Protective effect of astragalosides IV on retinal pigmentepithelium injury induced by methylglyoxal / 中国药理学通报

Aim To investigate the protective effect of astragaloside IV (AS-Ⅳ) on human retinal pigment epithelium injury induced by methylglyoxal (MGO), and explore its molecular mechanism.Methods The injury of ARPE-19 cells was induced by MGO and the cell viability was measured by CCK-8 method.The morphology of cell nucleus was analyzed by Hoechst 33342 staining and the cell apoptosis was analyzed by flow cytometry to detect labbled Annexin V-FITC/PI.JC-1 staining and fluorescence probe DCFH-DA were employed to evaluate the change of mitochondrial membrane potential and reactive oxygen species (ROS).The levels of SOD, MDA, caspase-9 and caspase-3 were determined by respective kits.Western blot was used to analyse the expression of Bcl-2, Bax and PARP.Results AS-Ⅳ could significantly inhibit the decrease of cell viability induced by MGO, improve the morphology of cell nucleus, reduce the ARPE-19 cell apoptosis rate and the level of ROS and MDA, and increase the activity of SOD.Furthermore, AS-Ⅳ could enhance mitochondrial membrane potential, the ratio of Bcl-2/Bax and the expression of PARP, and inhibit the activation of caspase-9 and caspase-3.Conclusion AS-Ⅳ may protect ARPE-19 cells from the injury induced by MGO by increasing the antioxidant ability of ARPE-19 cells and inhibiting cell apoptosis.

Inhibitory Effects ofQi-Zhu Granules on Early Proteinuria in Diabetic Nephropathy Rats with Syndrome ofQi-yin Deficiency and Phlegm Blocking Collaterals / 世界科学技术-中医药现代化

ThisstudywasaimedtoobservetheinhibitoryeffectsofQi-Zhu (QZ)granulesonearlyproteinuria in diabetic nephropathy rats with syndrome of qi-yin deficiency and phlegm blocking collaterals. A total of 44 rats were randomly divided into the blank group, model group, Huang-Kui capsule group, QZ granules group. The rat model of diabetic nephropathy with syndrome of qi-yin deficiency and phlegm blocking collaterals was induced by the combination of unilateral renal artery ligation, diet of high-calorie and high cholesterol, and intraperitoneal injection of low-dose streptozotocin. The medication was given for 8 weeks. The concentrations of protein and creatinine in urine were observed on the 4th week. The blood glucose, blood lipids, liver function and renal pathological changes were observed at the end of the experiment. The results showed that compared with the model group, QZ granules can obvious suppress early proteinuria in diabetic nephropathy, promote creatinine excretion, regulate blood lipid metabolism, protect liver function and improve renal pathological changes. It was concluded that QZ granules had independent inhibition effect on early proteinuria in diabetic nephropathy rats with syndrome of qi-yin deficiency and phlegm blocking collaterals. The effect was independent of lowing blood glucose. It represented the corresponding relation between the syndrome and efficacy in Chinese herb compounds.

Content Determination of Berberine Hydrochloride in Jiangtang Xiaozhi Tablets by HPLC / 中国药房

OBJECTIVE:To establish a method for the content determination of berberine hydrochloride in the Jiangtang xiao-zhi tablets. METHODS:HPLC was conducted with the Symmetry C18 column. The mobile phase was acetonitrile-0.05 mol/L sodium dihydrogen phosphate(pH adjusted to 3.0 using phosphoric acid)(24∶76,V/V)at the flow rate of 1.0 ml/min. The detection wave-length was 345 nm,the column temperature was room temperature and injection volume was 10 μl. RESULTS:The linear range of berberine hydrochloride was 0.522-4.698 μg(r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 0.72%;the average recovery was 97.79%(RSD=2.09%,n=6). CONCLUSIONS:The method is simple,accurate,sensitive and reproducible,and can be used for the content determination of berberine hydrochloride.

Trypsin digestion method for the long-term formalin-fixed retinal in rats / 中国药理学通报

Aim To investigate the optimum trypsin digestion time and the optimum trypsin concentration of the long-term formalin-fixed retina in rats.Methods Male SD rats were sacrificed by cervical dislocation. The removed eyes were fixed in 4% formaldehyde fixa-tive for 2 days,2 weeks,4 weeks,3 months,5 months and 7 months respectively.Fixed retinas at dif-ferent time were separated and put in 3%,6% and 9% trypsin at 37℃ in incubator.The time required was recorded for complete digestion of retinas,which fixed for different time and digested in different con-centrations of trypsin.The retinal blood vessels were recorded under a microscope after PAS staining of stretched retinal blood vessel.Results After conven-tionally fixed for 48 hours,retinas were digested com-pletely in 3% trypsin at 37℃ for 3.5 hours.The di-gestion time extended to 9 hours and 1 4 hours for the 2 weeks and 1 month fixed eyes,using 3% trypsin, which achieved the same result as conventional fixed eyes.For the 3 months fixed eyes,a complete retinal blood vessel could also be attained by using conven-tional concentrations of trypsin,and only by extending digestion time to 22 hours.It is also indicated that the increased concentration of trypsin did not shorten the time of digestion.In addition,for the fixed eye from 5 months to 7 months,6% and 9% trypsin could short-en,to some extent,the time to 22 hours and 28 hours respectively,which was conductive to obtaining rela-tively complete retinal blood vessels.Conclusions Retinal tissues which fixed time shorter than 3 months can completely be digested using 3% trypsin.Howev-er,for retinal tissue,which fixed from 5 months to 7 months,both the extending time of digestion and the increasing concentration of trypsin to 6% are necessary to get optimum retinal vascular digestion.

Large-sample Analysis of the Normal Reference Value of Hematological and Serum Biochemical Parameters in Healthy Beagle Dogs / 中药新药与临床药理

Objective To establish the normal reference value range of hematological and serum biochemical parameters in Beagle dogs for our own research center and to supply references for establishing the national standard.Methods CV Beagle dogs aged 4～6 months were selected as the investigative objects.Of the Beagle dogs,263 were male with body weight of(8.09?0.78)kg,263 were female with body weight of(6.84?0.87)kg.Blood was collected from cephalic vein at the endgame of the experiment,then both hematological and serum biochemical parameters were measured.The acquired data were analyzed by the "Independent-Samples T Test" process of SPSS V 13.0.Regarded the males and the females as two independent samples,the difference between sexes was also compared.Results The mean normal reference values and the standard deviation of hematological and serum biochemical parameters in both male and female Beagle dogs were obtained by analysis.Conclusions Comparison of sexes showed that there exists obviously difference between male and female Beagle dogs referring to 3 serum biochemical parameters of T-Bil,GLU and ALB.