Cangxi Tongbi Capsules promote chondrocyte autophagy by regulating circRNA＿0008365/miR-1271/p38 MAPK pathway to inhibit development of knee osteoarthritis / 中国中药杂志

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-1β(IL-1β) and tumor necrosis factor-alpha(TNF-α). The chondrocytes treated with IL-1β 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.

Mechanism of Quercetin in Protecting Articular Cartilage from Osteoarthritis Based on p38 MAPK Pathway / 中国实验方剂学杂志

Objective:To investigate the protective effect of quercetin （Qu） on articular cartilage of knee osteoarthritis and its mechanism by inhibiting p38 mitogen activated protein kinase （MAPK） signaling pathway. Method:Through the network pharmacology technology，we scientifically predicted and analyzed the target factors and signal pathways of Qu in the protection of articular cartilage in patients with osteoarthritis. We selected a prediction pathway closely related to osteoarthritis and validated it by cell experiment <italic>in vitro</italic>. The best intervention concentration of the drug was selected by cell counting kit-8 （CCK-8） method. The osteoarthritis and its closely related inflammatory factors interleukin（IL）-1<italic>β</italic> and tumor necrosis factor（TNF）-<italic>α</italic> were detected by enzyme linked immunosorbent assay（ELISA）. The expression of related mRNA and protein in p38 signal pathway after Qu intervention were detected by quantitative real time polymerase chain reaction（Real-time PCR） and Western blot. Result:It was predicted that MAPK signal pathway was closely related to osteoarthritis by network pharmacology，and p38 MAPK pathway，which was most closely related to osteoarthritis，was selected for study. The results showed that 100 μmol·L^-1 Qu had the most obvious effect in decreasing the expression of related inflammatory factors，inhibited the expression of p38，phosphorylated（p）-p38，matrix metalloproteinase-13（MMP-13），A disintegrin-like and metalloproteinase with thrombospondin type-1 motifs-4（ADAMTS-4） in the pathway，and promoted the expression of CollagenⅡ. Conclusion:Qu could decrease the expression of cartilage inflammatory factors in the prevention and treatment of osteoarthritis，and the effect can be well developed by intervening and inhibiting p38 MAPK pathway related factor expression level. All the results show that Qu can decrease osteoarthritis inflammatory factors and protect articular cartilage in patients with osteoarthritis.

Protective Effect of Cangxi Tongbi Capsule on Articular Cartilage of KOA Rats Based onp38 MAPK Signaling Pathway / 中国实验方剂学杂志

Objective:To observe the therapeutic effect and mechanism of Cangxi Tongbi capsule on knee osteoarthritis (KOA) model rats. Method:Sixty 4-week-old SPF healthy male SD rats were randomly divided into blank group, model group, dimethyl sulfoxide (DMSO) group, Cangxi Tongbi capsule group, SB203580 group and Cangxi Tongbi capsule combined with SB203580 group. In addition to the normal group, the modified Hulth method was used to establish the koa model. After the model was established successfully, the Cangxi Tongbi capsule group was given 0.25 g·kg-1 Cangxi Tongbi capsule solution by gavage every day, the SB203580 group was given 0.015 g·kg-1 SB203580 solution by gavage, the Cangxi Tongbi capsule combined with SB203580 group was given a mixed solution containing 0.015 g·kg-1 SB203580 and 0.25 g·kg-1 Cangxi Tongbi capsule by gavage, the DMSO group was given 1% DMSO solution by gavage, the model group and blank group were given normal saline by gavage The stomach was killed and the material was taken after 4 weeks of drug intervention. The expression levels of interleukin-1β （IL-1β） and tumor necrosis factor-α（TNF-α）in the supernatant of peripheral blood were detected by ELISA. p38, p-p38, matrix metalloproteinase-13(MMP-13), Collagen Ⅱ mRNA and protein in p38 MAPK signal pathway were detected by Real-time fluorescence quantification PCR （Real-time PCR） and Western blot, and the localization expression of p-p38 was detected by immunohistochemistry. Result:Compared with normal group，the expression levels of p38，p-p38，MMP-13 in articular cartilage of the model group were up-regulated (P<0.01), the expression levels of CollagenⅡ was down-regulated (P<0.01). The contents of IL-1β and TNF-α in serum were significantly increased (P<0.01). Compared with model group, the expression levels of p38，p-p38，MMP-13 in articular cartilage of the Cangxi Tongbi capsule group, SB203580 group and Cangxi Tongbi capsule combined with SB203580 group were down-regulated (P<0.01), the expression levels of CollagenⅡ was up-regulated (P<0.01). The contents of IL-1β and TNF-α in serum were significantly decreased (P<0.01). Conclusion:Cangxi Tongbi capsule can effectively protect the cartilage of KOA rats, and its mechanism may be related to the targeted blocking of p38 MAPK signal pathway.

Effect of p38 MAPK Signaling Pathway in Diagnosis and Treatment of Knee Ksteoarthritis by Traditional Chinese Medicine / 中国实验方剂学杂志

Knee osteoarthritis (KOA) is a kind of joint disease characterized by progressive degeneration of articular cartilage, synovitis and pain. Its pathogenesis is not yet completely clear. Generally, it is believed that age, sex, obesity, trauma, inflammation, genetic susceptibility and other mechanical and biological factors together lead to the degradation and synthetic coupling imbalance of cartilage cells, extracellular matrix and subchondral bone. In recent years, signaling pathway has become a hot spot in the research of KOA chondrocyte proliferation and apoptosis, and the research of signal pathway in the pathogenesis and targeted therapy of KOA also began. It usually involves the expressions of cytokines, relevant genes and proteins in KOA chondrocyte. These researches mainly focus on the proliferation and apoptosis of chondrocytes, the synthesis and degradation of extracellular matrix, and the sclerosis of subchondral bone. More studies focus on p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway. Literatures show that p38 MAPK signaling pathway can regulate the proliferation and apoptosis of chondrocytes, maintain the balance of extracellular matrix metabolism, regulate the production of matrix metalloproteinases and pro-inflammatory factors, participate in the degradation of collagen and proteoglycan, and play an important regulatory role in the pathological process of KOA. Traditional Chinese medicine(TCM) therapy under the guidance of holistic concept and dialectical treatment theory has a strong pertinence and remarkable curative effect, and can control the development of the disease fundamentally. Starting with the relationship between p38 MAPK and the pathogenesis of KOA, this paper summarizes the research progress of p38 MAPK signaling pathway in the diagnosis and treatment of KOA by TCM, and provides new targets for the clinical diagnosis and treatment of KOA.