Effect of Mollugin on Apoptosis and Adipogenesis of 3T3-L1 Preadipocytes

Laboratory of Immunobiology, School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University, Daegu, Korea.
Phytotherapy Research (Impact Factor: 2.66). 01/2010; 25(5):724-31. DOI: 10.1002/ptr.3329
Source: PubMed


The effect of mollugin, isolated from the roots of Rubia cordifolia L., on cell viability, apoptosis and adipogenesis in 3T3-L1 preadipocytes was investigated. The inhibitory effect of mollugin (40-60 µM) on cell viability was more significant in differentiated adipocytes than in 3T3-L1 preadipocytes. In 3T3-L1 cells, the cytotoxicity of mollugin was accompanied by apoptotic events including mitochondrial membrane potential (Δψm) loss and activation of caspase-9, -3 and -7, leading to PARP degradation. Although the presence of 20 µM mollugin during induced adipocytic differentiation of 3T3-L1 cells for 6 days failed to affect the cell viability, it could almost completely abrogate the differentiation-associated morphology change and intracellular lipid accumulation. A similar level of inhibition was observed, when 20 µM mollugin was present during the early stage (D0-D2) of the differentiation period. In addition, the expression of C/EBPα, PPARγ1 and PPARγ2 was significantly down-regulated. The presence of 20 µM mollugin during either middle stage (D2-D4) or late stage (D4-D6) of the differentiation period, however, caused the inhibition to a lesser extent. These results indicated that mollugin at high concentrations (40-60 µM) exerted cytotoxicity via inducing apoptosis, whereas mollugin at a low concentration (20 µM) suppressed adipocytic differentiation without exerting cytotoxicity in 3T3-L1 preadipocytes.


Available from: Young Ho Kim, Dec 26, 2015
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    • "2A and 2B, although the presence of each fraction at a concentration of 25 μg/ml, except for the butanol fraction, during the differentiation period for 6 days did not influence significantly the cell viability , it could reduce the intracellular lipid accumulation to a level of 81.5% ~50.3% of the control. Under these conditions , the Oil Red O staining, which was performed to visualize intracellular lipid accumulation during the induced adipocytic differentiation of 3T3-L1 cells [5, 13], showed that the presence of the butanol fraction during the 6-day differentiation period caused not only almost complete prevention of intracellular lipid accumulation, but also retaining of fibroblast-like morphology of 3T3-L1 preadipocytes (Fig. 2C). These Oil Red O staining data confirmed the inhibitory effect of the butanol fraction on differentiation of 3T3-L1 preadipocytes into mature adipocytes. "
    Full-text · Article · May 2014
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    • "In addition, this plant has been used in traditional Chinese medicine for the treatment of arthritis, dysmenorrheal, hematorrhea, hemostasis, and psoriasis [11, 12]. Among the bioactive components from Rubia cordifolia, mollugin (C17H16O4; methyl 2,2-dimethyl-6-hydroxy-2H-naphtho[1,2-b]pyran-5-carboxylate) has been reported to have antitumor and anti-inflammatory activities, and neuroprotective and apoptotic effects [13–16]. A recent study demonstrated that mollugin induced apoptosis through endoplasmic reticulum stress-mediated activation of c-Jun N-terminal kinase (JNK) and the mitochondria-dependent caspase cascade, regulated by Bcl-xL in human Jurkat T cells [14]. "
    [Show abstract] [Hide abstract] ABSTRACT: Although previous studies have shown that mollugin, a bioactive phytochemical isolated from Rubia cordifolia L. (Rubiaceae), exhibits antitumor effects, its biological activity in oral cancer has not been reported. We thus investigated the effects and putative mechanism of apoptosis induced by mollugin in human oral squamous cell carcinoma cells (OSCCs). Results show that mollugin induces cell death in a dose-dependent manner in primary and metastatic OSCCs. Mollugin-induced cell death involved apoptosis, characterized by the appearance of nuclear shrinkage, flow cytometric analysis of sub-G1 phase arrest, and annexin V-FITC and propidium iodide staining. Western blot analysis and RT-PCR revealed that mollugin suppressed activation of NF- κ B and NF- κ B-dependent gene products involved in antiapoptosis (Bcl-2 and Bcl-xl), invasion (MMP-9 and ICAM-1), and angiogenesis (FGF-2 and VEGF). Furthermore, mollugin induced the activation of p38, ERK, and JNK and the expression of heme oxygenase-1 (HO-1) and nuclear factor E2-related factor 2 (Nrf2). Mollugin-induced growth inhibition and apoptosis of HO-1 were reversed by an HO-1 inhibitor and Nrf2 siRNA. Collectively, this is the first report to demonstrate the effectiveness of mollugin as a candidate for a chemotherapeutic agent in OSCCs via the upregulation of the HO-1 and Nrf2 pathways and the downregulation of NF- κ B.
    Full-text · Article · May 2013
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    • "It represents a pivotal signaling protein in the apoptotic cascade. Inhibition of Caspase 9 by a chemical inhibitor mollugin affects adipogenesis [10, 11]. It is alternatively spliced to caspase 9a (apoptotic) and caspase 9b (antiapoptosis) via inclusion of the cassette exons 3, 4, 5, and 6 in caspase 9a (Figure 2(c)). "
    [Show abstract] [Hide abstract] ABSTRACT: Obesity and its comorbidities affect millions of people. Here, we demonstrate that human preadipocytes are susceptible to programmed cell death (apoptosis) while mature adipocytes are resistant to apoptosis. The molecular mechanisms underlying the phenotype of apoptosis-resistant adipocytes are lesser known. To study the role of apoptosis and define molecular differences in the developmental process of adipogenesis, human preadipocytes were differentiated in vitro to mature adipocytes. Many genes in the apoptosis pathway are alternatively spliced. Our data demonstrates that during differentiation PKC δ , Bclx, and Caspase9 switch to their prosurvival splice variants along with an increase in Bcl2 expression when the cells terminally differentiate into mature adipocytes. Next we determined the expression pattern of these genes in obesity. Our data indicated high expression of PKC δ VIII in adipose tissue of obese patient in different depots. We demonstrate a shift in the in vitro expression of these splice variants in differentiating preadipocytes derived from obese patients along with a decrease in adipogenesis markers. Hence, the programmed splicing of antiapoptotic proteins is a pivotal switch in differentiation that commits adipocytes to a prosurvival pathway. The expression pattern of these genes is dysregulated in obesity and may contribute to adipose tissue dysfunction.
    Full-text · Article · Apr 2013
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