Androgen receptor over expression drives lipid accumulation in human hepatocytes

Nikolaos Nikolaou; Maryam Nasiri; Laura Gathercole; Silvia Parajes Castro; Nils Krone; George Valsamakis; George Mastorakos; Jeremy Tomlinson

doi:10.1530/endoabs.34.P363

Non-alcoholic fatty liver disease NAFLD has been associated with androgen deficiency, yet in the majority of patients with non-alcoholic steatohepatitis NASH, androgens levels are normal. In contrast, women with polycystic ovarian syndrome PCOS, which is characterised by androgen excess, have evidence of increased liver fat. Our hypothesis is that androgen exposure to the liver may be crucial in the amount of lipid that can accumulate in hepatocytes. C3A human hepatoma cells were cultured and treated with testosterone testo (5 and 50 nM) or dihydrotestosterone DHT (1 and 10 nM) for 24 h. Lipid accumulation was measured by C¹⁴ acetate incorporation and gene expression by real-time PCR. Cells were transfected with an androgen receptor AR construct (pcDNA3.1+AR) or vector as a control. FASN, ACC1, ACC2, and CPT1 mRNA expression was significantly increased after dose dependent treatment with testosterone and DHT (Table 1) and this was further augmented after AR over expression (FASN: ctrl 13.90±1.99 vs AR 66.78±6.22, ACC1: ctrl 1.06±0.26 vs AR 3.52±0.29, ACC2: ctrl 0.49±0.11 vs AR 1.02±0.09, and CPT1: ctrl 1.78±0.25 vs AR 4.33±0.22, P<0.05). Both testosterone and DHT increased de novo lipogenesis and this was also increased following AR over expression (7001.8±258.58 ctrl vs 8.747.76±433.39 Testo, 8.970.03±330.17 DHT, and AR 14193.2±755.17 AR, P<0.05). In conclusion, these data highlight that enhanced androgen action is able to drive lipid accumulation in human hepatocytes and this may be crucial in understanding the association between PCOS and NAFLD.


Gene of interest	Control (mean AU±S.E.M)	5 nM Testosterone (mean AU±S.E.M)	50 nM Testosterone (mean AU±S.E.M)	P value (5 nM/50 nM)
FASN	13.90±1.99	41.97±5.80*	60.36±3.34*	0.05/0.008
ACACA (ACC1)	1.05±0.26	2.03±0.56	3.06±0.41*	0.246/0.016
ACACB (ACC2)	0.50±0.11	0.78±0.14	0.96±0.15	0.955/0.15
CPT1	1.78±0.25	1.92±0.14	4.19±0.89*	0.89/0.039
Gene of interest	Control (mean AU±S.E.M)	1 nM of DHT (mean AU±S.E.M)	10 nM DHT (mean AU±S.E.M)	P value (1 nM/10 nM)
FASN	13.90±1.99	60.63±11.02**	81.97±19.46**	0.0008/0.0002
ACACA (ACC1)	1.06±0.26	3.72±0.96	4.45±1.09*	0.095/0.049
ACACB (ACC2)	0.49±0.11	1.01±0.18	1.31±0.16*	0.13/0.03
CPT1	1.78±0.25	3.25±0.12*	4.71±0.46**	0.019/0.002

Endocrine Abstracts

P363