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Xihai Li,
Wenna Lang,
Hongzhi Ye,
Fangrong Yu,
Huiting Li, Jiashou Chen,
Liangliang Cai,
Wenlie Chen,
Ruhui Lin,
Yunmei Huang,
Xianxiang Liu
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ABSTRACT: The tidemark is located between calcified and non-calcified cartilage matrices. Tidemark replication plays an important role in the pathogenesis of osteoarthrosis (OA). Autophagy, or cellular self-digestion, is an essential cellular homeostasis mechanism that was found to be deficient in osteoarthritic cartilage. This study evaluated the effects of Tougu Xiaotong capsule (TXC) on the tidemark replication and cartilage degradation, and also investigated LC3 I/II, which executes autophagy, the potential role of ULK1, an inducer of autophagy, and Beclin1, a regulator of autophagy, in the development of a papain-induced OA in rat knee joints. Using a papain-injected knee rat model, standard histological methods were used to validate our model as well as treatment with TXC or glucosamine (GlcN). After 12 weeks of treatment, the changes of cartilage structure were observed by digital radiography (DR), optical microscopy, scanning electron microscopy and transmission electron microscopy, and the LC3 I/II, ULK1 and Beclin1 levels were measured by western blotting. Cartilage degradation was evaluated by the Mankin score on paraffin-embedded sections stained with Safranin O-fast green. TXC was found to improve the arrangement of subchondral bone collagen fibers and calcium phosphate crystals, inhibit the tidemark replication and delay the cartilage degradation in the papain-induced OA. Our results also showed that LC3 I/II, ULK1 and Beclin1 levels in both the TXC+OA and GlcN+OA groups were significantly increased compared to those in the OA group. The results indicate that TXC could inhibit the tidemark replication and cartilage degradation by the regulation of chondrocyte autophagy.
International Journal of Molecular Medicine 04/2013; · 1.98 Impact Factor
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ABSTRACT: The objective of the present study was to identify chemical constituents of volatile oil from Pyrolae herba (PHVO) and evaluate the antiproliferative activity of PHVO against SW1353 human chondrosarcoma cells. The volatile oil from Pyrolae herba was prepared by hydrodistillation and characterized by gas chromatography-mass spectroscopy (GC-MS). A total of 12 components in PHVO were identified representing 81.62% of the total integrated chromatographic peaks. The major compounds were found to be n-hexadecanoic acid (29.29%), cedrol (17.08%), 6,10,14-trimethyl-2-pentadecanone (9.59%) and cis-9-octadecadienoic acid (8.23%). The antiproliferative activity of PHVO against SW1353 cells was investigated using MTT assay, flow cytometry and western blot analysis. Our results demonstrated that PHVO inhibited SW1353 cell viability in a dose- and time-dependent manner. Furthermore, PHVO treatment decreased the number of cells entering the S phase and caused a reduction in the expression of cyclin D1, cyclin-dependent kinase (CDK)4 and CDK6, whereas it caused an increase in the expression of p21. PHVO demonstrated potent antitumor activity against SW1353 cells, suggesting its potential use as a therapeutic agent in the treatment of chondrosarcoma.
International Journal of Oncology 04/2013; 42(4):1452-8. · 2.40 Impact Factor
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ABSTRACT: It has been recently shown that polysaccharides isolated from plants exhibit a number of beneficial therapeutic properties. Bauhinia championii (Benth.) Benth. has been widely used for the clinical treatment of knee osteoarthritis (OA) in China. However, the underlying molecular mechanisms of knee OA treatment have yet to be elucidated. In the present study, we investigated the effects of Bauhinia championii (Benth.) Benth. polysaccharides (BCBPs) on the proliferation and cell cycle of chondrocytes on 4-week-old male Sprague Dawley rats. Immunohistochemical staining was used to identify chondrocytes and an MTT assay was used to evaluate cell viability. Flow cytometry was used for cell cycle analysis. The mRNA and protein expression levels of cyclin D1, CDK4 and CDK6 in chondrocytes were detected using reverse transcription polymerase chain reaction (RT-PCR) and western blot analysis, respectively. The data demonstrate that BCBP treatment increased the viability of chondrocytes. In addition, BCBP treatment reduced the cell population in the G0/G1 phase, whereas the cell population was increased in the S phase. Furthermore, BCBP treatment enhanced the expression of cyclin D1, CDK4 and CDK6. These results indicate that BCBP treatment promotes cell proliferation by accelerating the G1/S transition.
Molecular Medicine Reports 03/2013; · 0.42 Impact Factor
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ABSTRACT: Achyranthes bidentata polysaccharides (ABPS) are the major bioactive constituents of Radix Achyranthes bidentata (AB), which has been widely used in traditional Chinese medicine for the treatment of osteoarthritis. However, the molecular mechanisms behind the therapeutic effect of ABPS remain unclear. In the present study, chondrocytes were isolated from Sprague-Dawley rats. The effects of ABPS on the G1/S cell cycle transition in primary chondrocytes were investigated. The chondrocytes treated with and without ABPS were analyzed and it was observed that ABPS treatment was able to enhance chondrocyte proliferation in a dose- and time-dependent manner and promote the progression of chondrocyte cell cycle proliferation via the promotion of the G1 to S phase transition. Furthermore, using RT-PCR and western blot analysis, ABPS were observed as significantly upregulating the expression of cyclin D1 and the cyclin-dependent kinases (CDKs) CDK4 and CDK6. These results suggest that ABPS are able to promote chondrocyte proliferation via the promotion of the G1/S cell cycle transition.
Molecular Medicine Reports 01/2013; · 0.42 Impact Factor
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Xihai Li,
Hongzhi Ye,
Fangrong Yu,
Liangliang Cai,
Huiting Li, Jiashou Chen,
Mingxia Wu,
Wenlie Chen,
Ruhui Lin,
Zuanfang Li,
Chunsong Zheng,
Huifeng Xu,
Guangwen Wu,
Xianxiang Liu
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ABSTRACT: Millimeter waves, high-frequency electromagnetic waves, can effectively alleviate the clinical symptoms in osteoarthritis patients, as a non-pharmaceutical and non-invasive physical therapy regimen. However, the molecular mechanisms of the therapeutic effects of millimeter wave treatment are not well understood. In the present study, the effect of millimeter waves on the G1/S cell cycle progression in chondrocytes and the underlying mechanism was investigated. Chondrocytes isolated from the knee of SD rats were cultured and identified using toluidine blue staining. The second generation chondrocytes were collected and stimulated with or without millimeter waves for 48 h. Chondrocyte viability was analyzed using the MTT assay. The cell cycle distribution of chondrocytes was analyzed by flow cytometry. mRNA and protein expression levels of cyclin D1, cyclin-dependent kinases 4 and 6 (CDK4 and CDK6) and p21 were detected using real-time PCR and western blotting, respectively. Millimeter wave stimulation was found to significantly enhance chondrocyte viability. Moreover, the percentage of chondrocytes in the G0/G1 phase was significantly decreased, whereas that in the S phase was significantly increased. In addition, following millimeter wave treatment, cyclin D1, CDK4 and CDK6 expression was significantly upregulated, whereas p21 expression was significantly downregulated. The results indicate that millimeter wave treatment promotes chondrocyte proliferation via cell cycle progression.
International Journal of Molecular Medicine 05/2012; 29(5):823-31. · 1.98 Impact Factor
