Abstract: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 G 1 /S cell cycle progression in chondrocytes and the underlying mecha nism was investigated. Chondrocytes isolated from the knee o… Show more
“…The progression through each phase of the cell cycle is delicately controlled by the activity of various cyclin-dependent kinases (CDKs) and their regulatory subunits, the cyclins (6). During the G 1 phase, cyclin D1 is a key cell cycle regulatory protein that is associated with CDK4 or CDK6 in the control of cell cycle progression (7,8). Therefore, cyclin D1/CDK4 or D1/CDK6 complexes may promote cell proliferation via the promotion of the cell cycle from the G 1 to the S phase.…”
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.
“…The progression through each phase of the cell cycle is delicately controlled by the activity of various cyclin-dependent kinases (CDKs) and their regulatory subunits, the cyclins (6). During the G 1 phase, cyclin D1 is a key cell cycle regulatory protein that is associated with CDK4 or CDK6 in the control of cell cycle progression (7,8). Therefore, cyclin D1/CDK4 or D1/CDK6 complexes may promote cell proliferation via the promotion of the cell cycle from the G 1 to the S phase.…”
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.
“…Chondrocytes from the knee articular cartilage of rats were isolated, cultured and identified as previously described ( 4 ). Briefly, the chondrocytes from the knee articular cartilage of four-week-old rats were isolated using 0.2% type II collagenase (Sigma-Aldrich, St. Louis, MO, USA) in magnesium- and calcium-free phosphate buffered saline (pH 7.4) (HyClone Laboratories, Inc.) for 1 h at 37°C.…”
Section: Methodsmentioning
confidence: 99%
“…Efficient cell cycle regulation is vital for cell growth and differentiation. The family of cyclin-dependent kinases (CDKs) is one of the key regulators in the cell cycle process ( 4 , 5 ). CDKs pair with cell cycle-specific regulatory subunits known as cyclins to govern the cell cycle.…”
Bushen Zhuangjin Decoction (BZD), a well-known formulation in Traditional Chinese Medicine, has been widely used for the treatment of osteoarthritis (OA). Due to the poor intrinsic repair capacity of chondrocytes, promoting the proliferation of chondrocytes is an efficient treatment to delay the progression of cartilage degradation. The present study, therefore, focused on the effect of BZD on chondrocyte proliferation, exploring the mechanism of BZD on the inhibition of cartilage degradation. Chondrocytes isolated from the knee articular cartilage of Sprague Dawley rats were cultured and identified by type II collagen immunohistochemistry. It was found that BZD promoted chondrocyte viability in a dose- and time-dependent manner. To investigate if BZD promoted the chondrocyte viability by stimulating the cell cycle progression a flow cytometer was used, and the results showed that the percentage proportion of G0/G1 cells was significantly lower, and the percentage proportion of S cells was significantly higher, in treated cells compared with that in untreated cells. To gain insight into the mechanism underlying the effect of BZD on the cell cycle progression, the mRNA and protein expression of cyclin D1, cyclin-dependent kinase 4 (CDK4), CDK6 and p21 was measured by reverse transcription-polymerase chain reaction and western blotting, respectively. The mRNA and protein expression of cyclin D1, CDK4 and CDK6 in the BZD-treated chondrocytes was significantly upregulated, while the mRNA and protein expression of p21 was significantly downregulated, compared with that in the untreated chondrocytes. These results suggested that BZD promoted chondrocyte proliferation by accelerating G1/S transition, indicating that BZD is a potential therapeutic agent for the treatment of OA.
“…Chondrocytes were collected from the knee articular cartilage of 4-weekold Sprague-Dawley (SD) rats (Shanghai Slack Laboratory Animal Co., Shanghai, China). Chondrocytes were isolated using 0.2% type II collagenase (Sigma, St. Louis, MO, USA) in pH 7.4 magnesium-and calcium-free phosphate-buffered saline (PBS; Sigma) for 1 h at 37ÌC (20). Chondrocytes were resuspended in low-glucose DMEM (Gibco) supplemented with 10% fetal calf serum (Gibco), streptomycin (100 ”g/ml) and penicillin (100 U/ml), and seeded in monolayer at a density of 5x10 5 cells/cm 2 .…”
Abstract. We have previously reported that Tougu Xiaotong capsule (TXC) inhibits tidemark replication and cartilage degradation by regulating chondrocyte autophagy in vivo. Autophagy, a cell protective mechanism for maintaining cellular homeostasis, has been shown to be a constitutively active and protective process for chondrocyte survival. However, it remains unclear whether TXC promotes chondrocyte autophagy by regulating the autophagy-related (Atg)12/microtubuleassociated protein 1 light chain 3 (LC3) conjugation systems. Thus, in the present study, we investigated the effects of TXC on primary chondrocytes treated with cobalt chloride (CoCl 2 ). We found that CoCl 2 induced a decrease in chondrocyte viability and the autophagosome formation of chondrocytes, indicating that CoCl 2 induced autophagic death in a dose-and time-dependent manner. To determine the effects of TXC on CoCl 2 -exposed chondrocytes, we assessed cell viability by MTT assay. Our results revealed that TXC enhanced the viability of CoCl 2 -exposed chondrocytes. To gain insight into the mechanisms responsible for the enhancing effects of TXC on CoCl 2 -exposed chondrocytes, the expression of Atg genes was assessed in chondrocytes exposed to CoCl 2 and treated with or without TXC. The results revealed that the expression of beclin 1, Atg3, Atg5, Atg7, Atg10, Atg12 and LC3 II/LC3 I in the chondrocytes treated with TXC increased, compared to that in the untreated chondrocytes. In addition, ultrastructural analysis indicated that treated chondrocytes contained more autophagosomes than the untreated cells, suggesting that TXC increased the formation of autophagosomes in the chondrocytes to clear the CoCl 2 -induced autophagic death. Therefore, these data suggest that TXC is a potential therapeutic agent for the reduction of cartilage degradation that occurs in osteoarthritis.
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