The objective of study was to investigate the in vitro suppressive effect of angelica polysaccharide (APS) on human cytomegalovirus-induced apoptosis via direct infection in CHRF-288-11 cells. HCMV AD169 directly infected CHRF-288-11 were cultured in vitro, APS in different doses were added on day 3 after the infection of virus. Cells of every group were collected at different time points. HCMV DNA of cells were detected by using polymerase chain reaction and the apoptotic cells were examined by using Hoechst staining, MTT assay, DNA fragmentation assay and flow cytometry. The results showed that the APS to some extent inhibited the apoptosis of CHRF cells infected by HCMV in vitro in a dose-dependent manner. The expression of HCMV IEA in CHRF-288-11 cells was found by PCR amplification. Morphology observation, flow cytometry assay and DNA fragmentation assay revealed the existence of apoptosis. With the dose decrease of APS added to the infected CHRF cells, the percentage of apoptotic cells increased. It is concluded that the HCMV AD169 can infect CHRF cells directly in vitro and decrease cell viability. HCMV AD169 infection increases the apoptosis of CHRF cells in time-dependent manner. When APS was added to the CHRF cells infected by HCMV AD169 in vitro, the viability of CHRF cells increase, which indicated that APS to some extent protects the CHRF cells infected by HCMV. APS suppresses the cytomegalovirus-induced apoptosis in CHRF cells directly infected in vitro in dose-dependent manner.
Angelica ; chemistry ; Apoptosis ; drug effects ; Cells, Cultured ; Cytomegalovirus ; Humans ; Megakaryocytes ; cytology ; drug effects ; virology ; Polysaccharides ; pharmacology

Objective: To reveal the crucial toxic components of ambient fine particles (PM_2.5) that affect the maturation and differentiation of megakaryocytes. Methods: Human megakaryocytes were exposed to the organic fractions, metallic fractions and water-soluble fractions of PM_2.5 at two exposure doses (i.e. actual air proportion concentration or the same concentration), respectively. The cell viability was performed to screen the non-cytotoxic levels of toxic components of PM_2.5 using the CCK-8 assay. CellTiter-Blue assay, morphological observation, flow cytometry analysis and WGA staining assay were used to evaluate the cell morphological changes, occurrence of DNA ploidy, alteration in the expressions of biomarkers and platelet formation, which were key indicators of the maturation and differentiation of megakaryocytes. Results: Compared to the control group, both metallic and organic components of PM_2.5 resulted in a lag in megakaryocytes with an increase in cell volume and the onset of DNA ploidy. Flow cytometry analysis showed that CD33 (the marker of myeloid-specific) decreased and CD41a (a megakaryocyte maturation-associated antigen) increased in metallic and organic components of PM_2.5 treatment groups. Moreover, compared to the control group, budding protrusions increased in metallic and organic components of PM_2.5 treatment groups. The water-soluble components had no effect on the maturation and differentiation of macrophages. Conclusion: Metallic and organic components of PM_2.5 are the crucial toxic components that promote the maturation and differentiation of megakaryocytes.
Biomarkers ; DNA/pharmacology* ; Humans ; Megakaryocytes/chemistry* ; Particulate Matter/toxicity* ; Sincalide/pharmacology* ; Water/pharmacology*

OBJECTIVETo investigate the effect of Angelica polysaccharide (APS), platelet-derived growth factor (PDGF) and thrombopoietin (TPO) on the proliferation and apoptosis of human megakaryocytic cell line M-07e.

METHODSCell count and the viability testing of M-07e cells (trypan blue exclusion assay) were performed at 24 hours, 48 hours and 72 hours after treatment with APS, PDGF or TPO. Three apoptosis related flow cytometric assays including Annexin V, Caspase-3 and JC-1 were performed to determine apoptotic rate of each group at 72 hours after the treatment.

RESULTSAfter the incubation, the number of M-07e cells in the APS, PDGF and TPO group increased and the viabilities of the three groups were significantly higher than the control group (P < 0.05). The dead cells in the APS, PDGF and TPO group were (19.41 +/- 7.59)%, (21.38 +/- 7.25)% and (18.77 +/- 8.00)%, respectively by flow cytometry using Annexin V method, which were significantly lower compared to the control group (34.33 +/- 5.46)%. The expression of the activated caspase-3 in the group of APS, PDGF and TPO were (12.27 +/- 5.18)%, (12.39 +/- 6.26)% and (13.75 +/- 8.25)%, the APS and PDGF group decreased significantly compared to the control group (18.92 +/- 6.09)%. The ratio of total cell deaths in the APS, PDGF and TPO group were (23.64 +/- 6.69)%, (28.00 +/- 10.05)% and (27.99 +/- 8.99)%, the ratio in APS group decreased significantly compared to the control group (39.48 +/- 11.86)% by JC-1 method. Differences between APS and PDGF groups and between APS and TPO groups were not statistically significant.

CONCLUSIONAPS, PDGF and TPO have similar effect in stimulating proliferation and inhibiting serum-free-culture induced apoptosis of M-07e cells.

Angelica ; chemistry ; Apoptosis ; drug effects ; Benzimidazoles ; pharmacology ; Carbocyanines ; pharmacology ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Flow Cytometry ; Fluorescent Dyes ; pharmacology ; Humans ; Megakaryocytes ; drug effects ; physiology ; Organic Chemicals ; pharmacology ; Platelet-Derived Growth Factor ; pharmacology ; Thrombopoiesis ; Thrombopoietin ; pharmacology