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Objective To study effect of Tongbiling (TBL) on VEGF mRNA expression in RA synovlcytes cultured in vitro and explore its mechanism. Method Synovial cells from the knee joint in patient with RA were digested, divided and separately cultured after arthroscopy. Blank controls:DMEM culture containing 10% FBS in saline;Low-dose admission group of TBL:DMEM culture containing 10% FBS in 50 mg/L TBL;High-dose admission of TBL:DMEM culture containing 10% FBS in 200 mg/L TBL. RNA were distilled. Expression of VEGF mRNA were detected with RT-PCR. Result Compared with control group, expression of VEGF mRNA in low and high-dose admission groups of TBL both decreased (P
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Objective: The study was to reveal the inhibition of neovascularization by Tongbiling. Methods: Rat model were established by injecting CFA at the root of the tail and divided into model control group, Indomethacin group and Tongbiling group at random. The serum and paws were collected after seven days treatment. 10ng/L VEGF solution was added I into CAM to stimulate the neovascularization, and to observe the influence of each group of serum on the first and second order new vessels. RT-PCR method was taken to determine the expression of mRNA of VEGF and VEGFR-1 of paws in different groups. Results: The number of vessels in the model control group was significantly more than that in the Indomethacin group and the Tongbiling group (P
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[ Objective ] To establish a method for the determination of serum strychnine concentration in rheumatoid arthritis patients treated with Tongbiling prescription and to monitor the serum strychnine concentration. [ Methods ] Serum strychnine was extracted with solid phase extraction column. Reversed-phase HPLC conditions were: Hypersil Division ODS C18 column (150 mm ?4.6 mm, 5?m), mobile phrase being methanol: water (70 : 30) with 0.6 g potassium dihydrogen phosphate and 1.0 g dodecyl sodium sulfonate added in 1000 mL mixture (adjust pH to 4.5 with acetic acid), detecting wavelength at 254nm and a flow rate of 1.0 mL/min at room temperature. [Results] Strychnine has a good linearity in the range of 0.031- 2.000?g?mL-1. The minimum detecting concentration was 0.01?g?mL-1 and the average recovery was over 90%. Intra-day RSD and inter-day RSD ranged from 1.53% to 5.32%. [Conclusion] This method is simple and efficient, and can be used for the determination of strychnine.
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[ObjectivejTo explore the therapeutic mechanism of Tongbiling (TBL) prescription for rheumatoid arthritis (RA) by observing the effects of TBL on inflammation and expression of vascular endothelial growth factor (VEGF) in synovium of rats with collagen-induced arthritis (CIA) .[Methods]CIA rat models were established by subcutaneous injection of type II collagen. Pathological sections of synovium in knee joint were made after medication for 37 days and then stained by HE method and immunohistochemical method respectively. Infiltration of inflammatory cells and hyperplasia of macrophages A and fibrous tissue in HE-stained pathological sections were observed under microscope and expression of VEGF was observed by semi-quantitative immunohistochemical method. [ Results ] Infiltration of inflammatory cells and hyperplasia of macrophages A and fibrous tissue in high-dose TBL group were inferior to those in model group (P 0.05); both TBL and Radix Tripterygii Wilfordii at high dose decreased the expression of VEGF (P
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[Objective] To observe the effects of Tongbiling (TBL) prescription in counteracting cartilage destruction in rats with rheumatoid arthritis (RA). [Methods] Collagen-induced arthritis (CIA) rat models were established to observe the therapeutic effects of TBL. After a short-term treatment, expressions of tumor necrosis factor (TNF - a) and transforming growth factor (TGF- P) in cartilage tissue of model group, methotrenxate (MTX) tablets group and Radix Tripterygii Wilfordii (RTW) preparation group were examined by immunohistochemical semi-quantitative assessment method. [ Results ] The number of chondrocytes with positive TNF - a expression was lower and the chondrocytes were stained lighter in normal group than those in model group and low-dose MTX group ( P
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[Objective] To establish a method for the determination of brucine and strychnine contents in Tongbiling Tablet. [Methods] Reversed-phase HPLC conditions were: Nucleodur C18 Gravity column (125 mm ? 4.6 mm, 5?m), mobile phrase being methanol-water (60:40) with 0.5 g potassium dihydrogen phosphate and 1.0 g dodecyl sodium sulfouate and 0.5 mL triethylamine in 1 000 mL mixture (adjusting pH to 4.5 with acetic acid) , detecting wavelength at 254 run and a flow rate of 1.0 mL/min at room temperature. [ Results ] The average recovery of brucine was 97.69 % ( RSD = 2.03%) and that of strychnine 97.93% ( RSD = 2.61%). [Conclusion] This method is efficient and can be used for the quality control of Tongbiling Tablet.