Ethyl acetate fraction of Huogu formula inhibits adipogenic differentiation of bone marrow stromal cells via the BMP and Wnt signaling pathways

Abstract

Elevated adipogenesis of bone marrow stromal cells (BMSCs) is closely associated with non-traumatic osteonecrosis of femoral head (ONFH). Our previous studies have shown that Huogu (HG) formula was effective both in clinic experience and experimental ONFH. How HG impacts the differentiation of BMSCs and what is the underlying molecular mechanism remain largely unknown. Our results showed that ethyl acetate extract of HG (HGE) significantly decreased the adipocyte differentiation as determined by oil red staining, while slightly increased the ALP activity. Investigation of the molecular mechanism revealed that HGE could inhibit the mRNA and protein expression of peroxisome proliferators-activated receptor (PPAR)γ, lipoprotein lipase (LPL) and adipocyteprotein2 (AP2). Interestingly, the inhibition of adipogenic differentiation in BMSCs by HGE could be restored by DKK-1, an inhibitor of Wnts. However, Noggin (an inhibitor of BMPs) displayed an additive role with HGE in suppressing the expression of PPARγ, LPL, and AP2. Furthermore, the bone marrow fat formation, as well as the expression of Wnt3a and PPARγ, was effectively regulated by HGE in the steroid-induced ONFH rats. Our results demonstrated that HGE treatment significantly inhibited adipogenesis and slightly promoted osteogenesis of BMSCs through regulating the BMP and Wnt pathways. The findings shed lights on the molecular mechanism of HGE in the inhibition of adipogenesis and provide scientific rationale for its clinical application of HGE in the treatment of ONFH.

Cardioprotective effects of anisodamine against myocardial ischemia/reperfusion injury through the inhibition of oxidative stress, inflammation and apoptosis

“…During myocardial I/R injury, ROS production from the mitochondria and Nox is implicated in oxidative stress injury in cardiomyocytes (22). Nox is an enzyme also expressed in phagocytes; however, Nox in myocardial cells differs in its catalytic characteristics and biological functions: Cardiac muscle Nox is weakly active under physiological conditions, whereas ROS production is a critical function of phagocytosis; cardiac Nox and NADPH or nicotinamide adenine dinucleotide (NADH) are electron donors, whereas only NADPH is an electron donor in phagocytes; in phagocytes, Nox-generated ROS mainly participate in the host mechanisms of defense (23,24), whereas in cardiomyocytes ROS function as a second messenger during the regulation of cellular proliferation and differentiation (22). The results of the present study revealed that anisodamine significantly reversed the I/R-induced decrease in SOD levels and increase in MDA levels in serum samples isolated from rats following I/R.…”

“…Due to differences in their activation mechanisms, Nox2 needs to be induced in order to generate O 2-, whereas Nox4 can constitutively generate low levels of H 2 O 2 (26). Furthermore, Nox4 in cardiac muscle cells can catalyze O 2generation effectively, using NADH as a hydrogen donor (23). Due to the autonomous activity of Nox4, NADH is highly used in cardiac muscle cells (26).…”

Wnt5a/FZD4 Mediates the Mechanical Stretch-Induced Osteogenic Differentiation of Bone Mesenchymal Stem Cells

“…The importance of both canonical and noncanonical Wnt signalling in the osteogenesis of stem cells has been widely explored in many studies in vivo and in vitro [12,13]. Wnt5a signalling suppresses adipogenesis by suppressing the transcriptional activity of PPAR-γ and promotes the differentiation of BMSCs into osteoblasts [14,15].…”

Peripheral Blood Stem Cell Therapy Does Not Improve Outcomes of Femoral Head Osteonecrosis With Cap‐Shaped Separated Cartilage Defect

“…Recent studies demonstrated that ischemia, caused by interruption of the blood supply to the femoral head, is a primary cause of ONFH . In addition to osteocyte apoptosis, bone resorption, and adipocyte accumulation are also components of the progression of this disease …”