Apoptosis of bone marrow-derived mesenchymal stem cells (BMMSCs) is an essential pathogenic factor of osteoporosis. Ginsenoside-Rb2 (Rb2), a 20(S)-protopanaxadiol glycoside extracted from ginseng, is a potent treatment for bone loss, which raises interest regarding the bone metabolism area. In the present study, we found that dose–response Rb2 inhibited high dosage of dexamethasone (Dex)-induced apoptosis in primary murine BMMSCs. Interestingly, Rb2 promoted GPR120 induction, which is the unsaturated long-chain fatty acid receptor. We further confirmed that GPR120-specific ShRNA reversed the inhibition of Rb2 on Dex-induced apoptosis by activating caspase-3 and reducing cell viability. In addition, Rb2 notably increased phosphorylated ERK1/2 levels and Ras kinase activity dependently through the GPR120. The ERK1/2 activity-specific inhibitor U0126 remarkably blocked the Rb2-induced antiapoptotic effect in response to Dex-induced apoptosis. Together, dose–response Rb2 protected BMMSCs against Dex-induced apoptosis dependently by inducing GPR120 promoted Ras-ERK1/2 signaling pathway. Therefore, in the prevalence of the abuse of Dex in the clinic, our findings suggest for the first time that Rb2 is not only a key to understand the link between Chinese medicine and the pathology of osteoporosis but also an underlying target for the treatment of bone complications in the foreseeable future.
Get full access to this article
View all access options for this article.
References
1.
RodanGA and MartinTJ. (2000). Therapeutic approaches to bone diseases. Science, 289:1508–1514.
2.
KarsentyG and WagnerEF. (2002). Reaching a genetic and molecular understanding of skeletal development. Dev Cell, 2:389–406.
3.
RaiszLG. (2005). Pathogenesis of osteoporosis: concepts, conflicts, and prospects. J Clin Invest, 115:3318–3325.
4.
ChenTL. (2004). Inhibition of growth and differentiation of osteoprogenitors in mouse bone marrow stromal cell cultures by increased donor age and glucocorticoid treatment. Bone, 35:83–95.
5.
YamazaT, MiuraY, BiY, LiuY, AkiyamaK, SonoyamaW, PatelV, GutkindS, YoungM, et al. (2008). Pharmacologic stem cell based intervention as a new approach to osteoporosis treatment in rodents. PLoS One, 3:e2615
6.
OzHS and HughesWT. (1997). Novel anti-Pneumocystis carinii effects of the immunosuppressant mycophenolate mofetil in contrast to provocative effects of tacrolimus, sirolimus, and dexamethasone. J Infect Dis, 175:901–904.
7.
CollFM, ShuvaevVV, ZernBJ, CompostoRJ, MuzykantovVR and EckmannDM. (2014). Icam-1 targeted nanogels loaded with dexamethasone alleviate pulmonary inflammation. PLoS One, 9:e102329.
8.
MedradoGC, MachadoCB, ValerioP, SanchesMD and GoesAM. (2006). The effect of a chitosan-gelatin matrix and dexamethasone on the behavior of rabbit mesenchymal stem cells. Biomed Mater, 1:155–161.
9.
WangH, PangB, LiY, ZhuD, PangT and LiuY. (2012). Dexamethasone has variable effects on mesenchymal stromal cells. Cytotherapy, 14:423–430.
10.
MuragliaA, CanceddaR and QuartoR. (2000). Clonal mesenchymal progenitors from human bone marrow differentiate in vitro according to a hierarchical model. J Cell Sci, 113 (Pt 7):1161–1166.
11.
HongD, ChenHX, XueY, LiDM, WanXC, GeR and LiJC. (2009). Osteoblastogenic effects of dexamethasone through upregulation of TAZ expression in rat mesenchymal stem cells. J Steroid Biochem Mol Biol, 116:86–92.
12.
SongIH, CaplanAI and DennisJE. (2009). Dexamethasone inhibition of confluence-induced apoptosis in human mesenchymal stem cells. J Orthop Res, 27:216–221.
13.
LeeKT, JungTW, LeeHJ, KimSG, ShinYS and WhangWK. (2011). The antidiabetic effect of ginsenoside Rb2 via activation of AMPK. Arch Pharm Res, 34:1201–1208.
14.
FujimotoJ, SakaguchiH, AokiI, ToyokiH, KhatunS and TamayaT. (2001). Inhibitory effect of ginsenoside-Rb2 on invasiveness of uterine endometrial cancer cells to the basement membrane. Eur J Gynaecol Oncol, 22:339–341.
15.
KangKS, KimHY, BaekSH, YooHH, ParkJH and YokozawaT. (2007). Study on the hydroxyl radical scavenging activity changes of ginseng and ginsenoside-Rb2 by heat processing. Biol Pharm Bull, 30:724–728.
16.
KimEJ, LeeHI, ChungKJ, NohYH, RoY and KooJH. (2009). The ginsenoside-Rb2 lowers cholesterol and triacylglycerol levels in 3T3-L1 adipocytes cultured under high cholesterol or fatty acids conditions. BMB Rep, 42:194–199.
17.
YoshikawaM, MorikawaT, YashiroK, MurakamiT and MatsudaH. (2001). Bioactive saponins and glycosides. XIX. Notoginseng (3): immunological adjuvant activity of notoginsenosides and related saponins: structures of notoginsenosides-L, -M, and -N from the roots of Panax notoginseng (Burk.) F. H. Chen. Chem Pharm Bull (Tokyo), 49:1452–1456.
18.
SunHX, QinF and YeYP. (2005). Relationship between haemolytic and adjuvant activity and structure of protopanaxadiol-type saponins from the roots of Panax notoginseng. Vaccine, 23:5533–5542.
19.
SunH, YangZ and YeY. (2006). Structure and biological activity of protopanaxatriol-type saponins from the roots of Panax notoginseng. Int Immunopharmacol, 6:14–25.
20.
ZhangY, HanLF, SakahKJ, WuZZ, LiuLL, AgyemangK, GaoXM and WangT. (2013). Bioactive protopanaxatriol type saponins isolated from the roots of Panax notoginseng (Burk.) F. H. Chen. Molecules, 18:10352–10366.
21.
SatoK, MochizukiM, SaikiI, YooYC, SamukawaK and AzumaI. (1994). Inhibition of tumor angiogenesis and metastasis by a saponin of Panax ginseng, ginsenoside-Rb2. Biol Pharm Bull, 17:635–639.
22.
ChoiS. (2002). Epidermis proliferative effect of the Panax ginseng ginsenoside Rb2. Arch Pharm Res, 25:71–76.
23.
HuangQ, GaoB, JieQ, WeiBY, FanJ, ZhangHY, ZhangJK, LiXJ, ShiJ, et al. (2014). Ginsenoside-Rb displays anti-osteoporosis effects through reducing oxidative damage and bone-resorbing cytokines during osteogenesis. Bone, 66:306–314.
24.
LiuY, ChenLY, SokolowskaM, EberleinM, AlsaatyS, Martinez-AntonA, LogunC, QiHY and ShelhamerJH. (2014). The fish oil ingredient, docosahexaenoic acid, activates cytosolic phospholipase A via GPR120 receptor to produce prostaglandin E and plays an anti-inflammatory role in macrophages. Immunology, 143:81–95.
25.
KatsumaS, HataeN, YanoT, RuikeY, KimuraM, HirasawaA and TsujimotoG. (2005). Free fatty acids inhibit serum deprivation-induced apoptosis through GPR120 in a murine enteroendocrine cell line STC-1. J Biol Chem, 280:19507–19515.
26.
BouffiC, BonyC, CourtiesG, JorgensenC and NoelD. (2010). IL-6-dependent PGE2 secretion by mesenchymal stem cells inhibits local inflammation in experimental arthritis. PLoS One, 5:e14247.
27.
DominiciM, Le BlancK, MuellerI, Slaper-CortenbachI, MariniF, KrauseD, DeansR, KeatingA, ProckopD and HorwitzE. (2006). Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8:315–317.
28.
ZhangJK, YangL, MengGL, YuanZ, FanJ, LiD, ChenJZ, ShiTY, HuHM, et al. (2013). Protection by salidroside against bone loss via inhibition of oxidative stress and bone-resorbing mediators. PLoS One, 8:e57251
29.
ZhangZ, LiuW, ZhengY, ZhuR, ZhuY, YaoW and GaoX. (2014). The ERK/eIF4F/Bcl-XL pathway mediates SGP-2 induced osteosarcoma cells apoptosis in vitro and in vivo. Cancer Lett, 352:203–213.
30.
ZhengJ, DuW, SongLJ, ZhangR, SunLG, ChenFG and WeiXT. (2014). Norcantharidin induces growth inhibition and apoptosis of glioma cells by blocking the Raf/MEK/ERK pathway. World J Surg Oncol, 12:207.
31.
WangB, QinH, WangY, ChenW, LuoJ, ZhuX, WenW and LeiW. (2014). Effect of DJ-1 overexpression on the proliferation, apoptosis, invasion and migration of laryngeal squamous cell carcinoma SNU-46 cells through PI3K/AKT/mTOR. Oncol Rep, 32:1108–1116.
32.
TaiTW, SuFC, ChenCY, JouIM and LinCF. (2014). Activation of p38 MAPK-regulated Bcl-xL signaling increases survival against zoledronic acid-induced apoptosis in osteoclast precursors. Bone, 67:166–174.
33.
LeeHJ, ChoHS, JunSY, LeeJJ, YoonJY, LeeJH, SongHH, ChoiSH, KimSY, et al. (2014). Tussilago farfara L. augments TRAIL-induced apoptosis through MKK7/JNK activation by inhibition of MKK7TIPRL in human hepatocellular carcinoma cells. Oncol Rep, 32:1117–1123.
34.
ChanLY, ChiuPY and LauTK. (2004). Embryotoxicity study of ginsenoside Rc and Re in in vitro rat whole embryo culture. Reprod Toxicol, 19:131–134.
35.
ParkEJ, ZhaoYZ, KimJ and SohnDH. (2006). A ginsenoside metabolite, 20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol, triggers apoptosis in activated rat hepatic stellate cells via caspase-3 activation. Planta Med, 72:1250–1253.
36.
HeNW, ZhaoY, GuoL, ShangJ and YangXB. (2012). Antioxidant, antiproliferative, and pro-apoptotic activities of a saponin extract derived from the roots of Panax notoginseng (Burk.) F.H. Chen. J Med Food, 15:350–359.
37.
OyadomariS and MoriM. (2004). Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ, 11:381–389.
38.
PorterAG and JanickeRU. (1999). Emerging roles of caspase-3 in apoptosis. Cell Death Differ, 6:99–104.
39.
LinJ, XuH, WuY, TangM, McEwenGD, LiuP, HansenDR, GilbertsonTA and ZhouA. (2013). Investigation of free fatty acid associated recombinant membrane receptor protein expression in HEK293 cells using Raman spectroscopy, calcium imaging, and atomic force microscopy. Anal Chem, 85:1374–1381.
40.
OzdenerMH, SubramaniamS, SundaresanS, SeryO, HashimotoT, AsakawaY, BesnardP, AbumradNA and KhanNA. (2014). CD36- and GPR120-mediated Ca(2)(+) signaling in human taste bud cells mediates differential responses to fatty acids and is altered in obese mice. Gastroenterology, 146:995–1005.
41.
Abdoul-AzizeS, SelvakumarS, SadouH, BesnardP and KhanNA. (2014). Ca2+ signaling in taste bud cells and spontaneous preference for fat: unresolved roles of CD36 and GPR120. Biochimie, 96:8–13.
42.
HartmannC and TabinCJ. (2001). Wnt-14 plays a pivotal role in inducing synovial joint formation in the developing appendicular skeleton. Cell, 104:341–351.
43.
IchimuraA, HirasawaA, Poulain-GodefroyO, BonnefondA, HaraT, YengoL, KimuraI, LeloireA, LiuN, et al. (2012). Dysfunction of lipid sensor GPR120 leads to obesity in both mouse and human. Nature, 483:350–354.
44.
KorenN, Simsa-MazielS, ShaharR, SchwartzB and Monsonego-OrnanE. (2014). Exposure to omega-3 fatty acids at early age accelerate bone growth and improve bone quality. J Nutr Biochem, 25:623–633.
45.
TazoeH, OtomoY, KajiI, TanakaR, KarakiSI and KuwaharaA. (2008). Roles of short-chain fatty acids receptors, GPR41 and GPR43 on colonic functions. J Physiol Pharmacol, 59Suppl 2:251–262.
46.
WangJ, WuX, SimonaviciusN, TianH and LingL. (2006). Medium-chain fatty acids as ligands for orphan G protein-coupled receptor GPR84. J Biol Chem, 281:34457–34464.
47.
ItohY, KawamataY, HaradaM, KobayashiM, FujiiR, FukusumiS, OgiK, HosoyaM, TanakaY, et al. (2003). Free fatty acids regulate insulin secretion from pancreatic beta cells through GPR40. Nature, 422:173–176.
48.
HirasawaA, TsumayaK, AwajiT, KatsumaS, AdachiT, YamadaM, SugimotoY, MiyazakiS and TsujimotoG. (2005). Free fatty acids regulate gut incretin glucagon-like peptide-1 secretion through GPR120. Nat Med, 11:90–94.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
