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1.
Journal of Pharmaceutical Analysis ; (6): 88-98, 2023.
Article in Chinese | WPRIM | ID: wpr-991127

ABSTRACT

Aconitine,a common and main toxic component of Aconitum,is toxic to the central nervous system.However,the mechanism of aconitine neurotoxicity is not yet clear.In this work,we had the hypothesis that excitatory amino acids can trigger excitotoxicity as a pointcut to explore the mechanism of neurotoxicity induced by aconitine.HT22 cells were simulated by aconitine and the changes of target cell metabolites were real-time online investigated based on a microfluidic chip-mass spectrometry system.Meanwhile,to confirm the metabolic mechanism of aconitine toxicity on HT22 cells,the levels of lactate dehydrogenase,intracellular Ca2+,reactive oxygen species,glutathione and superoxide dismutase,and ratio of Bax/Bcl-2 protein were detected by molecular biotechnology.Integration of the detected results revealed that neurotoxicity induced by aconitine was associated with the process of excitotoxicity caused by glutamic acid and aspartic acid,which was followed by the accumulation of lactic acid and reduction of glucose.The surge of extracellular glutamic acid could further lead to a series of cascade reactions including intracellular Ca2+overload and oxidative stress,and eventually result in cell apoptosis.In general,we illustrated a new mechanism of aconitine neurotoxicity and presented a novel analysis strategy that real-time online monitoring of cell metabolites can provide a new approach to mechanism analysis.

2.
China Pharmacy ; (12): 1303-1308, 2020.
Article in Chinese | WPRIM | ID: wpr-821793

ABSTRACT

OBJECTIVE:To study the protective effects of Lespedeza cunea ta extract on glutamate-induced hippocampal cells HT22 injury of mice and its possible mechanism based on Nrf 2/HO-1 signaling pathway. METHODS :Using glutamate (5 mmol/L) to extablish the injury model of HT 22 cells. Using water soluble vitamin E as positive control (50 µmol/L),MTT assay was used to detect the effects of 0(blank control ),25,50,100 µg/mL petroleum ether extract ,dichloromethane extract ,ethyl acetate extract of L. cuneata on the proliferation of glutamate-induced injury cellsafter pretreated for 12 h. Using water soluble vitamin E as positive control (50 µmol/L),DCFH-DA assay was used to detect the effects of 0(blank control ),25,50,100 µg/mL L. cuneata dichloromethane extract on the level of active oxygen (ROS)in glutamate-induced injury cells after pretreated with 12 h. Using HO-1 agonist CoPP as positive control ,Western blotting method was used to detect the effects of 0(blank control ),25,50,100 µg/mL L. cuneata dichloromethane extract on the protein expression of HO- 1 after treated for 24 h. Western blotting method (treated for 0.5,1,1.5 h)and immunofluorescence staining (treated for 1 h)were used to detect the effects of 100 µg/mL L. cuneata dichloromethane extract on protein expression of Nrf 2 inside and outside the nucleus. After HO-1 gene was silenced by small interfering RNA (Si RNA )transfection technology ,the effects of 100 µg/mL L. cuneata dichloromethane extract on the survival rates of glutamate-induced injury cells and the level of ROS were detected. RESULTS :Compared with blank control ,50, 100 µg/mL L. cuneata dichloromethane extract could significantly improve the survival rate of glutamate-induced injury cells (P< 0.05),while reduced the level of ROS (P<0.05). 25,50, 100 µg/mL L. cuneata dichloromethane extract could increase the protein expression of HO- 1 in cells(P<0.05),while 100 com µg/mL L. cuneata dichloromethane extract could significantly decrease the protein le vel of Nrf 2 in cytoplasm and increasethat in nucleus (P<0.05). After HO-1 gene silencing ,the effects of L. cuneata dichloromethane extract on the proliferation promotion of glutamate-induced injury cells and the reduction of ROS level were reversed (P<0.05). CONCLUSIONS :L. cuneata dichloromethane extract can protect HT 22 cells against injury induced by glutamate through activating Nrf 2 pathway,inducing HO- 1 expression.

3.
Chinese Pharmacological Bulletin ; (12): 1600-1605, 2017.
Article in Chinese | WPRIM | ID: wpr-667307

ABSTRACT

Aim To study the inhibitory effect of volatile components in Oroxyli Semen on liver cancer and its possible mechanisms.Methods H22 bearing mouse model was used,the mice were divided into six groups:blank,model,positive (cytoxan,100 mg · kg-1),low-,mid-,and high-dose (17.5,35,and 70 mg · kg-1) volatile components groups,and then the mice were ig given once daily for consecutive 12 d.Then the tumor growth inhibitory rate,spleen and thymus indexes were calculated;the serum levels of IL-2,IL-6 were determined.HE staining was used to study of the apoptosis of the solid tumor.After treatment of SMMC-7721 cells with 0 ~ 1 g · L-1 of volatile components for 24,48 and 72 h,MTT assay was used to examine the proliferation.TUNEL method was applied to detect cell apoptosis,and RT-PCR method to detect Bax,Bcl-2,caspase-3 mRNA experssion.Results The inhibitory rate of volatile components high-dose on H22 bearing mice was 42.08%.The thymus index and the contents of serum IL-2 and IL-6 of H22 bearing mice were significantly higher than those in model group.Volatile components could significantly inhibit proliferation and induce apoptosis of SMMC-7721 cells,downregulate the expression of Bcl-2 mRNA,and up-regulate the expression of Bax,caspase-3 mRNA.Conclusions The volatile components in Oroxyli Semen have obvious anti-tumor activity in vitro and in vivo,and its mechanism may be related to enhancing immune system and promoting tumor cell apoptosis.

4.
Chinese Pharmacological Bulletin ; (12): 688-691, 2014.
Article in Chinese | WPRIM | ID: wpr-448481

ABSTRACT

Aim To investigate the inhibitory effect of salidroside on tumor growth in tumor-bearing mice and explore its possible mechanisms of anti-tumor action in vivo. Methods The tumor-bearing mice model was established, and the mice were randomly divided into five experimantal groups, including the salidroside low, medium and high doses groups, NS group and positive control matrine group. After continuous injec-tion of drug for eight days, the tumor growth and sur-vival time were measured, tumor growth inhibition rate and life extension rate, spleen index,the content of se-rum IL-2 and IL-12 were calculated. Results The general situation of the salidroside mice was better than the NS group and the positive control matrine group. Compared with NS group, the salidroside low,medium and high doses groups showed inhibitory effects on H22 solid tumor ( P<0. 01 ) with inhibition ratio 28. 9 %, 43. 3 % and 53. 5 %,and lengthened the survival time of mice ( P<0. 01 ) with the life extension rate 37. 54%,48. 96 % and 52. 85 % respectively. The spleen index and the content of serum IL-2 and IL-12 of the salidroside mice were significantly higher than NS group and matrine group. Conclusion Salidroside has marked inhibitory effects on tumor growth in vivo, its mechanism is probably related to improving the anti-tumor immune function.

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