Bioconversion ofCitrus unshiupeel extracts with cytolase suppresses adipogenic activity in 3T3-L1 cells

Abstract

BACKGROUND/OBJECTIVESCitrus flavonoids have a variety of physiological properties such as anti-oxidant, anti-inflammation, anti-cancer, and anti-obesity. We investigated whether bioconversion of Citrus unshiu with cytolase (CU-C) ameliorates the anti-adipogenic effects by modulation of adipocyte differentiation and lipid metabolism in 3T3-L1 cells.MATERIALS/METHODSGlycoside forms of Citrus unshiu (CU) were converted into aglycoside forms with cytolase treatment. Cell viability of CU and CU-C was measured at various concentrations in 3T3L-1 cells. The anti-adipogenic and lipolytic effects were examined using Oil red O staining and free glycerol assay, respectively. We performed real time-polymerase chain reaction and western immunoblotting assay to detect mRNA and protein expression of adipogenic transcription factors, respectively.RESULTSTreatment with cytolase decreased flavanone rutinoside forms (narirutin and hesperidin) and instead, increased flavanone aglycoside forms (naringenin and hesperetin). During adipocyte differentiation, 3T3-L1 cells were treated with CU or CU-C at a dose of 0.5 mg/ml. Adipocyte differentiation was inhibited in CU-C group, but not in CU group. CU-C markedly suppressed the insulin-induced protein expression of CCAAT/enhancer-binding protein α (C/EBPα) and peroxisome proliferator-activated receptor gamma (PPARγ) as well as the mRNA levels of CEBPα, PPARγ, and sterol regulatory element binding protein 1c (SREBP1c). Both CU and CU-C groups significantly increased the adipolytic activity with the higher release of free glycerol than those of control group in differentiated 3T3-L1 adipocytes. CU-C is particularly superior in suppression of adipogenesis, whereas CU-C has similar effect to CU on stimulation of lipolysis.CONCLUSIONSThese results suggest that bioconversion of Citrus unshiu peel extracts with cytolase enhances aglycoside flavonoids and improves the anti-adipogenic metabolism via both inhibition of key adipogenic transcription factors and induction of adipolytic activity.

Hesperetin inhibits lipid accumulation and ROS production during adipocyte differentiation in 3T3-L1 cells

“…HT is the aglycone form of hesperidin that is commonly present in citrus fruits. A few studies have reported that HT exhibits beneficial properties, including antioxidant, anti‐inflammation, and antiadipogenic activities (Bok et al, ; Lim et al, ; Yang et al, ; Ye, Chan, Chen, & Leung, ). Yang et al investigated the antioxidant and anti‐inflammatory effects of HT metabolites derived from HT‐administered rat serum.…”

“…The flavonoid contents of CU include naringin, narirutin, and hesperidin, while CU‐C contains naringenin and hesperetin. The authors showed that CU and CU‐C suppressed three adipogenic transcription factors, including PPARγ, C/EBPα, and Sterol regulatory element‐binding protein‐1c (SREBP‐1c) in adipocyte differentiation (Lim et al, ). However, the effects of HT on lipid accumulation and ROS production as well as its molecular action during adipogenesis remain unclear.…”

Antioxidant effect of Kimchi supplemented with Jeju citrus concentrate and its antiobesity effect on 3T3‐L1 adipocytes

“…The absorbance of NK and CK was 0.39 and 0.27, respectively. Similarly, bioconversion of Citrus unshiu with cytolase (CU‐C) showed significant inhibitory effect on lipid accumulation at 0.5mg/ml (Lim et al, ). These results indicated that citrus concentrate inhibited lipid accumulation in 3T3‐L1 adipocytes, suggesting its antiobesity effect.…”

Effects of dietary dehydrated lemon peel on some biochemical markers related to general metabolism, welfare and stress in gilthead seabream (Sparus aurataL.)

“…Such kind of natural immunostimulants are medicinal plants, among these C. limon , and citrus plants in general, have long been used to treat many diseases in traditional medicine such as obesity, diabetes, hypercholesterolaemia, high blood pressure, inflammation (Padilla‐Camberos et al, ; Park et al, ; Parra et al, ; Tort, ), brain disorders, cancer (Lim et al, ; Parra et al, ; Shetty et al, ; Tort, ), microbial infections (Ou et al, ; Park et al, ; Shetty et al, ), digestive disorders (Padilla‐Camberos et al, ; Park et al, ; Shetty et al, ), vitamin deficiencies (Shetty et al, ) and bronchial, asthmatic and degenerative diseases (Lim et al, ) among others. In general, the different parts of citrus plants have beneficial properties, for example immune‐enhancing (Padilla‐Camberos et al, ; Shetty et al, ) and anti‐toxic (Chowdhury et al, ), and present antioxidant (Cho et al, ; Gómez et al, ; Nouri & Shafaghatlonbar, ; Ou et al, ; Padilla‐Camberos et al, ; Wu et al, ), antibacterial (Ou et al, ; Ramadan et al, ; Shetty et al, ), anti‐diabetic (Menichini et al, ; Padilla‐Camberos et al, ; Wu et al, ), anti‐inflammatory (Ramadan et al, ), hepato‐protective (Chowdhury et al, ), neuro‐protective (Ramadan et al, ) and kidney‐protective (Sridharan et al, ) activities due to the phenolic compounds they contain, such as terpenes (Nakajima et al, ; Ou et al, ; Park, Lee, & Park, ; Park et al, ; Shetty et al, ) phenolic acids (Ou et al, ; Park et al, ; Shetty et al, ) and flavonoids (Gómez et al, ; Lee et al, ; Lim et al, ; Nakajima et al, ; Okuyama & Zasshi, ; Park et al, ; Shetty et al, ; Wu et al, ). For these important reasons it was decided to test the effects of this important fruit on gilthead seabream, as on...…”

“…Due to the negative impact of drugs and/or antibiotics on fish and human health, natural immunostimulants that enhance the immune system response are increasingly emerging as alternatives to prevent fish diseases (Bulfon et al, 2015; Van Hai, 2015;Harikrishnan et al, 2011;Mena et al, 2013;Reverter et al, 2014). Such kind of natural immunostimulants are medicinal plants, among these C. limon, and citrus plants in general, have long been used to treat many diseases in traditional medicine such as obesity, diabetes, hypercholesterolaemia, high blood pressure, inflammation (Padilla-Camberos et al, 2014;Park et al, 2016;Parra et al, 2015;Tort, 2011), brain disorders, cancer (Lim et al, 2015;Parra et al, 2015;Shetty et al, 2016;Tort, 2011), microbial infections (Ou et al, 2015;Park et al, 2016;Shetty et al, 2016) Furthermore, DLP was chosen because it is one of the main by-products of the lemon processing industry and we wanted to determine whether it could be incorporated in the fish diet without any previous treatment, acting as a nutraceutic additive, as realized for other farmed animals (Santos et al, 2014).…”