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Apoptogenic activity of 2α,3α-dihydroxyurs-12-ene-28-oic acid from Prunella vulgaris var. lilacina is mediated via mitochondria-dependent activation of caspase cascade regulated by Bcl-2 in human acute leukemia Jurkat T cells.
J Ethnopharmacol. 2011 Jun 01; 135(3):626-35.JE

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

The dried spikes of Prunella vulgaris var. lilacina (Labiatae) have been used for traditional herbal medicine to treat fever, inflammation, dropsy, gonorrhea and cancer in Korea, Japan and China. The present study evaluated the apoptotic effect of 2α,3α-dihydroxyurs-12-en-28-oic acid (DHURS), purified from the dried spikes on human acute leukemia Jurkat T cells.

MATERIALS AND METHODS

Cell viability was assessed by MTT assay. Mitochondrial membrane potential (Δψm) loss, apoptotic change of the cell cycle, and apoptotic cells were measured by flow cytometric analysis. Mitochondrial cytochrome c release and activation of caspase cascade were determined by Western blot analysis. Caspase-12 activity and caspase-3 activity were assayed using the Fluorometric Assay Kit and the Colorimetric Assay Kit, respectively.

RESULTS

Treatment of Jurkat T cells with DHURS (20-25 μg/ml) caused cytotoxicity and apoptotic DNA fragmentation along with Δψm loss, mitochondrial cytochrome c release, activation of caspase-9, -7, -3, and -8, and PARP degradation. However, these apoptotic events were abrogated by overexpression of Bcl-2. Pretreatment of the cells with the pan-caspase inhibitor (z-VAD-fmk), the caspase-9 inhibitor (z-LEHD-fmk) or the caspase-3 inhibitor (z-DEVD-fmk) to prevent DHURS-induced apoptosis could block the activation of caspase-7 and -8, and PARP degradation, but not the Δψm loss, activation of caspase-9 and -3. Both FADD- and caspase-8-positive wild-type Jurkat clone A3, FADD-deficient Jurkat clone I2.1, and caspase-8-deficient Jurkat clone I9.2 exhibited similar susceptibilities to the cytotoxicity of DHURS, excluding an involvement of Fas/FasL system in triggering the apoptosis. The IC(50) value for Jurkat T cells was ∼22 μg/ml, whereas that for human peripheral T cells was 25 μg/ml.

CONCLUSIONS

These results indicate that DHURS-induced apoptogenic activity in Jurkat T cells, which was less potent in normal peripheral T cells, was mediated by Δψm loss, mitochondrial cytochrome c release, and subsequent activation of caspase-9 and -3, leading to activation of caspase-7 and -8, which could be regulated by Bcl-2.

Authors+Show Affiliations

Laboratory of Immunobiology, School of Life Science and Biotechnology, College of Natural Sciences, Kyungpook National University, Daegu, Republic of Korea.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21473903

Citation

Woo, Hyun Ju, et al. "Apoptogenic Activity of 2α,3α-dihydroxyurs-12-ene-28-oic Acid From Prunella Vulgaris Var. Lilacina Is Mediated Via Mitochondria-dependent Activation of Caspase Cascade Regulated By Bcl-2 in Human Acute Leukemia Jurkat T Cells." Journal of Ethnopharmacology, vol. 135, no. 3, 2011, pp. 626-35.
Woo HJ, Jun do Y, Lee JY, et al. Apoptogenic activity of 2α,3α-dihydroxyurs-12-ene-28-oic acid from Prunella vulgaris var. lilacina is mediated via mitochondria-dependent activation of caspase cascade regulated by Bcl-2 in human acute leukemia Jurkat T cells. J Ethnopharmacol. 2011;135(3):626-35.
Woo, H. J., Jun, d. o. . Y., Lee, J. Y., Woo, M. H., Yang, C. H., & Kim, Y. H. (2011). Apoptogenic activity of 2α,3α-dihydroxyurs-12-ene-28-oic acid from Prunella vulgaris var. lilacina is mediated via mitochondria-dependent activation of caspase cascade regulated by Bcl-2 in human acute leukemia Jurkat T cells. Journal of Ethnopharmacology, 135(3), 626-35. https://doi.org/10.1016/j.jep.2011.03.067
Woo HJ, et al. Apoptogenic Activity of 2α,3α-dihydroxyurs-12-ene-28-oic Acid From Prunella Vulgaris Var. Lilacina Is Mediated Via Mitochondria-dependent Activation of Caspase Cascade Regulated By Bcl-2 in Human Acute Leukemia Jurkat T Cells. J Ethnopharmacol. 2011 Jun 1;135(3):626-35. PubMed PMID: 21473903.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Apoptogenic activity of 2α,3α-dihydroxyurs-12-ene-28-oic acid from Prunella vulgaris var. lilacina is mediated via mitochondria-dependent activation of caspase cascade regulated by Bcl-2 in human acute leukemia Jurkat T cells. AU - Woo,Hyun Ju, AU - Jun,Do Youn, AU - Lee,Ji Young, AU - Woo,Mi Hee, AU - Yang,Chae Ha, AU - Kim,Young Ho, Y1 - 2011/04/05/ PY - 2010/12/14/received PY - 2011/02/17/revised PY - 2011/03/28/accepted PY - 2011/4/9/entrez PY - 2011/4/9/pubmed PY - 2011/12/30/medline SP - 626 EP - 35 JF - Journal of ethnopharmacology JO - J Ethnopharmacol VL - 135 IS - 3 N2 - ETHNOPHARMACOLOGICAL RELEVANCE: The dried spikes of Prunella vulgaris var. lilacina (Labiatae) have been used for traditional herbal medicine to treat fever, inflammation, dropsy, gonorrhea and cancer in Korea, Japan and China. The present study evaluated the apoptotic effect of 2α,3α-dihydroxyurs-12-en-28-oic acid (DHURS), purified from the dried spikes on human acute leukemia Jurkat T cells. MATERIALS AND METHODS: Cell viability was assessed by MTT assay. Mitochondrial membrane potential (Δψm) loss, apoptotic change of the cell cycle, and apoptotic cells were measured by flow cytometric analysis. Mitochondrial cytochrome c release and activation of caspase cascade were determined by Western blot analysis. Caspase-12 activity and caspase-3 activity were assayed using the Fluorometric Assay Kit and the Colorimetric Assay Kit, respectively. RESULTS: Treatment of Jurkat T cells with DHURS (20-25 μg/ml) caused cytotoxicity and apoptotic DNA fragmentation along with Δψm loss, mitochondrial cytochrome c release, activation of caspase-9, -7, -3, and -8, and PARP degradation. However, these apoptotic events were abrogated by overexpression of Bcl-2. Pretreatment of the cells with the pan-caspase inhibitor (z-VAD-fmk), the caspase-9 inhibitor (z-LEHD-fmk) or the caspase-3 inhibitor (z-DEVD-fmk) to prevent DHURS-induced apoptosis could block the activation of caspase-7 and -8, and PARP degradation, but not the Δψm loss, activation of caspase-9 and -3. Both FADD- and caspase-8-positive wild-type Jurkat clone A3, FADD-deficient Jurkat clone I2.1, and caspase-8-deficient Jurkat clone I9.2 exhibited similar susceptibilities to the cytotoxicity of DHURS, excluding an involvement of Fas/FasL system in triggering the apoptosis. The IC(50) value for Jurkat T cells was ∼22 μg/ml, whereas that for human peripheral T cells was 25 μg/ml. CONCLUSIONS: These results indicate that DHURS-induced apoptogenic activity in Jurkat T cells, which was less potent in normal peripheral T cells, was mediated by Δψm loss, mitochondrial cytochrome c release, and subsequent activation of caspase-9 and -3, leading to activation of caspase-7 and -8, which could be regulated by Bcl-2. SN - 1872-7573 UR - https://www.unboundmedicine.com/medline/citation/21473903/Apoptogenic_activity_of_2α3α_dihydroxyurs_12_ene_28_oic_acid_from_Prunella_vulgaris_var__lilacina_is_mediated_via_mitochondria_dependent_activation_of_caspase_cascade_regulated_by_Bcl_2_in_human_acute_leukemia_Jurkat_T_cells_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0378-8741(11)00218-2 DB - PRIME DP - Unbound Medicine ER -