Objective The present study aims to investigate the role of protein kinase C 5 subtype （PKCS） phosphorylation in the process of 6-hydroxydopamine （6-OHDA）-induced dopaminergic cell death, and demonstrate the molec...Objective The present study aims to investigate the role of protein kinase C 5 subtype （PKCS） phosphorylation in the process of 6-hydroxydopamine （6-OHDA）-induced dopaminergic cell death, and demonstrate the molecular basis of neurological disorders, such as Parkinson＇ s disease. Methods The pheochromocytoma （PC 12） cell line was employed in the present study. Cells were treated with 2 μmol/L PKC5 inhibitor Rottlerin, 10 nmol/L protein kinase C α subtype （PKCα） inhibitor bisindolylmaleimide 1, or 5 nmol/L G66976 that could specifically inhibit the calcium-dependent PKC isoforms, respectively. PKC8 activator phorbol-12-myristate-13-acetate （PMA, 100 nmol/L） was also used in this study. All these agents were added to the medium before cells were incubated with 6-OHDA. Cells with no treatment served as control. The cytotox- icity of 6-OHDA was determined by methyl thiazolyl tetrazolium （MTT） reduction assay and PKCδ phosphorylation levels in various groups were measured by western blotting. Results Bisindolylmaleimide I and Go6976 exerted no significant attenuation on the cytotoxicity of 6-OHDA, nor any effects on PKCδ phosphorylation in PC 12 cells. However, Rottlerin could inhibit the phosphorylation of PKC5 and attenuate 6-OHDA-induced cell death, and the cell viability was raised to 69.6 ±2.63% of that in control group （P 〈 0.05）. In contrast, PMA induced a significant increase in PKC5 phosphorylation and also strengthened the cytotoxic effects of 6-OHDA. The cell viability of PMA-treated PC12 cells decreased to 49.8±5.06% of that in control group （P 〈 0.001）. Conclusion Rottlerin can protect PC 12 cells from cytotoxicity of 6-OHDA probably by inhibiting PKC δ phosphorylation. The results suggest that PKCδ may be a key regulator of neuron loss in Parkinson＇s disease.展开更多
Infection of Zika virus(ZIKV)may cause microcephaly and other neurological disorders,while no vaccines and drugs are available.Our study revealed that rottlerin confers a broad antiviral activity against several envel...Infection of Zika virus(ZIKV)may cause microcephaly and other neurological disorders,while no vaccines and drugs are available.Our study revealed that rottlerin confers a broad antiviral activity against several enveloped viruses,including ZIKV,vesicular stomatitis virus,and herpes simplex virus,but not against two naked viruses(enterovirus 71 and encephalomyocarditis virus).Rottlerin does not have a direct virucidal effect on the virions,and its antiviral effect is independent of its regulation on PKCδor ATP.Both pretreatment and post-treatment of rottlerin effectively reduce the viral replication of ZIKV.The pretreatment of rottlerin disturbs the endocytosis of enveloped viruses,while the post-treatment of rottlerin acts at a late stage through disturbing the maturation of ZIKV.Importantly,administration of rottlerin in neonatal mice significantly decreased the ZIKV replication in vivo,and alleviated the neurological symptoms caused by ZIKV.Our work suggests that rottlerin exerts an antiviral activity at two distinct steps of viral infection,and can be potentially developed as a prophylactic and therapeutic agent.展开更多
Vidangadi churna is a popular Ayurvedic formulation described in the chapter Krimicikitsa of the Ayurvedic literature Cakradatta for the treatment of Krimiroga. The preparation is a composite mixture of the fine powde...Vidangadi churna is a popular Ayurvedic formulation described in the chapter Krimicikitsa of the Ayurvedic literature Cakradatta for the treatment of Krimiroga. The preparation is a composite mixture of the fine powder of fruits of Vidang （Embelia ribs）, glandular trichomes of the fruits of Kamala （Mallotus philippensis）, mature fruits of Harde （Terminalia chehula）, Saindhava and Yavakshara. The use of reversed phase C18 column eluted with gradient mobile phase of acetonitrile and water enabled the efficient separation of the chemical markers in 22 rain. Validation of the method was performed in order to demonstrate its selectivity, accuracy, precision, repeatability and recovery. All calibration curves showed good linear correlation coefficients （r2〉0.995） within the tested ranges. Three markers in Vidangadi churna were quantitied with respect to Embelin （0.647%, w/w）, Rottlerin （4.419%, w/w）, and Ellagic acid （0.459%, w/w）. lntraand inter-day RSDs of retention times and peak areas were less than 3.12%. The recoveries were between 99.66% and 102.33%. In conclusion, a method has been developed for the simultaneous quantification of three markers in Vidangadi churna. The RP-HPLC method was simple, precise and accurate and can be used for the quality control of the raw materials as well as formulations.展开更多
基金supported by the International Cooperation Project of Science and Technology Department of Heilongjiang Province, China (No.WB04301, No.WB08B05)the Science and Technology Foundation of Education Department of Heilongjiang Province, China (No.11521076)
文摘Objective The present study aims to investigate the role of protein kinase C 5 subtype （PKCS） phosphorylation in the process of 6-hydroxydopamine （6-OHDA）-induced dopaminergic cell death, and demonstrate the molecular basis of neurological disorders, such as Parkinson＇ s disease. Methods The pheochromocytoma （PC 12） cell line was employed in the present study. Cells were treated with 2 μmol/L PKC5 inhibitor Rottlerin, 10 nmol/L protein kinase C α subtype （PKCα） inhibitor bisindolylmaleimide 1, or 5 nmol/L G66976 that could specifically inhibit the calcium-dependent PKC isoforms, respectively. PKC8 activator phorbol-12-myristate-13-acetate （PMA, 100 nmol/L） was also used in this study. All these agents were added to the medium before cells were incubated with 6-OHDA. Cells with no treatment served as control. The cytotox- icity of 6-OHDA was determined by methyl thiazolyl tetrazolium （MTT） reduction assay and PKCδ phosphorylation levels in various groups were measured by western blotting. Results Bisindolylmaleimide I and Go6976 exerted no significant attenuation on the cytotoxicity of 6-OHDA, nor any effects on PKCδ phosphorylation in PC 12 cells. However, Rottlerin could inhibit the phosphorylation of PKC5 and attenuate 6-OHDA-induced cell death, and the cell viability was raised to 69.6 ±2.63% of that in control group （P 〈 0.05）. In contrast, PMA induced a significant increase in PKC5 phosphorylation and also strengthened the cytotoxic effects of 6-OHDA. The cell viability of PMA-treated PC12 cells decreased to 49.8±5.06% of that in control group （P 〈 0.001）. Conclusion Rottlerin can protect PC 12 cells from cytotoxicity of 6-OHDA probably by inhibiting PKC δ phosphorylation. The results suggest that PKCδ may be a key regulator of neuron loss in Parkinson＇s disease.
基金supported by Guangzhou Municipal Science and Technology Program (202206010114)National Natural Science Foundation of China (31970887, 92169110)+1 种基金Doctoral workstation foundation of Guangdong Second Provincial General hospital (2020BSGZ036)Natural Science Foundation of Guangdong Province(2021A1515011491, 2022A1515010451)
文摘Infection of Zika virus(ZIKV)may cause microcephaly and other neurological disorders,while no vaccines and drugs are available.Our study revealed that rottlerin confers a broad antiviral activity against several enveloped viruses,including ZIKV,vesicular stomatitis virus,and herpes simplex virus,but not against two naked viruses(enterovirus 71 and encephalomyocarditis virus).Rottlerin does not have a direct virucidal effect on the virions,and its antiviral effect is independent of its regulation on PKCδor ATP.Both pretreatment and post-treatment of rottlerin effectively reduce the viral replication of ZIKV.The pretreatment of rottlerin disturbs the endocytosis of enveloped viruses,while the post-treatment of rottlerin acts at a late stage through disturbing the maturation of ZIKV.Importantly,administration of rottlerin in neonatal mice significantly decreased the ZIKV replication in vivo,and alleviated the neurological symptoms caused by ZIKV.Our work suggests that rottlerin exerts an antiviral activity at two distinct steps of viral infection,and can be potentially developed as a prophylactic and therapeutic agent.
文摘Vidangadi churna is a popular Ayurvedic formulation described in the chapter Krimicikitsa of the Ayurvedic literature Cakradatta for the treatment of Krimiroga. The preparation is a composite mixture of the fine powder of fruits of Vidang （Embelia ribs）, glandular trichomes of the fruits of Kamala （Mallotus philippensis）, mature fruits of Harde （Terminalia chehula）, Saindhava and Yavakshara. The use of reversed phase C18 column eluted with gradient mobile phase of acetonitrile and water enabled the efficient separation of the chemical markers in 22 rain. Validation of the method was performed in order to demonstrate its selectivity, accuracy, precision, repeatability and recovery. All calibration curves showed good linear correlation coefficients （r2〉0.995） within the tested ranges. Three markers in Vidangadi churna were quantitied with respect to Embelin （0.647%, w/w）, Rottlerin （4.419%, w/w）, and Ellagic acid （0.459%, w/w）. lntraand inter-day RSDs of retention times and peak areas were less than 3.12%. The recoveries were between 99.66% and 102.33%. In conclusion, a method has been developed for the simultaneous quantification of three markers in Vidangadi churna. The RP-HPLC method was simple, precise and accurate and can be used for the quality control of the raw materials as well as formulations.
