ABSTRACT
Objective: To observe the effect of phlegm and blood stasis on the expressions of sirtuin 3(SIRT3)protein and urate transporter 1(URAT1) mRNA in skeletal muscle of diabetic rats with gout. Method: The 40 healthy rats, excepting the normal group, the remaining groups were fed with high-fat diet combined with low-dose streptozotocin solution (40 mg·kg-1) once a day, with blood glucose "16.7 mmol·L-1" as the criterion for the diabetes model. After 4 days, the 5% sodium urate solution was injected into the joint cavity once to induce the gout model. After the successful modeling, the Biling group (10 g·kg-1), the indomethacin group (5 mg·kg-1) and the pioglitazone group (10 mg·kg-1) continued to be administered for 21 days. The normal group and the model group were given the same amount of normal saline. The expression of SIRT3 protein in skeletal muscle tissue was determined by Western blot, URAT1 mRNA expression in bone tissue was detected by quantitative real-time fluorescence polymerase chain reaction(Real-time PCR),and blood was collected to measure blood glucose (GLU), blood uric acid (UA) and C-reactive protein (CRP). Result: Compared with the normal group, GLU, UA and CRP in the model group were significantly increased (PPPPPPPPConclusion: Biling Qutong prescription with effects in purging turbidity, detoxifying and dredging collaterals can significantly reduce the content of serum inflammatory factor CRP, significantly increase the protein expression of SIRT3 in skeletal muscle tissue of model rats, lower the content of URAT1 mRNA, reduce the blood glucose and blood uric acid levels in diabetic gout rats, and protect joints.
ABSTRACT
<p><b>OBJECTIVE</b>To establish an HPLC method for analysis of bis(p-fluorobenzyl) trisulfide(BFTS) and bis(p-fluorobenzyl)disulfide(BFDS) in the lungs of rat.</p><p><b>METHODS</b>5.0 ml extract solvent (n-hexane: isopropyl alcohol=95:5, v/v) and 20 microl of 11.50 microg/ml dibenzyl disulfide (internal standard) were added to 0.2 g lung sample followed by homogenization. After centrifugation, 4.0 ml of supernatant was separated and vaporized to dryness, and the residue was reconstituted in mobile phase for HPLC analysis. The HPLC analysis was performed on an SB C18 column using acetonitrile and water (65:35, v/v) as mobile phase with a flow rate of 1.0 ml/min with UV detection at 220 nm.</p><p><b>RESULT</b>The calibration curves for BFTS and BFDS in sample were linear over the concentration ranges of 0.04712-14.78 microg/g(r=0.999) and 0.04831-23.96 microg/g(r=0.999), respectively. The limits of quantification were 0.04712 microg/g and 0.04831 microg/g for BFTS and BFDS, respectively. The assay recoveries for BFTS and BFDS ranged from 95.71%-107.2% and 90.00%-110.5%, respectively. The precisions were obtained with RSD of <10%. The developed method was successfully applied to study the content of BFTS and BFDS in the lungs of rats after intravenous injection of 12.5 mg/kg BFTS.</p><p><b>CONCLUSION</b>The method developed is simple, selective, repeatable and accurate, which can be applied to study the tissue distribution of BFTS and BFDS.</p>