ABSTRACT
BACKGROUND:Tougu Xiaotong Capsule (TGXTC) is a clinical prescription for the treatment of osteoarthritis;however, its mechanism has not been ful y elucidated. Urokinase-type plasminogen activator (uPA) system participating in the degradation of the extracel ular matrix of articular cartilage and hyperplasia of joint synovium plays an important role in the pathological process of osteoarthritis. OBJECTIVE:To investigate the effects of TGXTC medicated serum on the expression of uPA, uPA receptor (uPAR), plasminogen activator inhibitors (PAIs), matrix metal oproteinase-3 (MMP-3), interleukin-1 beta (IL-1β) and tumor necrosis factor-α(TNF-α) in osteoarthritis synovial cel s of rats and to discuss the mechanism by TGXTC medicated serum prevents and cures osteoarthritis. METHODS:Rat models with knee osteoarthritis were established by injecting 4%papain into the knee joint cavity. Primary synoviocytes and osteoarthritis synoviocytes were cultured with col agenase digestion method. The cultured synoviocytes were divided into normal group, model group and TGXTC group. The western blot method was adopted to detect uPA, uPAR, PAI, MMP-3, IL-1βand TNF-αprotein expression of synoviocytes after acting by TGXTC medicated serum for 72 hours. RESULTS AND CONCLUSION:The expression of uPA, uPAR, MMP-3, IL-1βand TNF-αwere decreased, while PAI was increased in the TGXTC group, and there were significant differences when compared with model group. In a word, TGXTC can significantly inhibit the expression of uPA, uPAR, MMP-3, IL-1β, TNF-α, and improve PAI expression in synoviocytes, which may partly explain the mechanism of the treatment of Tougu Xiaotong Capsule on osteoarthritis.
ABSTRACT
BACKGROUND:Tougu Xiaotong capsule is the clinical prescription for the treatment of osteoarthritis, however, its mechanism has not been fuly elucidated. Urokinase type plasminogen activator system which participated in the degradation of the extracelular matrix of articular cartilage and hyperplasia of joint synovium plays an important role in the pathological process of osteoarthritis. OBJECTIVE: To determine the effect ofTougu Xiaotong capsule on urokinase-type plasminogen activator system in knee cartilage tissues of knee osteoarthritis rats. METHODS: Of 144 Sprague-Dawley rats, 120 rats were randomly made into models of knee osteoarthritisvia intra-articular injection of papain, and randomly assigned to model group,Zhuanggu Guanjie Wan group [1.2 g/(kg?d)], low-doseTougu Xiaotong capsule group [0.092 g/(kg?d)], moderate-doseTougu Xiaotong capsule group [0.184 g/(kg?d)] and high-doseTougu Xiaotong capsule group [0.368 g/(kg?d)]. Each group contained 24 rats. Every 2 weeks was considered as a course, with a 2-day interval, totaly 4 courses. The remaining 24 normal rats were included in the blank group. After every two courses, a batch of experimental animals was sacrificed. The pathological changes were observed folowing staining with hematoxylin and eosin. The positive cels of urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor and plasminogen activator inhibitor were measured by immunohistochemistry. The protein levels of urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor and plasminogen activator inhibitor were measured by western blot assay. RESULTS AND CONCLUSION:Mankin’s score was significantly lower in theTougu Xiaotong capsule group and Zhuanggu Guanjie Wan group compared with the model group (P < 0.01), in a time-dependent manner. Immunohistochemical staining indicated that the positive cels of urokinase-type plasminogen activator and urokinase-type plasminogen activator receptor were significantly decreased, but plasminogen activator inhibitor was significantly increased in theTougu Xiaotong capsule group andZhuanggu Guanjie Wangroup in a time-dependent manner. Western blot assay results had an identical trend to immunohistochemistry. These indicated thatTougu Xiaotong capsule showed preventive and therapeutic effects on osteoarthritis by regulating urokinase-type plasminogen activator system.
ABSTRACT
BACKGROUND:Tougu Xiaotong Capsule has pretty good clinical therapeutic effect on osteoarthritis of early and middle periods. However, the mechanism of Tougu Xiaotong Capsule is not ful y clarified. The RhoA GTPases can regulate chondrocyte apoptosis and hypertrophy. OBJECTIVE:To observe the Tougu Xiaotong Capsule on the expression of Rac1and Cdc42 in tumor necrosis factor-α-induced in vitro cultured rat articular chondrocytes, and to explore its mechanism of action for combating osteoarthritis. METHODS:Knee cartilage of the 4-week-old Sprague-Dawley rats was used to stably establish in vitro culture system of chondrocytes. Passage 3 chondrocytes were identified by toluidine blue staining. Chondrocyte apoptosis was successful y induced by 20μg/L tumor necrosis factor-αand then Tougu Xiaotong Capsule at different dosage (500, 100, 20 mg/L) was given after 24-hour incubation. MTT assay was used to detect cellsurvival, flow cytrometry to measure mitochondrial membrane potential, and western blot assay to determine the protein expression of Rac1, Cdc42, Bax and Bcl-2. RESULTS AND CONCLUSION:Tougu Xiaotong Capsule could reduce tumor necrosis factor-α-induced apoptosis of chondrocytes to improve the survival rate of the cells, and at the same time, could down-regulate the protein expression of Rac1, Cdc42 and Bax and increase the protein expression of Bcl-2 significantly (P<0.05). Tougu Xiaotong Capsule possibly plays a therapeutic efficacy on osteoarthritis by reducing promote apoptosis Rac1, Cdc42 and Bax expression and increasing apoptosis inhibiting gene Bcl-2 expression, thereby to inhibit apoptosis of chondrocytes.
ABSTRACT
<p><b>OBJECTIVE</b>To develop a HPLC method for determining the content of protopine in Corydalis racemose.</p><p><b>METHOD</b>Analysis was performed on a Gemini C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetonitrile-water containing 0.8% triethylamine and 3% acetic acid acetum (20:80) as the mobile phase. The flow rate was 1.0 mL x min(-1). The detection wavelength was 289 nm.</p><p><b>RESULT</b>The average content of protopine in Herb of Racemose Corydalis was 0.905%. The calibration curve of protopine was linear between 0.124-1.36 microg (r = 0.9999). The average recovery was 98.49% with RSD 1.9%.</p><p><b>CONCLUSION</b>This method is simple, reproducible and can be used to determine the content of protopine in C. racemose.</p>