Chlorogenic Acid Prevents Osteoporosis by Shp2/PI3K/Akt Pathway in Ovariectomized Rats

Abstract

Cortex Eucommiae is used worldwide in traditional medicine, various constituents of Cortex Eucommiae, such as chlorogenic acid (CGA), has been reported to exert anti-osteoporosis activity in China, but the mechanism about their contribution to the overall activity is limited. The aims of this study were to determine whether chlorogenic acid can prevent estrogen deficiency-induced osteoporosis and to analyze the mechanism of CGA bioactivity. The effect of CGA on estrogen deficiency-induced osteoporosis was performed in vivo. Sixty female Sprague-Dawley rats were divided randomly among a sham-operated group and five ovariectomy (OVX) plus treatment subgroups: saline vehicle, 17α-ethinylestradiol (E2), or CGA at 9, 27, or 45 mg/kg/d. The rats’ femoral metaphyses were evaluated by micro-computed tomography (μCT). The mechanism of CGA bioactivity was investigated in vitro. Bone mesenchymal stem cells (BMSCs) were treated with CGA, with or without phosphoinositide 3-kinase (PI3K) inhibitor LY294002. BMSCs proliferation and osteoblast differentiation were assessed with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and alkaline phosphatase, with or without Shp2 interfering RNA (RNAi). The results display that CGA at 27 and 45 mg/kg/day inhibited the decrease of bone mineral density (BMD) that induced by OVX in femur (p< 0.01), significantly promoted the levels of bone turnover markers, and prevented bone volume fraction (BV/TV), connectivity density (CoonD), trabecular number (Tb.N), trabecular thickness (Tb.Th) (all p< 0.01) to decrease and prevented the trabecular separation (Tb.Sp), structure model index (SMI)(both p< 0.01) to increase. CGA at 1 or 10 μM enhanced BMSC proliferation in a dose-dependent manner. CGA at 0.1 to 10 μM increased phosphorylated Akt (p-Akt) and cyclin D1. These effects were reversed by LY294002. CGA at 1 or 10 μM increased BMSC differentiation to osteoblasts (p< 0.01), Shp2 RNAi suppressed CGA-induced osteoblast differentiation by decreasing Shp2, p-Akt, and cyclin D1. This study found that CGA improved the BMD and trabecular micro-architecture for the OVX-induced osteoporosis. Therefore, CGA might be an effective alternative treatment for postmenopausal osteoporosis. CGA promoted proliferation of osteoblast precursors and osteoblastic differentiation of BMSCs via the Shp2/PI3K/Akt/cyclin D1 pathway.

Identification of Steroidogenic Components Derived From Gardenia jasminoides Ellis Potentially Useful for Treating Postmenopausal Syndrome

“…Although there is little information about the anti-cancer activity of GA, GA had a potential inhibitory effect on tumor growth (Hsu et al, 1997 ). For the inhibition of bone loss, rutin inhibited osteopenia through suppressing bone resorption and enhancing osteoblastic activity in ovariectomized (Ovx) rats (Horcajada-Molteni et al, 2000 ); CGA prevented the decrease of mineralization and promoted the increase the mechanical properties and thus inhibited bone loss in Ovx animals (Folwarczna et al, 2015 ; Zhou et al, 2016 ); GA promoted osteogenesis by increasing the proliferation of osteoblasts and inhibited osteolysis by decreasing the proliferation of osteoclasts (Ha et al, 2003 ). For the promotion of neuron regeneration, rutin was reported to prevent spatial and emotional memory impairment (Qu et al, 2014 ; Ramalingayya et al, 2016 ); CGA supplementation could interfere in neurological degeneration (Heitman and Ingram, 2017 ); Though less evidence indicates a direct connection of GA with cognitive improving activities, GA is the main component of Tong Luo Jiu Nao, which was shown to prevent neuronal damage and improve learning and memory (Liu et al, 2011 , 2013 ).…”

Network-Based Method for Identifying Co-Regeneration Genes in Bone, Dentin, Nerve and Vessel Tissues

“…PTPN11 (ENSP00000340944), encoding a functional protein tyrosine phosphatase with two SH2 domains, has been confirmed to be widely expressed in multiple tissue subtypes [101,102]. Also known as SHP2, such gene has been confirmed to contribute to the regeneration of nerve tissues by regulating Shp2/PI3K/Akt Pathway [103,104]. As for its contribution to angiogenesis, various publications confirmed that PTPN11 contributes to the generation of vessel cells, confirming its role for regeneration of vessel [105].…”

Dietary Bioactives and Bone Health

“…This compound antiproliferative activity (useful in neoplastic diseases) may explain the suppressive effect on accumulation of fibroblast-like synoviocytes. One in vitro study showed that the underlying mechanism may be represented by the apoptosis induction via increased production of reactive oxygen species and activation of p38 mitogen-activated protein kinase (MAPK) [60].…”

“…boswelic acids (−) PGE1-S, cathepsin G, LOX-5, MMP-9, MMP-13, COX-2, NO, PGE1, TNF-α, IL-1, IL-2, IL-4, IL-6, IFN-γ (in vitro, AM) [17,30,31,41] (−) leukocyte infiltration in knee (AM) [43] Curcuma spp. curcuminoids (+) SOD, GSH, (−) MDA (HS) [58] (−) neutrophil infiltrate in knee, (AM), (−) IL-1β, TNFα, MCP-1, and MIP-1α (in vitro, AM) [57,59] β-elemene (+) p38 MAPK (in vitro) [60] Equisetum arvense kynurenic acid (−) synoviocyte proliferation (in vitro) [64,66] Harpagophytum procumbens iridoid glycosides (−) iNOS and COX-2 (in vitro) [76] Panax notoginseng saponins (−) TNF-alpha, IL- …”

“…Previous research has demonstrated that chlorogenic acids are bone-protective. For instance, Zhou et al [60] demonstrated that supplementing Ovx rats with chlorogenic acid led to improved BMD and microarchitecture, and an increased proliferation of osteoblast precursors and osteoblast differentiation, as well as increases in bone formation biomarkers. Despite this, Léotoing et al [40] demonstrated that the bone-protective effects of dried plum is not dependent on the content of chlorogenic acids.…”