Two new ursane triterpenoids from Agastache rugosa (Fisch. et.Mey.) O. Kuntze / 药学学报

Two new ursane triterpenoids along with twelve known compounds were isolated from 80% ethanol extract of Agastache rugosa (Fisch. et. Mey.) O. Kuntze by using silica gel column, MCI column, ODS column and HPLC. The structures of the new compounds were identified as 2α,3α-dihydroxy-24-nor-urs-4(23),12(13)-dien-28-oic acid (1) and 2α,3α-dihydroxy-24-nor-urs-4(23),12(13),20(30) -trien-28-oic acid (2) by HR-ESI-MS, NMR and ECD spectral data, named agasursacid A and agasursacid B. In addition, compounds 3, 4, 6, 8 showed anti-coxsackievirus B3 (CVB3) activities with a IC50 as 4.77, 1.59, 11.11 and 25.87 μmol·L-1, resepectively.

Coxsackievirus B3 Infection Triggers Autophagy through 3 Pathways of Endoplasmic Reticulum Stress / 生物医学与环境科学（英文）

OBJECTIVE@#Autophagy is a highly conserved intracellular degradation pathway. Many picornaviruses induce autophagy to benefit viral replication, but an understanding of how autophagy occurs remains incomplete. In this study, we explored whether coxsackievirus B3 (CVB3) infection induced autophagy through endoplasmic reticulum (ER) stress.@*METHODS@#In CVB3-infected HeLa cells, the specific molecules of ER stress and autophagy were detected using Western blotting, reverse transcription polymerase chain reaction (RT-PCR), and confocal microscopy. Then PKR-like ER protein kinase (PERK) inhibitor, inositol-requiring protein-1 (IRE1) inhibitor, or activating transcription factor-6 (ATF6) inhibitor worked on CVB3-infected cells, their effect on autophagy was assessed by Western blotting for detecting microtubule-associated protein light chain 3 (LC3).@*RESULTS@#CVB3 infection induced ER stress, and ER stress sensors PERK/eIF2α, IRE1/XBP1, and ATF6 were activated. CVB3 infection increased the accumulation of green fluorescent protein (GFP)-LC3 punctuation and induced the conversion from LC3-I to phosphatidylethanolamine-conjugated LC3-1 (LC3-II). CVB3 infection still decreased the expression of mammalian target of rapamycin (mTOR) and p-mTOR. Inhibition of PERK, IRE1, or ATF6 significantly decreased the ratio of LC3-II to LC3-I in CVB3-infected HeLa cells.@*CONCLUSION@#CVB3 infection induced autophagy through ER stress in HeLa cells, and PERK, IRE1, and ATF6a pathways participated in the regulation of autophagy. Our data suggested that ER stress may inhibit mTOR signaling pathway to induce autophagy during CVB3 infection.

Expression of coxsackievirus group B3 gene fragment encoding VP1 in procaryon and clinical significance / 中国免疫学杂志

Objective:To study the expression of coxsackievirus group B3 gene fragment encoding VP1 in procaryon and to explore its application.Methods:Stablie expression of VP1 gene of CVB3 in E.coli was obtained.The expressed protein was purified by NAT chromatography and its immunoactivity was identified by indirect ELISA.Results:The expressed product of VP1,similar to native protein antigen of CVB3,could strong bind with the mouse's antibody serum against CVB3(polyclonal antibody).Irrelevant monoclonal antibody as contrast presents negative activity. Using the expressed VP1 product,we have had a special IgM ELISA for the patient's serum of the clinical acute viral myocarditis.The result was same with the cellular protein antigens of the tissue-cultivated CVB3.Conclusion:The protein antigen CVB3-VP1 which is obtained by the method of gene engineering has character of high product, and its immunoactivity after being purified was basically unchanged. At present, this kind of antigen is difficult to be obtained from the viral cullture medium and a potent hazard for being infected by this virus may take place in such manipulateion. By the method of gene engineering we can obtain antigenic VP1 of CVB3 and use as immuneogen for the detection of serum antibody,by which to provide reliable test reference for the early-stage diagnosis and clinical therapy of the acute myocarditis.