Duhuo Jisheng Decoction promotes chondrocyte proliferation through accelerated G1/S transition in osteoarthritis

Abstract

Abstract. Duhuo Jisheng Decoction (DHJSD), a well known traditional Chinese folk medicine, is used for eliminating stagnation, removing blood stasis, promoting blood circulation and alleviating pain; it is commonly used for the treatment of various diseases, including osteoarthritis (OA). However, the molecular mechanisms behind the therapeutic effects of OA remain unclear. In the present study, the effects of DHJSD on the morphology of articular cartilage and the G1/S cell cycle progression in chondrocytes, as well as the underlying mechanisms, were investigated. A total of 27 two-month-old male Sprague Dawley rats were randomly divided into 3 groups: the control group (no papain-induced OA; received an equivalent amount of saline only), the model group (papain-induced OA; received an equivalent amount of saline only) and the DHJSD group [papain-induced OA; received a clinical oral dose of DHJSD (9.3 g/kg/day)]. After 8 consecutive weeks of treatment, the morphological changes in articular cartilage were observed under an optical microscope and by transmission electron microscopy (TEM) and the mRNA and protein expression levels of cyclin D1, CDK4, CDK6, retinoblastoma protein (Rb) and p16 were measured by RT-PCR and immunohistochemistry, respectively. Treatment with DHJSD significantly improved the arrangement of collagen fibers in the articular cartilage, as well as its structure and reduced cell degeneration compared with the model group. The mRNA and protein expression levels of cyclin D1, CDK4, CDK6 and Rb in the DHJSD-treated group were significantly increased compared with those in the model group, whereas p16 expression was significantly downregulated. Taken together, these results indicate that DHJSD treatment promotes chondrocyte proliferation by promoting the G1/S checkpoint transition in the cell cycle and by upregulating the expression of cyclin D1, CDK4, CDK6 and Rb and downregulating the expression of p16 and this may, in part, explain its clinical efficacy in the treatment of osteoarthritis.

Duhuo Jisheng Decoction-containing serum promotes proliferation of interleukin-1β-induced chondrocytes through the p16-cyclin D1/CDK4-Rb pathway

“…It has been widely used for treating OA (21), and a previous study indicated that DHJSD contains drug-and lead-like compounds with potential synergy and polypharmacology against OA (22). An in vivo study demonstrated that treatment with DHJSD promotes the progression of chondrocytes from G 1 to S phase (23), and this may be one of the mechanisms underlying its use in OA. In addition, previous studies have reported that chondrocytes treated with IL-1β produce a particularly effective cell model of the mechanisms involved in degenerative arthropathies (24).…”

Duhuo Jisheng Decoction inhibits SDF-1-induced inflammation and matrix degradation in human degenerative nucleus pulposus cells in vitro through the CXCR4/NF-κB pathway

“…As shown in Figure 7, DHJSD has an overall effect to eliminate "fenghan-shi", respectively, and thus to relieve pain of "Bi Zheng" through promoting "Gan" and "Shen" function and therefore strengthen the bones and tendons [30] . It also improves circulation and remove stasis of "Qi" and "Xue", reduces tissue swelling and relieves pain [20,22,23,26,32,39,40] . Some pharmacological studies have demonstrated that DHJSD may have effects of analgesia, immune function regulation, blood vessel dilation, anti-inflammation, and antiplatelet aggregation [20,22,23,26,32,39,40] .…”

“…It also improves circulation and remove stasis of "Qi" and "Xue", reduces tissue swelling and relieves pain [20,22,23,26,32,39,40] . Some pharmacological studies have demonstrated that DHJSD may have effects of analgesia, immune function regulation, blood vessel dilation, anti-inflammation, and antiplatelet aggregation [20,22,23,26,32,39,40] . In a study of the aging-related osteoarthritis, another common bone degenerative disease, Chen et al found that DHJSD may inhibit VEGF and HIF-1α to retard cartilage degradation in a rabbit model of osteoarthritis [53] .…”

“…DHJSD was specifically developed to orchestrate the therapeutic actions to target at the primary (Jun) and secondary (Chen and Zuo) causatives [31] and has an overall effect to eliminate "feng-han-shi" and thus relieves pain of "Bi Zheng" through promoting "Gan" and "Shen" function and reinforcing the bones and tendons [30]. It also improves circulation and removes stasis of "Qi" and "Xue", reduces tissue swelling, and relieves pain [20,22,23,26,32,39,40]. Duhuo (contains osthole) and jisheng target the major etiological problems as "Jun" (or chief) components by eliminating "feng-han-shi" [21].…”

“…The main function of SDF-1 is to guide cell migration and to further stimulate the chemotaxis of inflammatory cells and induce cell surface to produce integrins. Our previous studies have shown that the SDF-1 expression and apoptosis in degeneration of human (Poria Cocos), Chuanxiong (Rhizoma Chuanxiong), Danggui (Radix Angelicae Sinensis), Niuxi (Radix Achyranthis Bidentatae), Dihuang (Radix Rehmanniae Preparata), Baishao (Radix Paeoniae Alba), Duzhong (Cortex Eucommiae Ulmoidis), Renshen (Panax ginseng) and Gancao (Radix Glycyrrhizae) [20,22,23,26,39,40] , were mixed and soaked in distilled water, rotary evaporator for rotary steam and the solution was filtered and concentrated [41] . After boiled for 30 min twice the DHJSD solution was filtered (filter 0.22 µm) and then dissolved in DMEM/F12 medium containing 15% fetal bovine serum at a final concentration of 10 mg/mL.…”