Direct and Indirect Effects of Guggulsterone on the Induction of Beiging in Mature 3T3-L1 Adipocytes
Direct and Indirect effects of Guggulsterone on the Induction
of Beiging in Mature 3T3-L1A Adipocytes B
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t Control GS 6 µM GS 25 µM
1 2 2 2 2 n 2 e Colette N. Miller, Yusra Azhar, Ashish Parmar, Janaiya Samuels, Rangaiaht 1 1 0 Shashidharamurthy,* Srujana Rayalam
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Department of Foods and Nutrition, University of Georgia, Athens, GA Department of Pharmaceutical Sciences, School of a Pharmacy, Philadelphia College of Osteopathic Medicine- GA Campus, Suwanee, GA i
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Abstract Direct Effects of GS on 3T3-L1 AdipocytesM GS signaling 9 0 T B X 1 Phytochemicals have long demonstrated anti-obesity properties in C o n tro l G S 2 5 u M Phytochemicals may induce beiging through nuclear receptor activation. adipocytes. Their ability, however, to induce browning in white adipose GS induces mitochondrial biogenesisC D T u b lin tissue is only beginning to emerge. We have recently established that the white adipocyte cell line, 3T3-L1, is capable of beiging under beta- 5 0 0 2 0 0 Control Iso 10mM p = 0 .1 8 adrenergic conditions. Using this information, we sought to investigate if the B p = 0 .1 8 A 4 0 0
4 0 0 1 5 0 )
plant steroid guggulsterone (GS) can induce beiging in 3T3-L1s. 3T3-L1 ) *
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o o r preadipocytes were differentiated using established protocols r *
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supplemented with rosiglitazone and thyroid hormone. Direct effects of GS n 3 0 0
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were measured by treating mature 3T3-L1s for 24 hours. Indirect effects o
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n were measured by treating mature 3T3-L1s with conditioned media from GS 6.25μM
GS 25μM 2 0 0 o
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GS-treated RAW 264.7 macrophages. Direct treatment of 3T3-L1s with GS
T B X 1 % resulted in increased lipolysis, increased mitochondrial activity (11%), and ( increased peroxisome proliferator-activated receptor gamma coactivator 1- 1 000 0 C o n tro l G S 6 u M G S 2 5 u M C o n tro l G S 6 u M G S 2 5 u M alpha (PGC1α) levels by 250% more than control. 3T3-L1s also T u b liAn demonstrated an increase in uncoupling protein 1 (UCP1) expression by 1 2 0 0 GS induces adipocyte beiging 1 0 0 C o n tro l IS O G S 6 u M G S 2 5 u M
200% and beige marker, T-box protein 1 (TBX1) expression by 80% more * * * )
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than control. Furthermore, this was accompanied by increased levels of G r t
b 3 0 0 n 4 0 0 a GS treatment appears to i 6 0
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protein-coupled bile acid receptor (TGR5) and its downstream target v
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iodothyronine deiodinase 2 (DIO2). Treatment of RAW 264.7 macrophages C C o n tro l IS O G S 6 u M G S 2 5 u M in 3T3-L1s. 3 0 0
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with GS induced a 60% increase in catecholamine release into the media ) l
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compared to control. Using this conditioned media from macrophages, 3T3- U C P 1 r 0
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P o M M M M M n r L1 adipocytes increased the expression of DIO2 and UCP1 following 24 t u u u u u 2 0 0 o
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T C c 1 S
U G G G hours of incubation. Results from this study demonstrate that GS can S G T B X 1 G
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% ( potentially induce beiging in white adipose tissue through two distinct ( 1 0 0 mechanisms: (1) direct signaling through the TGR5-cAMP-DIO2 pathway T u b lin B * * * * * * * and (2) indirectly through stimulating catecholamine release in 1 2 0 Working Model 0 0 macrophages. Thus, it is reasonable to conclude that GS may improve the B 1 0 0 # A C o n tro l IS O G S 6 u M G S 2 5 u M C o n tro l IS O G S 6 u M G S 2 5 u M
1 2 0 )
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metabolic capacity of adipose tissue thereby counteracting the effects of * i 8 0
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Indirect Effects1 of GS on 3T3-L1 Adipocytes Research Goals t r 1 0 0 t /m u u u u
n g 1 6 1 6 n X 0 o u S S S S C 1 o G G G G B l S o M M M M M + + c r u u u u u P • Determine if the anti-obesity phytochemical, guggulsterone, can T t S S L 0 n 1 3 6 .5 5 P P o 2 2 C S S S L L G G G 1 S A C o n tro l IS O G S 6 u M G SGS 2 5 u M protects against% 1 0 0 LPS-injury in RAWG 264.7 macrophages S promote mitochondrial biogenesis and beiging in 3T3-L1 adipocytes. ( G C C • Investigate if guggulsterone1 2 0 mediates mitochondrial biogenesis 3 0 0 * through direct or indirect signaling. B 5 0 0 1 0 0 1 2 0 * * * * * * *
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Direct4 0 Effects of GS on C
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2 0 N 0 0 0 C o n tro l IS O G S 6 u M G S 2 5 u M l l 3T3-L1 Adipocytes o L M M M M o L M M M r u u r 0 t /m u u t /m u u u n g 1 6 1 6 n g 6 5 5 o u o 2 2 S S S S u S C o n tro l G S 6 u M G S 1 2 .5 u M G S 2 5 u M C 1 C G G G G 1 G S S Conclusions S + + S G G P L S S P + P P L S L L P B L GS appears to have 2 distinct effects on adipocyte beiging: GS decreases adipogenesis C GS induces catecholamine secretion in RAW 264.7 macrophages that can upregulate 1) Direct signaling in adipocytes leading to upregulation of thermogenic/ 2 0 0 5 0 0
1 2 0 n UCP1 in adipocytes beiging makers and enhanced mitochondrial biogenesis. This effect
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