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OBJECTIVE To investigate the effect and mechanisms of chrysophanol on the expression of c-fos and c-jun mRNA induced by ammonia chloride(NH4Cl) in mouse astrocytes. METHODS Primary mouse astrocytes were cultured with NH4Cl 5 mmol·L-1+chrysophanol 0.1,1.0 or 10.0 mg·L-1 ,or NH4Cl 5 mmol · L- 1+extracellular regulated kinase1/2(ERK1/2) inhibitor UO126 10 mmol · L- 1 and NH4Cl 5 mmol · L-1+p38 mitogen-activated protein kinase(p38 MAPK)inhibitor SB239063 10 mmol · L-1 for 48 h. The levels of glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),malondaldehyde(MDA), nitric oxide(NO)and nitric oxide synthase(NOS)were detected by UV spectrophotometry. The phos?phorylation levels of ERK1/2 and p38 MAPK were detected by Western blotting. The expression of c-fos and c-jun mRNA was detected by RT-PCR. RESULTS Compared with NH4Cl 5 mmol · L- 1 group, ammonia-induced astrocytes oxidative stress was improved by chrysophanol (1.0 and 10.0 mg · L-1). The content of MDA and NO and the activity of NOS were reduced(P<0.05). The activity GSH-Px and SOD was increased(P<0.05). The phosphorylation level of ERK1/2 and p38 MAPK induced by ammonia was reduced in chrysophanol groups (P<0.05). Ammonia-induced c-fos and c-jun mRNA expression downregulation was reversed by chrysophanol (0.1,1.0 and 10.0 mg · L-1)and UO126 and SB239063(P<0.05). CONCLUSION Chrysophanol may improve the downregulated expresion of c-fos and c-jun mRNA in mouse astrocytes exposed to NH4Cl by anti-oxidative stress by inhibiting the ERK1/2 and p38 MAPK phosphorylation.
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Aim To investigate the protective effects of chrysophanol( Chry) on immune function of lead poi-soning mice. Methods The lead poisoning model was established by peritoneally injecting mice with 7 mg · kg-1 lead acetate every day for 8 days. After Chry was ip injected for 14 days,spleen and thymus index, the white blood cell count, T, B lymphocyte proliferation, phagocytic function of macrophages, natural killer cell ( NK cell) activity were detected. The concentrations of IFN-γ,IL-2,IL-4,IL-10 in the lead poisoning mice ser-um were determined by enzyme-linked immunosorbent assay ( ELISA) . Results Intraperitoneal injection of 7 mg · kg-1 lead acetate for 8 consecutive days could cause an immunity decline in lead poisoning mice, Chry could significantly improve the immunity of lead poisoning mice. Compared with model group, Chry sig-nificantly improved growth rate, viscera index and white blood cell count of lead poisoning mice at differ-ent extent ( P0. 05 ) , only IL-10 was significantly increased in Chry ( 0 . 1 mg · kg-1 ) treatment group ( P<0 . 05 ) . Conclusion Chry can significantly improve the im-mune function of lead poisoning mice.
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AIM:To establish a method for determining saikosaponin A,paeoniflorin,hesperidin and ferulic acid in Chaihu Shugan Powder(Pericarpium Citri Reticulatae,Radix Bupleuri,Rhizoma Chuanxiong,Fructus Aurantii,Radix Paeoniae alba,etc.).METHODS:The chromatographic saparation was performed on a Hypersil C_(18) column(250 mm×4.6 mm,5 μm).The mobile phase was acetonitril-water(43:57)for saikosaponin A,and the mobile phase for rest components was acetonitrile-0.5% acetic acid(40:60).All of flow rates were 0.8 mL/min and column temperature maintained at 30℃.The detection wavelength was set at 200-400 nm.RESULTS:The four constituents were separated within 15 min.The linear ranges of saikosaponin A,paeoniflorin,hesperidin and ferulic acid were 38.5-166.7 μg/mL(r = 0.999 6),15.9～254.5 μg/mL(r = 0.999 9),22.1-353 μg/mL(r =0.999 3),6.30-201.5 μg/mL(r =0.999 9),respectively.The average recoveries were 97.57%,97.40%,98.86%,96.37%,respectively.The RSD were 2.1%,1.1%,0.70%,1.3%,respectively.CONCLUSION:The method is rapid,simple and accurate,and can be used for quality control of Chaihu Shugan Powder.
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Objective To optimize the preparation technology of chrysophanol loaded polybutylcyanoacrylate nanocapsules by interfacial polymerization,sieve the optimal technology conditions of preparation,and study its quality. Methods Based on the single factor experiment,the formulation was optimized by L9(34) orthogonal design with entrapment efficiency as reference index. The factors including agitating speed,pH value of aqueous phase,amount of ?-butylcyanoacrylate (BCA) and of ethyl acetate were screened. At last,its shape,particle size,the encapsulation efficiency,and the drug loading were investigated as its quality inspection. Results When the ammunt of chrysophanol was 5 mg,the optimum preparation conditions were established as follows:the agitating speed was 800 r/min,the pH value of aqueous phase was 2,the amount of ?-BCA was 13 ?L and ethyl acetate was 0.6 mL. Under this optimum condition,the mean entrapment ratio was 82.19%,and the mean drug loading was 21.48%,the mean diameter of the nanocapsule was 246 nm. The particle size was well-distributed showed by electron microscope pictures. Conclusion The prepared chrysophanol loaded polybutylcy-anoacrylate nanocapsules has small size,high encapsulation efficiency,and drug loading. The preparation technique established in this study is stable and feasible. It also can be used for iv injection.
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AIM:To establish a method for determining saikosaponin A,paeoniflorin,hesperidin and ferulic acid in Chaihu Shugan Powder(Pericarpium Citri Reticulatae,Radix Bupleuri,Rhizoma Chuanxiong,Fructus Aurantii,Radix Paeoniae alba,etc.).METHODS:The chromatographic saparation was performed on a Hypersil C18 column(250 mm?4.6 mm,5 ?m).The mobile phase was acetonitril-water(43:57) for saikosaponin A,and the mobile phase for rest components was acetonitrile -0.5% acetic acid(40:60).All of flow rates were 0.8 mL/min and column temperature maintained at 30 ℃.The detection wavelength was set at 200-400 nm.RESULTS:The four constituents were separated within 15 min.The linear ranges of saikosaponin A,paeoniflorin,hesperidin and ferulic acid were 38.5-166.7 ?g/mL(r=0.999 6),15.9～254.5 ?g/mL(r=0.999 9),22.1-353 ?g/mL(r =0.999 3),6.30-201.5 ?g/mL(r=0.999 9),respectively.The average recoveries were 97.57%,97.40%,98.86%,96.37%,respectively.The RSD were 2.1%,1.1%,0.70%,1.3%,respectively.CONCLUSION:The method is rapid,simple and accurate,and can be used for quality control of Chaihu Shugan Powder.
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OBJECTIVE:To study the best preparation technology of Gross Saponins tribullu terrestris droping pill. METHODS: The ratio of Gross Spongines Tribullu terrestris to matrix, dripping temperature, dripping distance and dripping speed were investigated by preliminary test and orthogonal test with sphericity, the variation coefficient of weight of pill,time limit of dissolution and appearance of the dropping pill were taken as scoring indexes. Meanwhile, the technical conditions were optimized by orthogonal test on the basis of preliminary test.RESULTS: The optimal technical conditions were as follows: the ratio of matrix to Gross Saponins Tribullu terrestris was 5∶1;the dripping temperature was 85 ℃;the dripping speed was 8 d?min-1 and the dripping distance was 12 cm.CONCLUSION:The technology is simple, feasible and the evaluation indexes were reliable and reasonable and it can meet the requirement for dripping pill specified in China Pharmacopeia (2005 Edition).
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Objectives To examine sICAM-1 in Pneumocystis carinii pneumonia(PCP) and the effect of TMP-SMZ therapy on its level and on pathological and immunological changes in rats.Methods 50 female Wistar rats were randomly divided into normal group(N group),PCP model group(PCP group) and TMP-SMZ therapy group(SMZ group).1 mg of dexamethasone was injected intramuscularly twice a week for rats in PCP and SMZ groups to induce PCP.Normal saline was injected for N group in the same way.When the infection was confirmed,TMP-SMZ was given to rats in SMZ group by 25 mg/(kg.d) for 5 days for 3 courses with an interval of 2 weeks.sICAM-1 in serum was detected by ELISA,and the pathological changes in lungs and liver and the Pc in alveoli of lungs were observed.Results The level of sICAM-1 in PCP and SMZ groups at the 3rd week [(1.847?0.50) ng/ml,(1.787?0.59) ng/ml] was lower notably than that at 0 week[(2.407?0.81) ng/ml,[(2.478?0.59) ng/ml respectively](P0.05).Conclusion The sICAM-1 level in rats was low but significantly increases after the induction of PCP.