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Protective Effect and Mechanism of Ginkgolide B on Amyotrophic Lateral Sclerosis Cell Model / 中国实验方剂学杂志
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 33-38, 2020.
Article in Chinese | WPRIM | ID: wpr-873016
ABSTRACT

Objective:

To investigate the effect of ginkgolide B (GB) on the activation of c-Jun aminoterminal kinase(JNK) signaling pathway and apoptosis in amyotrophic lateral sclerosis cell model.

Method:

NSC34 cells were infected by slow virus containing expression superoxide dismutase1(SOD1)WT and hSOD1G93A and empty plasmid, and screened with a certain concentration of puromycin, so as to observe the transfection efficiency of slow virus and cell morphology under inverted fluorescence microscope. Western blot method was used to verify whether infected cells were over-expressing SOD1 target proteins. The hSOD1G93A-NSC34 cell lines were established and given GB. Cell cultures were divided into normal group, model group and different concentrations of ginkgolide B groups (25, 50, 75, 100 mg∙L-1). After 48 h, methyl thiazolyl tetrazolium (MTT) was used to detect cell survival rates, and select the best drug concentration. Subsequent experimental groups were divided into normal group, model group, 75 mg∙L-1 GB group, SP600125 group, and 75 mg∙L-1 GB + SP600125 group. Flow cytometry was used to detect the apoptosis of each group of cells. Western blot was used to detect the expressions of phosphorylation(p)-JNK, c-Jun, p-c-Jun, and cysteine aspartic acid protease -3(Caspase-3proteins.

Result:

Compared with normal NSC34 cells, hSOD1G93A-NSC34 cell body became round, the synapses decreased and shortened, but the cell morphology of hSODWT-NSC34 cell and empty plasmid group did not change significantly. Western blot showed that hSOD1G93A-NSC34, hSOD1WT-NSC3 intracellular SOD1 protein levels increased significantly (P<0.01), and the amyotrophic lateral sclerosis cell model was established. Compared with the normal group, the cell activity in the model group was significantly reduced (P<0.01). Compared with the model group, the cell activity increased at different concentrations of GB, especially when the drug concentration was 75 mg∙L-1 (P<0.01). In subsequent experiments, compared with the normal group, the apoptosis, and expressions of p-JNK, p-c-Jun, and cleaved Caspase-3 proteins in the model group increased significantly (P<0.01). Compared with the model group, the apoptosis and p-JNK, p-c-Jun, released Caspase-3 protein expressions of 75 mg∙L-1 GB group, SP600125 group, 75 mg∙L-1 GB + SP600125 group decreased significantly (P<0.05, P<0.01).

Conclusion:

GB has a protective effect on the cell model of atrophy lateral sclerosis, which may be realized by JNK signal pathway.

Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2020 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2020 Type: Article