High Fat Diets Sex-Specifically Affect the Renal Transcriptome and Program Obesity, Kidney Injury, and Hypertension in the Offspring
Abstract
:1. Introduction
2. Materials and Methods
2.1. Experimental Design
2.2. Biochemical Analysis
2.3. Histology and Morphometric Study
2.4. Detection of l-arginine, l-citrulline, ADMA, and SDMA by HPLC
2.5. Next-Generation Sequencing and Analysis
2.6. Quantitative Real-time Polymerase Chain Reaction (PCR)
2.7. Western Blot
2.8. Immunohistochemistry Staining for 8-OHdG
2.9. Statistical Analysis
3. Results
3.1. Morphological Features and Biochemistry
3.2. Blood Pressure and Renal Outcome
3.3. Renal Transcriptome
3.4. Oxidative Stress and Nitric Oxide Pathway
3.5. RAS and Sodium Transporters
3.6. Clock and Clock-Controlled Genes
4. Discussion
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Gene | Forward | Reverse |
---|---|---|
Collagen I | 5 aggcataaagggtcatcgtg 3 | 5 accgttgagtccatctttgc 3 |
α-SMA | 5 gaccctgaagtatccgatagaaca 3 | 5 cacgcgaagctcgttatagaag 3 |
Ren | 5 aacattaccagggcaactttcact 3 | 5 acccccttcatggtgatctg 3 |
Atp6ap2 | 5 gaggcagtgaccctcaacat 3 | 5 ccctcctcacacaacaaggt 3 |
Agt | 5 gcccaggtcgcgatgat 3 | 5 tgtacaagatgctgagtgaggcaa 3 |
Ace | 5 caccggcaaggtctgctt 3 | 5 cttggcatagtttcgtgaggaa 3 |
Ace2 | 5 acccttcttacatcagccctactg 3 | 5 tgtccaaaacctaccccacatat 3 |
Agtr1a | 5 gctgggcaacgagtttgtct 3 | 5 cagtccttcagctggatcttca 3 |
Agtr1b | 5 caatctggctgtggctgactt 3 | 5 tgcacatcacaggtccaaaga 3 |
Mas1 | 5 catctctcctctcggctttgtg 3 | 5 cctcatccggaagcaaagg 3 |
Clock | 5 ccactgtacaatacgatggtgatctc 3 | 5 tgcggcatactggatggaat3 |
Bmal1 | 5 attccagggggaaccaga 3 | 5 gaaggtgatgaccctcttatcct 3 |
Per1 | 5 gcttgtgtggactgtggtagca 3 | 5 gccccaatccatccagttgt 3 |
Per2 | 5 catctgccacctcagactca 3 | 5 ctggtgtgacttgtatcactgct 3 |
Per3 | 5 tggccacagcatcagtaca 3 | 5 tacactgctggcactgcttc 3 |
Cry1 | 5 atcgtgcgcatttcacatac 3 | 5 tccgccattgagttctatgat 3 |
Cry2 | 5 gggagcatcagcaacacag 3 | 5 gcttccagcttgcgtttg 3 |
Ck1e | 5 gcctctatcaacacccacct 3 | 5 ggagcccaggttgaagtaca 3 |
Nr1d1 | 5 ctactggctccctcacccagga 3 | 5 gacactcggctgctgtcttcca 3 |
Rn18s | 5 gccgcggtaattccagctcca 3 | 5 cccgcccgctcccaagatc 3 |
Groups | ND/ND | ND/HF | HF/ND | HF/HF | p Value | |||
---|---|---|---|---|---|---|---|---|
Number | M = 6; F = 6 | M = 6; F = 6 | M = 6; F = 6 | M = 7; F = 6 | Pre | Post | Pre × Post | |
Body weight (g) | Male | 641 ± 39 | 785 ± 39 * | 677 ± 21 | 813 ± 41*,$ | NS | 0.001 | NS |
Female | 372 ± 17 | 362 ± 19 | 355 ± 14 | 372 ± 10 | NS | NS | NS | |
Left kidney (LK) weight (g) | Male | 2.16 ± 0.14 | 2.12 ± 0.08 | 2.26 ± 0.08 | 2.08 ± 0.07 | NS | NS | NS |
Female | 1.28 ± 0.04 | 1.45 ± 0.1 | 1.37 ± 0.05 | 1.38 ± 0.01 | NS | NS | NS | |
LK weight/100 g BW | Male | 0.34 ± 0.02 | 0.27 ± 0.01 * | 0.33 ± 0.01 # | 0.26 ± 0.01*,$ | NS | <0.001 | NS |
Female | 0.35 ± 0.02 | 0.4 ± 0.02 | 0.39 ± 0.01 | 0.37 ± 0.01 | NS | NS | 0.021 | |
AST (U/L) | Male | 88 ± 11 | 308 ± 58 * | 82 ± 10 # | 145 ± 19 # | 0.019 | <0.001 | 0.028 |
Female | 73 ± 3 | 160 ± 24 | 83 ± 12 | 82 ± 8 | 0.026 | 0.007 | 0.006 | |
ALT (U/L) | Male | 27 ± 3 | 196 ± 52 * | 23 ± 2 # | 66 ± 17 # | 0.031 | 0.002 | 0.041 |
Female | 19 ± 2 | 67 ± 13 | 22 ± 3 | 30 ± 4 | 0.026 | 0.001 | 0.009 | |
Total cholesterol (mg/dL) | Male | 71 ± 8 | 82 ± 7 | 58 ± 5 | 65 ± 5 | 0.027 | NS | NS |
Female | 81 ± 9 | 95 ± 4 | 104 ± 7 | 95 ± 16 | NS | NS | NS | |
Triglyceride (mg/dL) | Male | 101 ± 26 | 61 ± 13 | 105 ± 9 | 87 ± 12 | NS | NS | NS |
Female | 97 ± 23 | 58 ± 9 | 120 ± 23 # | 60 ± 8 | NS | NS | NS | |
HDL (mg/dL) | Male | 43 ± 4 | 49 ± 6 | 34 ± 4 | 42 ± 4 | NS | NS | NS |
Female | 39 ± 4 | 52 ± 2 | 59 ± 4 | 58 ± 10 | NS | NS | NS | |
Glucose (mg/dL) | Male | 81 ± 2 | 91 ± 3 | 93 ± 4 | 81 ± 3 | NS | NS | NS |
Female | 75 ± 4 | 76 ± 1 | 73 ± 2 | 62 ± 3 # | NS | NS | NS | |
IPGTT (AUC, mg/dL·120 min) | Male | 22,071 ± 1354 | 23,923 ± 2345 | 23,498 ± 2286 | 25,922 ± 1973 | - | - | - |
Female | 26,420 ± 1406 | 31,389 ± 1773 * | 26,890 ± 1820 | 26,949 ± 2416 | - | - | - |
KEGG Pathway | Count | Gene Symbol | p-Value | Benjamini |
---|---|---|---|---|
Male | ||||
Protein digestion and absorption | 2 | Slc15a1, Slc6a19 | 5.6 × 10−2 | 5.6 × 10−2 |
Female | ||||
Oxidative phosphorylation | 5 | Atp5j2, Atp6v0d2, Ndufa5, Cox6c, Cox7c | 1.6 × 10−2 | 1.6 × 10−2 |
Protein digestion and absorption | 4 | Slc15a1, Slc6a19, Slc7a7, Slc7a8 | 2.2 × 10−2 | 2.2 × 10−2 |
Metabolic pathways | 16 | Dhcr24, Abat, Atp5j2, Atp6v0d2, C1qalt1c1, Mgat4c, Ndufa5, Alox15, Cyp24a1, Cox6c, Cox7c, Dse, Dqkq, Gatm, Hykk, Polr2k | 2.3 × 10−2 | 2.3 × 10−2 |
Ribosome | 5 | Mrpl33, Mrps18c, Rpl22l1, Rpl30, LOC100362027 | 2.9 × 10−2 | 2.9 × 10−2 |
Cardiac muscle contraction | 3 | Cacna2d2, Cox6c, Cox7c | 9.9 × 10−2 | 9.9 × 10−2 |
Groups | ND/ND | ND/HF | HF/ND | HF/HF | P Value | |||
---|---|---|---|---|---|---|---|---|
Number | M = 5; F = 6 | M = 6; F = 6 | M = 6; F = 6 | M = 7; F = 6 | Pre | Post | Pre × Post | |
l-Citrulline (μM) | Male | 42.4 ± 2.2 | 42.0 ± 2.0 | 41.6 ± 1.6 | 46.2 ± 1.9 | NS | NS | NS |
Female | 48.7 ± 4.1 | 61.7 ± 9.5 | 44.5 ± 4.8 | 62.9 ± 4.7 | NS | 0.019 | NS | |
l-Arginine (μM) | Male | 168.0 ± 15.9 | 46.1 ± 12.8 * | 172.0 ± 9.5 # | 101.8 ± 15.5 | 0.041 | <0.001 | NS |
Female | 152.4 ± 27.7 | 120.8 ± 23.4 | 179.0 ± 10.2 | 119.6 ± 5.2 | NS | 0.027 | NS | |
ADMA (μM) | Male | 1.02 ± 0.03 | 0.92 ± 0.03 | 1.23 ± 0.03 # | 1.03 ± 0.07 | 0.001 | 0.003 | NS |
Female | 1.45 ± 0.09 | 1.25 ± 0.07 | 1.38 ± 0.03 | 1.1 ± 0.04 *,$ | NS | 0.001 | NS | |
SDMA(μM) | Male | 0.43 ± 0.03 | 0.55 ± 0.02 | 0.67 ± 0.03 * | 0.53 ± 0.02 | 0.001 | NS | <0.001 |
Female | 0.72 ± 0.08 | 0.65 ± 0.03 | 0.68 ± 0.05 | 0.5 ± 0.03* | NS | 0.028 | NS | |
l-Arginine-to-ADMA ratio (μM/μM) | Male | 165.7 ± 15.1 | 49.7 ± 13.9 * | 138.6 ± 7.7 # | 101.7 ± 17.6* | NS | <0.001 | 0.011 |
Female | 107.2 ± 20.8 | 93.3 ± 16.0 | 129.1 ± 9.6 | 107.9 ± 3.0 | NS | NS | NS | |
NOx (NO2− + NO−) (μM) | Male | 218.6 ± 15.1 | 167.5 ± 5.1 | 195.7 ± 6.6 | 178 ± 7.4 | NS | 0.001 | NS |
Female | 172.8 ± 16.5 | 161.5 ± 17.6 | 176.6 ± 16.5 | 159.1 ± 24.5 | NS | NS | NS |
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Tain, Y.-L.; Lin, Y.-J.; Sheen, J.-M.; Yu, H.-R.; Tiao, M.-M.; Chen, C.-C.; Tsai, C.-C.; Huang, L.-T.; Hsu, C.-N. High Fat Diets Sex-Specifically Affect the Renal Transcriptome and Program Obesity, Kidney Injury, and Hypertension in the Offspring. Nutrients 2017, 9, 357. https://doi.org/10.3390/nu9040357
Tain Y-L, Lin Y-J, Sheen J-M, Yu H-R, Tiao M-M, Chen C-C, Tsai C-C, Huang L-T, Hsu C-N. High Fat Diets Sex-Specifically Affect the Renal Transcriptome and Program Obesity, Kidney Injury, and Hypertension in the Offspring. Nutrients. 2017; 9(4):357. https://doi.org/10.3390/nu9040357
Chicago/Turabian StyleTain, You-Lin, Yu-Ju Lin, Jiunn-Ming Sheen, Hong-Ren Yu, Mao-Meng Tiao, Chih-Cheng Chen, Ching-Chou Tsai, Li-Tung Huang, and Chien-Ning Hsu. 2017. "High Fat Diets Sex-Specifically Affect the Renal Transcriptome and Program Obesity, Kidney Injury, and Hypertension in the Offspring" Nutrients 9, no. 4: 357. https://doi.org/10.3390/nu9040357