) is a disease with a high incidence in elderly patients and traditional Chinese medicine has a significant effect on the treatment of KOA. Cangxitongbi capsule (CXTB) is a traditional Chinese medicine for KOA treatment and has a remarkable curative effect. The purpose of this article is to investigate the mechanism of CXTB in protecting joint cartilage on KOA rats. METHODS: A total of 30 male Sprague-Dawley rats were randomly assigned into five groups: control group; model group; low-, mid-, and high-dose CXTB groups (17.5, 35, and 70 mg/mL). KOA models were made by modified Hulth method except the control group. After pharmacological administration for 4 weeks, knee articular cartilages were observed by hematoxylin and eosin (HE) staining and evaluated by Mankin score. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the concentration of ADAMTS-5. The peripheral blood of the rats was collected to detect content of interleukin-1 beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α) by enzyme-linked immunosorbent assay (ELISA). RESULTS: The morphological structure of cartilage in the 3 CXTB groups was significantly improved compared with the model group, and the improvement positively correlated with the drug dosage (P<0.05). Compared with the model group, the expression levels of ADAMTS-5 of the 3 CXTB groups was obviously decreased (P<0.05). Furthermore, the upstream targets of ADAMTS-5, including IL-1ß and TNF-α were down-regulated in the 3 CXTB groups (P<0.05). CONCLUSIONS: Knee joint cartilage on KOA model rats is protected by CXTB via down-regulation of ADAMTS-5. The upstream targets of ADAMTS-5, IL-1ß and TNF-α, were also down-regulated by CXTB.

Treatment of Saos-2 osteosarcoma cells with diallyl trisulfide is associated with an increase in calreticulin expression.

Diallyl trisulfide (DATS) is a natural organic sulfur compound that may be isolated from garlic and has strong anticancer activity. DATS has been demonstrated to upregulate the expression of calreticulin (CRT) in various types of human cancers, which is associated with the prognosis of cancer and its response to therapy. However, whether DATS has the same effect on human osteosarcoma cells is not known. Therefore, in the present study, Saos-2 human osteosarcoma cells were cultured with different concentrations of DATS (0, 25, 50 and 100 µmol/l) for 24 h, or with 50 µmol/l DATS for different time periods (0, 12, 24 and 36 h). Reverse transcription-quantitative polymerase chain reaction (RT-qPCR), western blotting and immunofluorescent staining were used to detect CRT mRNA and protein in the Saos-2 cells. Exposure to DATS changed the morphology and inhibited the growth of the Saos-2 cells, and its effects appeared to be concentration- and exposure time-dependent. The optimum concentration and exposure time of DATS were 50 µmol/l and 24 h, respectively. The levels of CRT mRNA and protein in the Saos-2 cells were significantly upregulated following exposure to DATS. The upregulation of CRT expression by DATS may be a mechanism underlying the ability of DATS to inhibit the growth of human osteosarcoma Saos-2 cells.