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AIM: To investigate the effects of Rhodiola sachalinensis on ocular blood flow in diabetic retinopathy rats. METHODS:A total of 90 SD rats were randomly divided into control group (n=30), model group (n= 30) and intervention group (n=30). Rats in the model group and intervention group were fed with high glucose and high fat diet and injected with streptozotocin (40mg/kg) in order to construct DR model rats,while rats in the control group were fed with basic diet and injected with the same amount of normal saline. After 4wk, the rats in the intervention group were injected with Rhodiola sachalinensis injection(10mL,one per day),while rats in the control group and the model group were injected with normal saline. The course of intervention treatment was 4wk. The ocular blood flow in rats was detected by laser doppler flowmetry, the apoptosis of retinal cells in rats was detected by TUNEL method, the expression of vascular endothelial growth factor (VEGF), glial fibrillary acidic protein ( GFAP ) and glutamate/aspartate transporter (GLAST) in retina of rats was detected by RT-PCR method and Western blot method. RESULTS:The whole blood viscosity,whole blood high shear viscosity,low shear blood viscosity and erythrocyte sedimentation of rats in the model group and intervention group were higher than those in the control group (P<0 01),while the indexes of intervention group were lower than that of model group (P<0.01). The PSV and EDC of rats in the model group and intervention group were lower than those in the control group (P<0.01), and the Rl was higher than that in control group (P<0.01). The PSV and EDC in the intervention group were higher than those in the model group (P<0.01), and Rl was lower than that in the model group (P<0.01). The apoptosis rate of retinal cells of model group and intervention group was higher than that of the control group (P<0.01), that in the intervention group was lower than that in the model group (P<0.01). The expression of VEGF and GFAP in retina tissue of model group and intervention group was higher than that of the control group (P<0.01), and the expression of GLAST was lower than that in the control group(P<0 01). The expression of VEGF and GFAP in the intervention group was lower than that in the model group(P<0.01),and the expression of GLAST was higher than that in the model group (P<0.01). CONCLUSION: Rhodiola sachalinensis injection can significantly improve the ocular blood flow in diabetic retinopathy rats, reduce the apoptosis of retinal cells in rats, down regulate the expression of VEGF and GFAP, and up regulate the expression of GLAST.
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Objective: To optimize the sulfated modification conditions of Rhodiola sachalinensis polysaccharide (RSP) and improve the anti-oxidant activity of RSP. Methods: RSP were sulfated by chlorosulfonic acid-pyridine method, and the optimal conditions for the sulfated modification were determined by single factor experiments. The physical and chemical properties of RSP and sulfated RSP (S-RSP) were analyzed by infrared spectroscopy (IR) and scanning electron microscopy (SEM). The relation between substituting degree (DS) of S-RSP and anti-oxidative activity of polysaccharide was investigated by testing the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging ability of RSP and S-RSP. Results: When the volume ratio of chlorosulfonic acid to pyridine was 1∶4, the reaction time was 2 h and the reaction temperature was 60 ℃, the maximum sulfur content of S-RSP was 18.83% and the DS was the highest for 2.38. Moreover, the anti-oxidant activity of RSP was enhanced by the sulfated modification, and there was a certain positive proportional relationship between DS and DPPH free radical scavenging ability of S-RSP. Conclusion: The volume ratio of chlorosulfonic acid to pyridine affects the DS of S-RSP, and the sulfated modification can increase the anti-oxidant capacity of RSP by changing its polarity.
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Objective: To investigate the chemical constituents from the roots and rhizomes of Rhodiola sachalinensis. Methods: The chemical constituents were isolated by repeated silica gel chromatography, medium pressure column chromatography, and semi-preparative liquid chromatography, and their structures were elucidated by chemical properties and spectroscopic analyses. Results: Eighteen compounds were isolated and identified to be gallic acid (1), p-hydroxybenzoic acid (2), salidroside (3), benzyl-O-β-D-glucopyranodide (4), phenylethyl-8-O-β-D-glucopyranodide (5), cinnamyl-β-D-glucopyranoside (6), sachalinol (7), quercetin (8), quercetin-3-O-β-D-glucopyranoside (9), kaempferol (10), kaemferol-7-O-α-L-rhamnopyranoside (11), kaempferol- 7-O-β-D-glucopyranoside (12), kaemnpferol-3-O-α-L-rhamnoside (13), kaempferol-3-O-β-D-glucopyranoside-7-O-α-L-rhamnoside (14), tricin (15), tamarixetin (16), herbacetin-7-O-α-L-rhamnoside (17), and herbacetin-3-O-β-D-glucopyranoside-7-O-α-L- rhamnoside (18). Conclusion: Compounds 9, 12, and 16 are obtained from the plants in Rhodiola L. for the first time. Compounds 2, 7, 8, 14, and 18 are obtained from this plant for the first time.
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Objective: To obtain autotetraploid clones of Rhodiola sachalinensis. Methods: Germplasm with doubled chromosomes was acquired according to colchicine soaking seeds, and ploidy was identified according to stomata size and density measurements, chromosome counting and flow cytometry analysis. Chimera was purified after several times of regenerations. Results: Colchicine concentration and treatment duration had significant effect on seed germination rate, seedling death rate and ploidy mutation rates, and they were 18.2%, 73.6%, and 68.7%, respectively after the seeds were soaked in 0.2% colchicine solution for 72 h. Stomata diameter was promoted and density was declined significantly in morphological variation plantlets; normal diploid chromosome number was 2n = 2x = 26, and tetraploid chromosome number was 2n = 4x = 52. Meanwhile, aneuploidy was found in the population of plantlets after colchicine soaking, leaves of ploidy mutation plantlets were used as expiants to regenerate to tube plants, and homozygous clones were obained after three regeneration and purification cycles. There was no chimera in the homozygous clones according to flow cytometry analysis. Conclusion: It is feasible to obtain autotetraploid according to colchicine treatment on seeds of R. sachalinensis.
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OBJECTIVE:To compare the contents of Rhodioloside and polysaccharide extracted from different part in cultivated Rhodiola sachalinensis.METHODS:The Rhodioloside was extracted with 70% alcohol and determined by HPLC;The polysaccharide was extracted with water and determined by sulphuric acid-phenol method.RESULTS:The content of the Rhodioside ranged between 0.026 5% and 0.443 2%,and the content of polysaccharide between 0.965% and 4.612% in the medicinal herbs,with the main root showing the highest contents.CONCLUSION:The contents of Rhodioside and polysaccharide are obviously different in different part of cultivated Rhodiola sachalinensis.The results provided reasonable theoretical evidence for the exploitation of cultivated Rhodiola sachalinensis.
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Peripheral insulin resistance in obese/type II diabetes animals results from an impairment of insulin-stimulated glucose uptake into skeletal muscle. Insulin stimulate the translocation of GLUT4 from intracellular location to the plasma membrane. Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) is implicated in mediation of fusion of GLUT4-containing vesicle with the plasma membrane. Present study investigated regulatory effects of Rhodiola sachalinensis administration and exercise training on the expression of GLUT4 protein and SNAREs protein in skeletal muscles of obese Zucker rats. Experimental animals were randomly assigned into one of five groups ; lean control (LN), obese control (OB), exercise-treated (EXE), Rhodiola sachalinensis-treated (Rho), combine of Rho & EXE (Rho-EXE). All animals of exercise training (EXE, Rho-EXE) performed treadmill running for 8 weeks, and animals of Rho groups (Rho, Rho-EXE) were dosed daily by gastric gavage during the same period. After experiment, blood were taken for analyses of glucose, insulin, and lipids levels. Mitochondrial oxidative enzyme (citrate synthase, CS ; beta-hydroxyacyl-CoA dehydrogenase, beta-HAD) activity were analysed. Skeletal muscles were dissected out for analyses of proteins (GLUT4, VAMP2, syntaxin4, SNAP23). Results are as follows. Exercise and/or Rhodiola sachalinensis administration significantly reduced body weight and improved blood lipids (TG, FFA), and increased insulin sensitivity. Endurance exercise significantly increased the activity of mitochondrial enzymes and the expression of GLUT4 protein, however, administration of Rhodiola sachalinensis did not affect them. The effect of exercise and/or Rhodiola sachalinensis administration on the expression of SNARE proteins was unclear. Our study suggested that improvement insulin sensitivity by exercise and/or Rhodiola sachalinensis administration in obese Zucker rats is independent of expression of SNARE proteins.
Subject(s)
Animals , Rats , Body Weight , Cell Membrane , Glucose Transporter Type 4 , Glucose , Insulin Resistance , Insulin , Muscle, Skeletal , Negotiating , Obesity , Oxidoreductases , Rats, Zucker , Rhodiola , Running , SNARE Proteins , Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins , Vesicle-Associated Membrane Protein 2ABSTRACT
Objective To isolate protoplasts from tube plant leaves of Rhodiola sachalinensis and regenerate plantlets after protoplast culture.Methods Preculture treatment and size of explants,enzyme concentration,mannitol concentration in enzyme mixture related with protoplasts isolation were studied to determine the superior optimized conditions.Results Explants could be used to isolate protoplast without dark preculture,and leaf length should be longer than 1.5 cm.The best incubating enzyme solution contains 1.0% cellulase Onzuka R-10,0.5% Macerozyme R-10,10 mmol/L CaCl2?2H2O,0.1% MES,0.7 mmol/L KH2PO4,and 0.5 mol/L mannitol.The enzyme and explants mixture were shaken for 4 h at 25 ℃.The protoplasts yield and viability were 39.43?106/g fresh weight and 78.6%,respectively.Purified protoplasts were cultured in medium 1/2 MS+1 mg/L 2,4-D+0.5 mg/L ZT+0.5 mol/L mannitol +500 mg/L hydrolysis of casein initially with shallow liquid layers,and calli formed within 40 d.After calli were transfered to MS+1 mg/L 6-BA+0.1 mg/L NAA,adventitious buds were induced from calli.Shoots longer than 2 cm rooted within 30 d when they were transfered to 1/2 MS medium.Conclusion The study provides the scientific base for protoplast fusion in polyploidy breeding of R.sachalinensis.