Hyperglycemia induces elevated expression of thyroid hormone binding protein in vivo in kidney and heart and in vitro in mesangial cells.
During a search for glucose-regulated abundant mRNAs in the diabetic rat kidney, we cloned thyroid hormone binding protein (THBP), also known as mu-crystallin or CRYM. The aim of this study was to investigate the effect of hyperglycemia/high glucose on the expression of THBP. THBP mRNA copy numbers were determined in kidneys and hearts of diabetic GK rats vs normoglycemic Wistar rats, and in human mesangial cells (HMCs) exposed to high glucose using real-time qPCR, and THBP protein levels were measured by Western blotting and immunofluorescence. Intracellular ROS was measured in THBP transfected cells using DCF fluorescence. Hyperglycemia significantly increased THBP mRNA in GK rat kidneys (326+/-50 vs 147+/-54, p<0.05), and hearts (1583+/-277 vs 191+/-63, p<0.05). Moreover, the levels of THBP mRNA increased with age and hyperglycemia in GK rat kidneys, whereas in normoglycemic Wistar rat kidneys there was a decline with age. High glucose significantly increased THBP mRNA (92+/-37 vs 18+/-4, p<0.005), and protein in HMCs. The expression of THBP as a fusion protein in transfected HMCs resulted in reduction of glucose-induced intracellular ROS. We have shown that THBP mRNA is increased in diabetic kidney and heart, is regulated by high glucose in renal cells, and appears to attenuate glucose-induced intracellular ROS. These data suggest that THBP may be involved in the cellular pathways activated in response to glucose. This is the first report linking hyperglycemia with THBP and suggests that the role of THBP in diabetic complications should be further investigated.
Al-Kafaji
Ghada
G
Diabetes Research Group, Division of Reproduction and Endocrinology, King's College London, UK.
Malik
Afshan N
AN
D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003459:Crystallins / Q000096:biosynthesis* / Q000235:genetics; D003928:Diabetic Nephropathies / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D006801:Humans; D006943:Hyperglycemia / Q000378:metabolism*; D007668:Kidney / Q000378:metabolism*; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D050527:Mesangial Cells / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D013963:Thyroid Hormones / Q000096:biosynthesis* / Q000235:genetics; D000090803:mu-Crystallins; D000097810:Thyroid Hormone-Binding Proteins
D002352:Carrier Proteins; D003459:Crystallins; D005947:Glucose; D008565:Membrane Proteins; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D013963:Thyroid Hormones; D000090803:mu-Crystallins; D000097810:Thyroid Hormone-Binding Proteins; C000716469:CRYM protein, human; C000716471:Crym protein, rat