Long-term exposure to abnormal glucose levels alters drug metabolism pathways and insulin sensitivity in primary human hepatocytes.
Hyperglycemia in type 2 diabetes mellitus has been linked to non-alcoholic fatty liver disease, which can progress to inflammation, fibrosis/cirrhosis, and hepatocellular carcinoma. Understanding how chronic hyperglycemia affects primary human hepatocytes (PHHs) can facilitate the development of therapeutics for these diseases. Conversely, elucidating the effects of hypoglycemia on PHHs may provide insights into how the liver adapts to fasting, adverse diabetes drug reactions, and cancer. In contrast to declining PHH monocultures, micropatterned co-cultures (MPCCs) of PHHs and 3T3-J2 murine embryonic fibroblasts maintain insulin-sensitive glucose metabolism for several weeks. Here, we exposed MPCCs to hypo-, normo- and hyperglycemic culture media for ~3 weeks. While albumin and urea secretion were not affected by glucose level, hypoglycemic MPCCs upregulated CYP3A4 enzyme activity as compared to other glycemic states. In contrast, hyperglycemic MPCCs displayed significant hepatic lipid accumulation in the presence of insulin, while also showing decreased sensitivity to insulin-mediated inhibition of glucose output relative to a normoglycemic control. In conclusion, we show for the first time that PHHs exposed to hypo- and hyperglycemia can remain highly functional, but display increased CYP3A4 activity and selective insulin resistance, respectively. In the future, MPCCs under glycemic states can aid in novel drug discovery and mechanistic investigations.
School of Biomedical Engineering, Colorado State University, Fort Collins, CO 80523, USA.|Department of Bioengineering, University of Illinois, Chicago, IL 60607, USA.
Matthew D
MD
Davidson
Department of Mechanical Engineering, Colorado State University, Fort Collins, CO 80523, USA.
Kimberly R
KR
Ballinger
School of Biomedical Engineering, Colorado State University, Fort Collins, CO 80523, USA.|Department of Bioengineering, University of Illinois, Chicago, IL 60607, USA.|Department of Mechanical Engineering, Colorado State University, Fort Collins, CO 80523, USA.
Salman R
SR
Khetani
D016475:3T3 Cells; D000418:Albumins / Q000032:analysis / Q000378:metabolism; D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D002460:Cell Line; D018920:Coculture Techniques; D019388:Cytochrome P-450 CYP1A2 / Q000378:metabolism; D065637:Cytochrome P-450 CYP2A6 / Q000378:metabolism; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis / Q000378:metabolism*; D003924:Diabetes Mellitus, Type 2 / Q000097:blood; D005947:Glucose / Q000378:metabolism*; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D006943:Hyperglycemia / Q000473:pathology*; D007003:Hypoglycemia / Q000473:pathology*; D007328:Insulin / Q000378:metabolism; D007333:Insulin Resistance / Q000502:physiology; D008055:Lipids / Q000032:analysis; D008657:Metabolic Clearance Rate / Q000502:physiology*; D051379:Mice; D065626:Non-alcoholic Fatty Liver Disease / Q000097:blood; D014508:Urea / Q000032:analysis
D000418:Albumins; D001786:Blood Glucose; D007328:Insulin; D008055:Lipids; D014508:Urea; C504573:CYP1A2 protein, human; C585710:CYP2A6 protein, human; D019388:Cytochrome P-450 CYP1A2; D065637:Cytochrome P-450 CYP2A6; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D005947:Glucose