Hypoxia induces apoptosis by caspase activation accompanying cytochrome C release from mitochondria in MC3T3E1 osteoblasts. p38 MAPK is related in hypoxia-induced apoptosis

被引:36
作者
Chae, HJ
Kim, SC
Han, KS
Chae, SW
An, NH
Kim, HM
Kim, HH
Lee, ZH
Kim, HR [1 ]
机构
[1] Wonkwang Univ, Dept Dent Pharmacol, Sch Dent, Iksan 570749, Chonbuk, South Korea
[2] Wonkwang Univ, Wonkwang Dent Res Inst, Sch Dent, Iksan 570749, Chonbuk, South Korea
[3] Chonbuk Natl Univ, Sch Med, Dept Pharmacol, Chonju, South Korea
[4] Chonbuk Natl Univ, Sch Med, Inst Cardiovasc Res, Chonju, South Korea
[5] Wonkwang Univ, Coll Pharm, Sch Dent, Dept Oral Med, Iksan, South Korea
[6] Wonkwang Univ, Dept Orthodont, Iksan, South Korea
[7] Wonkwang Univ, Coll Pharm, Dept Oriental Pharm, Iksan, South Korea
[8] Chosun Univ, Natl Res Lab Bone Metab, Kwangju 501759, South Korea
[9] Chosun Univ, Res Ctr Proteinous Mat, Kwangju 501759, South Korea
关键词
D O I
10.1081/IPH-100103855
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
The aim of this study is to elucidate the possible mechanism of apoptosis in response to hypoxia in MC3T3E1 osteoblasts. MC3T3E1 osteoblasts under hypoxic conditions (2% oxygen) resulted in apoptosis in a time-dependent manner estimated by DNA fragmentation assay and nuclear morphology stained with fluorescent dye, Hoechst 33258. Pretreatment with Z-VAD-FMK, a pan-caspase inhibitor, or Z-DEVD-CHO, a specific caspase-3 inhibitor, completely suppressed the DNA ladder in response to hypoxia. An increase in caspase-3-like protease (DEVDase) activity was observed during apoptosis, but no caspase-1 activity (YVADase) was detected. To confirm what caspases are involved in apoptosis, western blot analysis was performed using anti caspase-3 or -6 antibody. The 10-kDa protein, corresponding to the active products of caspase-3 and the 10-kDA protein of the active protein of caspase-6 were generated in hypoxia-challenged cells in which processing of the full length form of caspase-3 and -6 was evident. With a time course similar to this caspase-3 and -6 activation was evident, hypoxic stress caused the cleavage of lamin A, typical of caspase-6 activity. In addition, the stress elicited the release of cytochrome c into the cytosol during apoptosis. Furthermore, we have observed that pre-treatment with SB203580, a selective p38 MAP kinase (p38 MAPK) inhibitor, attenuated the hypoxia-induced apoptosis. The addition of SB203580 suppressed caspase-3 and -6-like protease activity by hypoxia up to 50%. In contrast, PD98059 had no effect on the hypoxia-induced apoptosis. To confirm the involvement of MAP kinase, JNK/SAPK, ERK, or p38 kinase assay was performed. Although p38 MAPK was activated in response to hypoxic treatment, the other MAP kinase -JNK/SAPK or ERK- was not or modestly activated. These results suggest that p38 MAPK positively regulates hypoxia-induced apoptosis in MC3T3E1 osteoblasts.
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页码:133 / 152
页数:20
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