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Objective To investigate the solubility and stability of Tetrodotoxin (TTX) in different solvents, and the effect of temperature and pH on its stability. Methods Solutions of TTX in different matrices were prepared. Their concentrations at different temperatures and pH buffers were determined by high performance liquid chromatography and their solubility and stability were analyzed and calculated. Results TTX was most soluble at pH 3.5 and its solubility decreased as the pH increased. TTX degraded most rapidly under strong alkali conditions, with complete degradation after 20 min of reaction at 0.1 mol/L sodium hydroxide and 70 ℃. The stability test results similarly demonstrated that TTX was least stable under alkaline conditions. In a PBS buffered solution at 37 ℃, pH 7.4, TTX concentration began to decrease consistently at 1~10h, with a degradation rate of 88.07±0.27% after 28 days. Conclusion TTX is readily soluble in acidic aqueous solutions at pH 3.5 and almost insoluble in alkaline aqueous solutions. Its stability is closely related to the temperature and pH of the medium. It is more stable in acidic aqueous solutions and easily degrades under alkaline conditions, and its degradation process could be accelerated by increasing temperature.
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Objective To explore the effects of Cordyceps sinensis extract (CSE) on osteoporosis and RANKL-mediated osteoclastogenesis. Methods Bone marrow-derived macrophages (BMMs) was isolated from the bone marrow of C57BL/6 mice. CSE was added in osteoclast differentiation. Osteoclasts were stained by tartrate-resistant acid phosphatase (TRAP). The nearly mature osteoclasts were planted on hydroxyapatite plates and the area of bone lacunae was observed by microscope. The F-actin belt was stained by DAPI and phylloeptide and the number of nuclei was observed by confocal microscopy. The expressions of DC-STAMP, ATP6V0D2, TRAP, CTSK, and NFATC1 were detected by q-PCR. The protein expression of the MAPK pathway was detected by Western Blot. The in vivo experiments were carried out by administering CSE to the ovariectomized mice daily through gavage. After 6 weeks of intervention, mouse femurs were taken for morphological analysis. Peripheral blood was taken for ELISA. Results CSE represses osteoclastogenesis, bone resorption, F-actin belts formation, osteoclast specific gene expressions and MAPK signaling pathways in vitro. In vivo study indicated that CSE prevents OVX-induced osteoporosis and preserves bone volume by repressing osteoclast activity and function. It also increases the serum ALP, BGP content, and reduces TRAP content. Conclusion CSE can attenuate osteoclast formation and OVX-induced osteoporosis, suggesting potential clinical therapeutic effects for osteoporosis.
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Aim To investigate the protective effect of riboflavin on ischemia brain damage and the mecha-nism.Methods The in vivo experiments were pro-cessed in male SD rats .Rats were randomly arranged into control group , model group and riboflavin group . The rats in riboflavin group were intraperitoneally in-jected riboflavin at the dose of 1 mg? kg -1 for seven consecutive days .Then the rats in model and riboflavin groups were carried out middle cerebral artery occlu-sion( MCAO) operation.After 24 h, all rats were sacri-ficed and the brain tissue was dissected to observe the infarct area, the edema and the ultrastructure damage . The brain tissue was dyed by triphenyl tetrazolium chloride .The brain edema was observed by the weight of ischemia-side semi-brain.The ultrastructure was ob-served by electron microscope .The in vitro experiments were processed in primary culture neurons by exposed to oxygen and glycose deprivation ( OGD) .The viability of neurons was assayed by MTT method .The enzyme activity of superoxide dismutase ( SOD) , catalase ( CAT)and glutathione peroxidase ( GSH-Px ) was assayed to explore the mechanism .Results Riboflavin signifi-cantly decreased the infarct area ( P<0.01 ) , inhibited the brain edema ( P <0.01 ) and inhibited the ultra-structure damage in rats after MCAO;riboflavin protec-ted the viability ( P <0.01 ) and the ultrastructure of neurons exposed to OGD .The enzyme activity of an-tioxidant SOD1 ( P <0.01 ) , CAT ( P <0.01 ) and GSH-Px ( P <0.01 ) was protected by riboflavin in MCAO model .No difference was found in the activity of SOD2 . Conclusion Riboflavin inhibits ischemia brain damage , and the protection of the activity of an-tioxidants is involved in the mechanism .
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ObjectiveTo investigated the effects of Rhodiola Compound on improving the intellective function in mice and provide the basis for clinical application.MethodsMice were divided to different groups of three doses of rhodiola compound (0.3 g/kg,0.6 g/kg,1.2 g/kg) and swimming abilities were tested.Other mice were administrated single dose of compound rhodiola( 1.2 g/kg) and training by Morris water maze.Drug's improving intelligence function was assessed using memory acquisition impaired models made by scopolamine or alcohol.When the Morris water maze test was finished,mice were killed and brains were removed immediately to measure SOD and NO levels.ResultsGroups of three doses of compound rhodiola could significantly prolong the swimming time(P ＜ 0.05,P ＜ 0.01 ).Compound Rhodiola group can significantly reduce the swimming distance than the untreated group( ethanol model group:(26 906.6 ± 2769.7 ) mm,RCE treated group:( 19 586.1 ± 6826.7 ) mm ; P ＜0.05 ).Swimming distance and time of cross-platform quadrant was significantly increased,comparing with model group (P ＜ 0.05 ).Compound Rhodiola significantly enhanced the activity of mouse brain's SOD ( Scopolamine model group:( 150.3 ± 17.7 ) U/ml,RCE treated group:( 197.9 ± 16.8 ) U/ml ; P ＜ 0.05 ) and NO levels ( Scopolamine model group:( 44.7 ± 16.7 ) μmol/gprot,RCE treated group:( 65.4 ± 14.5 ) μmol/gprot ; P ＜ 0.05 ) significantly.ConclusionCompound Rhodiola could promote mice learning and memory function,SOD and NO in brain maybe play a important role in this effect.
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Objective To study the antitumor activities of extract from Ligia exotica(Roux).Methods The dried powder of total Ligia exotica(Roux) was extracted by 37 ℃ water.The solution was concentrated in vacuum,and then was freeze-dried to afford crude extract.The inhibitory effect of the extract on tumor cells proliferation was assayed by MTT method,and transplant tumor model of sarcoma 180(S180) was used.Results The extract from Ligia exotica displayed obvious proliferation inhibitory effect on HeLa,7901,NCI cells,and no growth inhibitory effect on 929 cells in vitro.After administration at the doses of 0.25,0.50,1.00 g?kg-1,ip,for 7d in tumor-bearing mice with S180,The extract caused 26.9 %,45.3 %,64.6 % inhibition rates,respectively.Conclusion The extract from Ligia exotica showed significant antitumor activity.
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AIM:To estimate the toxicological characterization of Jiutong Capsule(Radix Puerariae lobatae,Fructus seu Semen Hoveniae,etc.).METHODS:The acute toxicity test,micronucleus test of bone marrow cell,sperm shape abnormality test in mice,the Ames test,and 30 days feeding test in rat were performed.RESULTS:(1) The acute toxicity test showed that LD_ 50 of Jiutong Capsule was more than 10.0 g/kg.(2) The result of Ames test,micronucleus test of bone marrow cell,and sperm shape abnormality test were negative.(3) The 30 days feeding test showed that Jiutong Capsule had no cumulate toxicity in mice.CONCLUSION:The results show that Jiutong Capsule does not have toxicity,and it can't cause mutation and heredity toxicity.