Studies On The Anti-inflammatory Constituents Of Juniperus Sabina L. And Therapeutic Of The Effects Of Its Total Favonoids On Adjuvant Arthritis In Rats

Objective:To study the chemical constituents of Juniperus sabina L twigs, and the anti-inflammatory activity of its’compounds were screened, to set up the extraction process and purification process of its total flavonoids (TFJS), and to observe the preventive effect and therapeutic action of TFJS on Ajuvant Athritis (AA) in rat. Methods:Isolated and purified the chemical constituents by different kinds of column chromatography, and their structures were determined on the basis of the physicochemical properties and spectral analysis. To optimize the condition for extracting and the method for purifying TFJS. In order to evaluate the effects of TFJS of the AA in rats in the primary stage and the secondary stage, experimental animals of each experiment was divided into six groups respectively, set the control, the model, the positive(Tripterygium glycosides tablets) and the different doses of TFJS (125,250,500mg/kg) to cure the AA in rats. AA rat models were established by freund’s complete adjuvant. Some variances of rats were measured at different time, such as the weight change, the swelling degree in hind paws and the Arthritic Index (AI). The levels of interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-a (TNF-a) and ProstaglandinE2(PGE2) in serum of rats were examined by ELISA. Results:12compounds were obtained from Juniperus sabina L. The optimum extraction condition of TFJS was confirmed as follows:60%ethanol concentration, extracting time of26min, and liquid-material ratio of25:1(mL/g), the extraction rate of TFJS was53.25mg/g under this condition. The purification condition of TFJS was confirmed as follows:The sample density of1.645mg/mL, the sample volume of150mL with the adsorption velocity of1.5mL/min, the edulcoration volume of12BV, the eluent of70%ethanol of5BV with the elution velocity of1.5mL/min, the content of TFJS was69.54±1.00%with extraction rate of4.1%. The effects of TFJS on AA rats in three experiment, the swelling degree of inflammatory side hind paws in model group was significantly higher than the corresponding control group (P<0.05) at different time phases of experiment. With the does of TFJS increasing, the levels of IL-1β、IL-6TNF-α and PGE2in serum of the TFJS were lower than AA model group. In the secondary stage which the secondary swelling had been formed, the AI of model group was significantly higher than control group (P<0.01). With the does of TFJS increasing, AI were significantly lower than model group (P<0.05). Conclusions:12compounds were obtained from twigs and leaves of Juniperus Sabina L., of which isoscutellarein7-O-β-D-xylopyranose-(1→3)-α-L-rhamnoside (1) and hypolaetin1-O-β-D-xylopyranose-(1→3)-a-L-rhamnoside (2) was new constituent, isoquercetin (5) and hypolaetin7-O-β-D-glycoside(6) were firstly isolated from this genus. The extraction process and purification process of TFJS were stable and feasible, it can be helpful to the factory scale production. TFJS could inhibit and improve the symptoms of the primary and the secondary arthritis in AA rats.