Introduction
Target | Gene | Function | Product ref. |
---|---|---|---|
Fatty acid synthase |
Fasn
| Catalyses the formation of long-chain fatty acids in fatty acid synthesis | QT00149240 |
Acetyl coA carboxylase |
Acaca
| Essential role in regulating fatty acid synthesis | QT01554441 |
Stearoyl-CoA desaturase 1 |
Scd1
| Catalyses a rate-limiting step in the synthesis of unsaturated fatty acids. Key enzyme in fatty acid metabolism. | QT00291151 |
Lipoprotein lipase |
Lpl
| Hydrolysis of triglycerides into free fatty acids | QT01750469 |
Hormone sensitive lipase |
Lipe
| Hydrolyses stored triglycerides to free fatty acids | QT00169057 |
3-hydroxy-3-methylglutaryl-CoA reductase |
Hmgcr
| Rate-controlling enzyme of the mevalonate pathway that produces cholesterol | QT01037848 |
Sterol regulatory element-binding protein 1 |
Srebf1
| Cholesterol biosynthesis and uptake, and fatty acid biosynthesis | QT00167055 |
Sterol regulatory element-binding protein 2 |
Srebf2
| Cholesterol biosynthesis and uptake, and fatty acid biosynthesis | QT01045870 |
Apolipoprotein E |
Apoe
| Lipoprotein metabolism and transport. | QT01043889 |
ATP-binding cassette transporter A1 |
Abca1
| Major regulator of cellular cholesterol efflux and phospholipid homoeostasis | QT00165690 |
ATP-binding cassette transporter G5 |
Abcg5
| Cellular cholesterol efflux, promote biliary excretion of sterols. | QT00157752 |
Insulin receptor substrate 1 |
Irs1
| Transmitting signals from the insulin and insulin-like growth factor-1 (IGF-1) receptors to intracellular pathways in insulin signalling | QT00251657 |
Hexokinase 2 |
Hk2
| Phosphorylates glucose to glucose 6-phosphate in the glycolytic pathway | QT00155582 |
Hexokinase 4 (Glucokinase) |
Gck
| Phosphorylates glucose to glucose 6-phosphate in the glycolytic pathway | QT00140007 |
Phosphofructokinase |
Pfk
| Converts fructose-6-phosphate to fructose-1,6-bisphosphate, one of the most important regulatory enzymes of glycolysis | QT00159754 |
Carbohydrate-responsive element-binding protein |
Chrebp
| Activates of several regulatory enzymes of glycolysis and lipogenesis | QT00125335 |
Glucose transporter 4 |
Glut4
| Cellular glucose transport | QT01044946 |
Glucose-6-phosphate dehydrogenase |
G6pdx
| Enzyme in the pentose phosphate pathway, often for tissues actively engaged in biosynthesis of fatty acids | QT01748957 |
Glycogen synthase |
Gys
| Converts glucose to glycogen for storage, regulating glycogen/glucose levels | QT00162099 |
Liver X receptor alpha |
Nr1h3
| Nuclear receptor transcription factor regulating cholesterol, fatty acid, and glucose homoeostasis | QT00113729 |
Peroxisome proliferator-activated receptor alpha |
Ppara
| Transcription factor and major regulator of lipid metabolism | QT00137984 |
Peroxisome proliferator-activated receptor gamma |
Pparg
| Regulates fatty acid storage and glucose metabolism | QT00100296 |
Beta 2 microglobulin |
B2m
| Reference gene | QT01149547 |
Glyceraldehyde 3-phosphate dehydrogenase |
Gapdh
| Reference gene | QT01658692 |
Materials and methods
Animals
Experimental design and tissue collection
Measurement of total testosterone and 17β-estradiol
Quantitative analysis of mRNA
Western immunoblotting
Antibody | Concentration | Diluent | Supplier |
---|---|---|---|
FASN | 1:500 | 2.5 % milk bsa in tbs | CST |
ACACA | 1:500 | 2.5 % milk bsa in tbs | CST |
ABCA1 | 1:250 | 2.5 % milk bsa in tbs | abcam |
APOE | 1:250 | 5 % milk in tbs | abcam |
GCK | 1:500 | 5 % milk in tbs | abcam |
PFK | 1:250 | 0.01 % milk bsa in tbs | Proteintech |
GLUT4 | 1:500 | 2.5 % milk bsa in tbs | CST |
HK2 | 1:500 | 2.5 % milk bsa in tbs | CST |
LXR | 1:500 | 1 % milk bsa in tbs | abcam |
G6PD | 1:500 | 0.01 % milk bsa in tbs | Sigma |
GAPDH | 1:5000 | 2.5 % milk bsa in tbs | abcam |
Calnexin | 1:1000 | 5 % milk in tbs | CST |
Statistical analysis
Results
Carbohydrate metabolism
Fat Metabolism | |||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Muscle | Liver | Subcutaneous Fat | Visceral Fat | ||||||||||
Gene | Symbol | XY-P | Tfm-P | Tfm-S100 | XY-P | Tfm-P | Tfm-S100 | XY-P | Tfm-P | Tfm-S100 | XY-P | Tfm-P | Tfm-S100 |
Acetyl CoA carboxylase alpha | ACACA | 1.28 ± 0.27 | 1.65 ± 0.46 | 0.77 ± 0.14 | 1.09 ± 0.13 |
2.49 ± 0.64*
| 1.30 ± 0.29 | 1.22 ± 0.28 | 1.15 ± 0.36 | 1.89 ± 0.62 | 1.07 ± 0.16 | 1.02 ± 0.27 | 0.95 ± 0.35 |
Fatty acid synthase | FASN | 1.52 ± 0.43 | 1.48 ± 0.56 | 0.56 ± 0.14 | 1.15 ± 0.17 |
11.42 ± 4.93*
| 2.99 ± 0.88 | 1.20 ± 0.28 | 2.89 ± 1.49 | 4.76 ± 1.90 | 1.12 ± 0.23 | 1.35 ± 0.36 | 0.87 ± 0.30 |
Stearoyl-CoA desaturase-1 | SCD1 | 1.06 ± 0.11 | 1.30 ± 0.24 | 1.37 ± 0.29 | 1.04 ± 0.14 | 2.45 ± 1.15 | 1.38 ± 0.15 | 1.2 ± 0.29 | 3.05 ± 1.37 | 0.70 ± 0.15 | 1.10 ± 0.23 |
4.99 ± 1.64*
|
0.94 ± 0.15
†
|
Lipoprotein lipase | LPL | 1.24 ± 0.31 | 0.90 ± 0.30 | 0.83 ± 0.20 | 1.07 ± 0.13 | 1.26 ± 0.45 | 0.90 ± 0.26 | 1.03 ± 0.08 |
0.70 ± 0.09*
| 0.81 ± 0.22 | 1.08 ± 0.13 | 0.99 ± 0.20 |
2.18 ± 0.36
†
|
Hormone sensitive lipase | Lipe | 1.20 ± 0.46 | 1.25 ± 0.24 | 1.18 ± 0.49 | 1.03 ± 0.08 | 1.24 ± 0.16 | 0.85 ± 0.10 | 1.17 ± 0.17 | 1.49 ± 0.29 | 1.14 ± 0.20 | 1.05 ± 0.14 | 1.10 ± 0.27 | 0.82 ± 0.21 |
Cholesterol Homeostasis | |||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Muscle | Liver | Subcutaneous Fat | Visceral Fat | ||||||||||
Gene | Symbol | XY-P | Tfm-P | Tfm-S100 | XY-P | Tfm-P | Tfm-S100 | XY-P | Tfm-P | Tfm-S100 | XY-P | Tfm-P | Tfm-S100 |
3-hydroxy-3-methyl-glutaryl-CoA reductase | HMGCoAr | _ | _ | _ | 1.06 ± 0.11 | 3.19 ± 2.09 | 1.15 ± 0.18 | 1.12 ± 0.19 | 0.88 ± 0.16 | 1.33 ± 0.37 | _ | _ | _ |
Sterol Regulatory Element-Binding Protein | SREBPf1 | 1.29 ± 0.26 | 0.74 ± 0.21 | 1.47 ± 0.39 | 1.13 ± 0.16 | 1.01 ± 0.19 | 1.32 ± 0.29 | 1.07 ± 0.17 |
0.36 ± 0.09**
|
0.86 ± 0.16
†
| 0.81 ± 0.13 | 0.59 ± 0.25 | 0.78 ± 0.13 |
Sterol Regulatory Element-Binding Protein | SREBPf2 | 1.17 ± 0.21 | 0.80 ± 0.28 | 1.99 ± 0.62 | 1.08 ± 0.13 | 1.44 ± 0.22 | 1.17 ± 0.16 | 1.07 ± 0.17 |
0.42 ± 0.08**
|
1.12 ± 0.33
c
| 0.80 ± 0.14 | 0.56 ± 0.21 | 0.79 ± 0.13 |
Apolipoprotein E | ApoE | _ | _ | _ | 1.07 ± 0.05 |
0.77 ± 0.09**
|
1.14 ± 0.11
†
| 1.02 ± 0.08 |
0.59 ± 0.13*
|
1.14 ± 0.16
†
| 1.03 ± 0.10 | 0.75 ± 0.16 | 0.87 ± 0.06 |
ATP-binding cassette transporter A1 | ABCA1 | _ | _ | _ | 1.05 ± 0.04 |
0.71 ± 0.09**
|
1.10 ± 0.13
†
| 1.07 ± 0.14 | 0.77 ± 0.12 | 0.63 ± 0.11 | 1.14 ± 0.24 | 0.98 ± 0.20 | 1.19 ± 0.32 |
ATP-binding cassette transporter G5 | ABCG5 | _ | _ | _ | 1.01 ± 0.06 | 0.81 ± 0.15 | 1.01 ± 0.08 | _ | _ | _ | _ | _ | _ |
Carbohydrate Metabolism | |||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Muscle | Liver | Subcutaneous Fat | Visceral Fat | ||||||||||
Gene | Symbol | XY-P | Tfm-P | Tfm-S100 | XY-P | Tfm-P | Tfm-S100 | XY-P | Tfm-P | Tfm-S100 | XY-P | Tfm-P | Tfm-S100 |
Insulin receptor substrate 1 | IRS1 | 1.34 ± 0.32 | 1.37 ± 0.59 | 1.85 ± 0.94 | 1.08 ± 0.17 | 1.74 ± 0.54 | 1.13 ± 0.29 | _ | _ | _ | _ | _ | _ |
Hexokinase 2 | HK2 | 1.18 ± 0.19 |
0.50 ± 0.16*
| 0.54 ± 0.10 | _ | _ | _ | 1.32 ± 0.36 |
0.24 ± 0.05**
|
0.61 ± 0.19
†
| 0.97 ± 0.23 | 1.00 ± 0.21 | 1.57 ± 0.54 |
Hexokinase 4 (Glucokinase) | GCK | _ | _ | _ | 1.07 ± 0.10 |
0.47 ± 0.14**
|
0.97 ± 0.13
†
| _ | _ | _ | _ | _ | _ |
Phosphofructokinase | PFK | 1.28 ± 0.23 |
0.64 ± 0.16*
| 0.62 ± 0.10 | 1.19 ± 0.11 |
0.79 ± 0.15*
| 0.77 ± 0.06 | 1.76 ± 0.68 |
0.16 ± 0.05*
| 0.54 ± 0.31 | _ | _ | _ |
mitogen-activated protein kinase kinase 1 | MAP2K1 | 1.22 ± 0.22 |
0.65 ± 0.19
a
| 1.09 ± 0.21 | 1.09 ± 0.14 | 1.01 ± 0.14 |
1.62 ± 0.21
†
| _ | _ | _ | _ | _ | _ |
Carbohydrate regulatory element binding protein | ChREBP | 1.25 ± 0.24 | 2.16 ± 1.02 | 1.16 ± 0.22 | 1.05 ± 0.12 | 1.17 ± 0.13 | 1.11 ± 0.13 | 1.00 ± 0.19 | 1.76 ± 0.47 | 1.53 ± 0.38 | 1.33 ± 0.38 | 1.96 ± 0.7 | 1.76 ± 0.37 |
Glucose transporter 4 | GLUT4 | 1.20 ± 0.19 |
0.59 ± 0.14*
| 0.71 ± 0.13 | _ | _ | _ | 1.31 ± 0.32 |
0.37 ± 0.12*
| 0.73 ± 0.29 | 1.03 ± 0.10 | 1.24 ± 0.26 | 0.83 ± 0.16 |
glucose-6-phosphate 1-dehydrogenase X | G6PDx | 1.13 ± 0.19 | 1.72 ± 0.38 | 0.92 ± 0.12 | 1.03 ± 0.07 |
1.99 ± 0.20***
|
1.45 ± 0.18
b
| 1.06 ± 0.23 | 2.67 ± 1.36 | 1.09 ± 0.16 | 1.57 ± 0.59 | 2.53 ± 0.87 | 1.69 ± 0.4 |
Glycogen synthase | Gys1 | 1.14 ± 0.15 | 1.01 ± 0.31 | 1.82 ± 0.72 | 1.18 ± 0.18 | 2.13 ± 0.65 | 1.56 ± 0.22 | _ | _ | _ | _ | _ | _ |
Master Regulators | |||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Muscle | Liver | Subcutaneous Fat | Visceral Fat | ||||||||||
Gene | Symbol | XY-P | Tfm-P | Tfm-S100 | XY-P | Tfm-P | Tfm-S100 | XY-P | Tfm-P | Tfm-S100 | XY-P | Tfm-P | Tfm-S100 |
Liver X receptor | LXR | 1.35 ± 0.29 |
0.62 ± 0.12*
|
1.50 ± 0.27
††
| 1.05 ± 0.09 |
0.66 ± 0.08**
|
1.28 ± 0.24
†
| 1.07 ± 0.14 |
0.42 ± 0.13**
|
1.15 ± 0.28
†
| 1.27 ± 0.35 | 1.00 ± 0.30 | 1.44 ± 0.48 |
Peroxisome proliferator-activated receptor alpha | PPARa | 1.16 ± 0.20 | 1.65 ± 0.78 | 0.94 ± 0.21 | 1.02 ± 0.07 | 1.84 ± 0.55 | 0.93 ± 0.14 | 1.13 ± 0.23 | 0.41 ± 0.11 | 0.69 ± 0.22 | 1.09 ± 0.20 | 0.92 ± 0.32 | 0.98 ± 0.22 |
Peroxisome proliferator-activated receptor gamma | PPARg | 1.29 ± 0.31 | 1.29 ± 0.62 | 0.63 ± 0.17 | 1.09 ± 0.14 | 0.99 ± 0.19 | 1.29 ± 0.23 | 1.06 ± 0.14 | 0.67 ± 0.06 | 0.82 ± 0.12 | 1.05 ± 0.11 |
0.49 ± 0.08***
| 0.66 ± 0.10 |