Table 1.
Primers used in RT-qPCR analysis
| Gene | Primer sequence (5′-3′) | Product size (base pairs) | GenBank accession No. |
|---|---|---|---|
| GAPDH | F: CCTGGAGAAACCTGCAAAATA R: AACCTGGTCCTCAGTGTAGCC | 100 | NM_001206359.1 |
| TLR4 | F: GACGAAGACTGGGTGAGGAATGAAC R: CCTGGATGATGTTAGCAGCGATGG | 124 | NM_001113039.2 |
| MyD88 | F: CGTCTGGTCCATTGCTAGAACTC R: TTCTGATGGGCACCTGGAGAGAG | 141 | NM_001099923.1 |
| NF-κB | F: CTGAGGCTATAACTCGCTTGGTGAC R: CATGTCCGCAATGGAGGAGAAGTC | 131 | NM_001114281.1 |
| TNF-α | F: TCTATTTTGGGATCATTGCCC R: CCAGCCCCTCATTCTCTTTCT | 127 | NM_214022.1 |
| IL-2 | F: ACTTTCCAGGATGCTCAC R: ACTTCCTCCAGAGGTTTG | 102 | DQ852342.1 |
| IL-6 | F: ATAAGGGAAATGTCGAGGCTGTGC R: GGGTGGTGGCTTTGTCTGGATTC | 93 | NM_001252429.1 |
| IL-10 | F: GCATCCACTTCCCAACCA R: GCAACAAGTCGCCCATCT | 108 | NM_214041.1 |
| JAK1 | F: GACCGTCACCTGCTTTGAGA R: ACGAAGCTGATGTTGTCCGT | 104 | AB00601 |
| STAT3 | F: AGCCTCTCCGCAGAGTTCAA R: GCCCCCGTTCCCACAT | 60 | NM_001044580.1 |
| iNOS | F: CAGCGGGATGACTTTCCAA R: TTGCAAGCAAGATCCCCTTT | 60 | NM_001143690.1 |
| Keap1 | F: ACGACGTGGAGACAGAAACGT R: GCTTCGCCGATGCTTCA | 56 | NM_001114671.1 |
| Nrf2 | F: GCCCCTGGAAGCGTTAAAC R: GGACTGTATCCCCAGAAGGTTGT | 67 | XM_003133500.5 |
1 GAPDH – glyceraldehyde 3-phosphate dehydrogenase; TLR4 – toll-like receptor 4; MyD88 – myeloid differentiation factor 88; NFκB – nuclear factor kappa B; TNF-α – tumour necrosis factor α; IL – interleukin; JAK1 – janus kinase 1; STAT3 – signal transducer and activator of transcription 3; iNOS – inducible nitric oxide synthase; Keap1 – kelch-like ECH-associated protein 1; Nrf2 – nuclear factor erythroid 2-related factor 2; F – forward; R – reverse
Fig. 1.
The nitric oxide (NO) secretion in the thymus of weaned piglets. CON – control group; LPS – lipopolysaccharide group; * – significant increase (P-value < 0.05); ** – highly significant increase (P-value < 0.01)
Table 2.
The antioxidant parameters in the thymus of weaned piglets
| Group | Day 1 | Day 5 | Day 9 | Day 13 | |
|---|---|---|---|---|---|
| T-AOC (mmol/g protein) | CON | 0.1462 ± 0.00640 | 0.1077 ± 0.02364 | 0.0755 ± 0.01391 | 0.1027 ± 0.00941 |
| LPS | 0.1527 ± 0.01974 | 0.1189 ± 0.00409 | 0.0787 ± 0.00503 | 0.0960 ± 0.00266 | |
| CAT (U/mg protein) | CON | 1.3091 ± 0.12010 | 1.0701 ± 0.09588 | 1.1309 ± 0.04572 | 0.5535 ± 0.08402 |
| LPS | 2.2744 ± 0.16687** | 1.3503 ± 0.14927* | 1.0992 ± 0.16026 | 0.6420 ± 0.09374 | |
| SOD (U/mg protein) | CON | 15.4502 ± 1.37575 | 14.0568 ± 0.18257 | 9.0334 ± 0.31115 | 9.7733 ± 0.53686 |
| LPS | 14.4014 ± 1.02473 | 14.3906 ± 0.49276 | 9.8023 ± 0.31115* | 9.3018 ± 0.20602 | |
| GSH-Px (U/mg protein) | CON | 15.5310 ± 1.45508 | 13.7160 ± 0.42498 | 13.5570 ± 0.60307 | 16.8665 ± 1.07325 |
| LPS | 22.0999 ± 1.2071* | 18.0544 ± 1.9736* | 17.5424 ± 2.1281* | 21.4042 ± 1.58232** | |
| MDA (nmol/mg protein) | CON | 0.3856 ± 0.02354 | 0.3444 ± 0.11768 | 0.2435 ± 0.08353 | 0.2268 ± 0.00967 |
| LPS | 0.3634 ± 0.04299 | 0.3772 ± 0.07952 | 0.1684 ± 0.4000 | 0.2365 ± 0.04973 |
Fig. 2.
The mRNA expression of thymic toll-like receptor 4 (TLR4) pathway, related inflammatory factors and inducible nitric oxide synthase (iNOS). IL – interleukin; MyD88 – myeloid differentiation factor 88; NFκB – nuclear factor kappa B; TNF-α – tumour necrosis factor α; CON – control group; LPS – lipopolysaccharide group; * – significant increase (P-value < 0.05); ** – highly significant increase (P-value < 0.01); # – significant decrease (P-value < 0.05); ## – highly significant decrease (P-value < 0.01)
Fig. 3.
The mRNA expression of thymic JAK-STAT pathway and Keap1/Nrf2 pathway. A – janus kinase 1 (JAK1); B – kelch-like ECH-associated protein 1 (Keap1); C –signal transducer and activator of transcription 3 (STAT3); D – nuclear factor erythroid 2-related factor 2 (Nrf2). * – significant increase (P-value < 0.05); ** – highly significant increase (P-value < 0.01); # – significant decrease (P-value < 0.05); ## – highly significant decrease (P-value < 0.01)