Table 2.
Real-time PCR confirmation of array results for select genes.
| Accession Number |
Gene Symbol |
Fold Δ | Description and/or functions | References | |
|---|---|---|---|---|---|
| PCR | Array | ||||
| NM_012532 | Cp | 4.3 | 3.3* | Ceruloplasmin; plasma metalloprotein made in liver that that binds copper, increased expression in diabetes, expressed in smooth muscle cells, inhibits eNOS, impairs vessel relaxation | 9, 17, 19, 42 |
| NM_139082 | Bambi | 4.2 | 4.7 | BMP and activin membrane-bound inhibitor, homolog (Xenopus laevis); inhibits TGFβ signaling | 87, 117 |
| XM_214551 | (Cidea) | 3.5 | 2.6 | Similar to cell death-inducing DFFA-like effector A; induces apoptosis | 54 |
| NM_053394 | Klf5 | 2.7 | 3.3 | Kruppel-like factor 5; transcription factor modulates SMC phenotype | 51, 122 |
| NM_012598 | Lpl | 2.7 | 2.6 | Multifunctional enzyme, widely expressed including vascular smooth muscle cells, Lpl protein is detected on endothelium (no mRNA), overexpression impairs blood vessel relaxation | 16, 27, 57 |
| XM_238119 | (Sema4b) | 2.6 | 6.2 | Similar to semaphorin 4B; class 4 semaphorins have transmembrane domain | 35 |
| NM_012520 | Cat | 2.4 | 2.2 | Catalase; antioxidant with expression in response to oxidative stress | 78 |
| NM_053698 | Cited2 | 2.4 | 2.5 | Cbp/p300-interacting transactivator, with Glu/Asp-rich terminal domain, 2; transcriptional cofactor, represses HIF1α activity, negative regulator of MMP1 and 13 | 36, 116, 129 |
| NM_007643 | Cd36 | 2.3 | 2.2 | Class B scavenger receptor, possible role in development of eNOS-related vascular dysfunction, absence may help protect against atherosclerosis | 33, 64 |
| NM_031535 | Bcl2l1 | 2.1 | 2.3 | Bcl2-like 1, BCL-2 protein family; anti-apoptotic signaling | 120 |
| NM_134455 | Cx3cl1 | 1.8 | 3.6 | Chemokine (C-X3-C motif) ligand 1 (also termed Fractalkine); upregulation in diabetic nephropathy, aortic SMC expression | 18, 63 |
| NM_017107 | Ogt | 1.8 | 1.5 | O-linked N-acetylglucosamine (GlcNAc) transferase; GlcNAc onto proteins | 2 |
| NM_053402 | Wnt4 | 1.8 | 2.2 | Glycoprotein involved in genitourinary development, increased in renal fibrosis and wound healing, experimental overexpression disrupts normal vascular pattern formation in the testis | 56, 69, 111 |
| NM_053578 | Atp6k | 1.0 | 1.0 | Vacuolar proton-ATPase subunit M9.2; vacuolar proton pump. No change detected. | |
| NM_012553 | Ela2 | 1.0 | 4.5 | Elastase 2; enzyme expressed in rat arterial beds, generates angiotensin II | 104 |
| NM_031825 | Fbn1 | −1.4 | −2.1 | Involved with microfibril assembly, is found in elastic connective tissues | 62 |
| J04035 | Eln | −1.6 | −1.7 | Elastin, also tropoelastin; elasticity of tissues, substrate for Lox | 62 |
| AY007691 | (Cspg2) | −1.9 | −3.0* | Proteoglycan in vascular wall, expressed by SMC, upregulated in vascular injury | 79, 83 |
| NM_012656 | Sparc | −2.0 | −2.2* | Absence associated with changes in ECM assembly/composition and is reduced in vessels with Monckeberg’s sclerosis (medial calcification) | 12, 106 |
| NM_012881 | Spp1 | −2.2 | −11.8 | Multiple reported roles in vascular calcification, remodeling, and vessel structure | 81, 121 |
| NM_173125 | (Pdlim7) | −2.6 | −3.1 | PDZ and LIM domain 7 (also termed Enigma); LIM domains bind to protein kinases, whereas PDZ domain binds to actin filaments, upregulated by estrogen in cultured vascular SMCs | 48, 123 |
| NM_017061 | Lox | −2.6 | −2.7* | Cross-links elastin and collagen, secreted by smooth muscle, vessel remodeling | 85, 109 |
| NM_012618 | S100a4 | −3.1 | −1.9 | S100 calcium-binding protein A4; expression detected in aorta and SMC | 24 |
| NM_019282 | Cktsf1b1 | −7.9 | −5.8 | Cysteine knot superfamily 1, BMP antagonist 1 (also termed Gremlin 1); induced in diabetic nephropathy, BMP antagonist actions increase FGF and sonic hedgehog expression | 61, 70 |
Real-time PCR validation of various genes estimated to be differentially expressed based on the Resolver array analysis (p-value ≤ 0.01). Atp6k was selected as a gene whose expression was unchanged between groups based on array results. For PCR, relative changes in gene expression (fold Δ) were calculated using the 2−ΔΔCT method. Also, PCR fold Δ is relative to control cDNA while using Actb (Beta actin) expression as a reference gene. Array fold Δ in certain cases (*) is an average since the gene had multiple probe sets called significant. (Gene Symbol) = Rattus norvegicus transcript listed as “similar to” mouse and human gene, Homologene-NCBI. SMC = smooth muscle cell.