Increased circulating heat shock protein 60 induced by menopause, stimulates apoptosis of osteoblast-lineage cells via up-regulation of toll-like receptors.
Postmenopausal osteoporosis is a heterogeneous disorder characterized by accelerated bone loss after natural or surgical menopause and an increased risk of fractures. The bone loss in estrogen deficiency results from the increased bone resorption and impaired ability of osteoblastic bone formation. Previous studies have reported that the HSP60 stimulates osteoclast formation and bone resorption. Here we found that plasma HSP60 levels were significantly higher in postmenopausal (median 1152.4 ng/ml; range 724.7-2123.4 ng/ml) than in premenopausal (median 316.3 ng/ml; range 164.6-638.4 ng/ml) women. In primary human bone marrow stromal cells (hBMSC) and the HS-5 hBMSC cell line, HSP60 significantly reduced cell viability and increased caspase-dependent apoptosis. Consistent with these observations, HSP60 activated caspase-3 and -9, but not caspase-8 in HS-5 cells, and increased the release of mitochondrial cytochrome c into the cytosol. In addition, HSP60 activated p38 and NFkappaB, but not ERK or JNK; importantly, inhibitors of p38 (SB203580) and NFkappaB (PDTC) abolished HSP60-induced apoptosis. Furthermore, Western blotting showed that HSP60 up-regulated TLR-2 and TLR-4 expression, and pretreatment with blocking antibodies for TLR-2 and TLR-4 almost completely eliminated the effects of HSP60 on apoptosis, caspase-3 and -9 activation, and activation of NFkappaB and p38 MAPK. Most notably, ovariectomy-induced bone loss was attenuated in TLR-2 KO mice. In conclusion, up-regulation of TLR-2 by HSP60 may play a critical role in promoting bone loss in the estrogen-deficient state.
Kim
Yang Soon
YS
Asan Institute for Life Sciences, Seoul, Republic of Korea.
Koh
Jung-Min
JM
Lee
Young-Sun
YS
Kim
Beom-Jun
BJ
Lee
Seung Hun
SH
Lee
Ki-Up
KU
Kim
Ghi Su
GS
D000328:Adult; D000818:Animals; D017209:Apoptosis; D001862:Bone Resorption / Q000378:metabolism / Q000473:pathology; D020169:Caspases / Q000378:metabolism; D019070:Cell Lineage; D002470:Cell Survival; D018834:Chaperonin 60 / Q000097:blood*; D005260:Female; D006801:Humans; D008593:Menopause / Q000097:blood*; D051379:Mice; D018345:Mice, Knockout; D008875:Middle Aged; D008928:Mitochondria / Q000201:enzymology; D010006:Osteoblasts / Q000166:cytology* / Q000201:enzymology; D010052:Ovariectomy; D017698:Postmenopause / Q000097:blood; D017697:Premenopause / Q000097:blood; D051195:Toll-Like Receptor 2 / Q000235:genetics / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000378:metabolism; D051193:Toll-Like Receptors / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation
D018834:Chaperonin 60; C495342:TLR2 protein, human; C495345:TLR4 protein, human; C493486:Tlr2 protein, mouse; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors; D020169:Caspases