ROS Production and ERK Activity Are Involved in the Effects of d-β-Hydroxybutyrate and Metformin in a Glucose Deficient Condition.
Hypoglycemia, a complication of insulin or sulfonylurea therapy in diabetic patients, leads to brain damage. Furthermore, glucose replenishment following hypoglycemic coma induces neuronal cell death. In this study, we investigated the molecular mechanism underlying glucose deficiency-induced cytotoxicity and the protective effect of d-β-hydroxybutyrate (D-BHB) using SH-SY5Y cells. The cytotoxic mechanism of metformin under glucose deficiency was also examined. Cell viability under 1 mM glucose (glucose deficiency) was significantly decreased which was accompanied by increased production of reactive oxygen species (ROS) and decreased phosphorylation of extracellular signal-regulated kinase (ERK) and glycogen synthase 3 (GSK3β). ROS inhibitor reversed the glucose deficiency-induced cytotoxicity and restored the reduced phosphorylation of ERK and GSK3β. While metformin did not alter cell viability in normal glucose media, it further increased cell death and ROS production under glucose deficiency. However, D-BHB reversed cytotoxicity, ROS production, and the decrease in phosphorylation of ERK and GSK3β induced by the glucose deficiency. ERK inhibitor reversed the D-BHB-induced increase in cell viability under glucose deficiency, whereas GSK3β inhibitor did not restore glucose deficiency-induced cytotoxicity. Finally, the protective effect of D-BHB against glucose deficiency was confirmed in primary neuronal cells. We demonstrate that glucose deficiency-induced cytotoxicity is mediated by ERK inhibition through ROS production, which is attenuated by D-BHB and intensified by metformin.
Institute of Pharmaceutical Research and Development, College of Pharmacy, Wonkwang University, Iksan 570-749, Korea. lakki38786@gmail.com.|Hanbang Body-Fluid Research Center, Wonkwang University, Iksan 570-749, Korea. lakki38786@gmail.com.
Santosh
S
Lamichhane
Institute of Pharmaceutical Research and Development, College of Pharmacy, Wonkwang University, Iksan 570-749, Korea. tonkingbastola@gmail.com.|Hanbang Body-Fluid Research Center, Wonkwang University, Iksan 570-749, Korea. tonkingbastola@gmail.com.
Tonking
T
Bastola
Institute of Pharmaceutical Research and Development, College of Pharmacy, Wonkwang University, Iksan 570-749, Korea. ume.ramesh@gmail.com.|Hanbang Body-Fluid Research Center, Wonkwang University, Iksan 570-749, Korea. ume.ramesh@gmail.com.
Ramesh
R
Pariyar
Institute of Pharmaceutical Research and Development, College of Pharmacy, Wonkwang University, Iksan 570-749, Korea. Chori0509@naver.com.|Hanbang Body-Fluid Research Center, Wonkwang University, Iksan 570-749, Korea. Chori0509@naver.com.
Eun-Sol
ES
Lee
Hanbang Body-Fluid Research Center, Wonkwang University, Iksan 570-749, Korea. host@wku.ac.kr.|College of Oriental Medicine, Wonkwang University, Iksan 570-749, Korea. host@wku.ac.kr.
Ho-Sub
HS
Lee
Department of Internal Medicine, Gachon University Gil Medical Center, Incheon 21565, Korea. drhormone@naver.com.
Dae Ho
DH
Lee
Institute of Pharmaceutical Research and Development, College of Pharmacy, Wonkwang University, Iksan 570-749, Korea. jwseo@wku.ac.kr.|Hanbang Body-Fluid Research Center, Wonkwang University, Iksan 570-749, Korea. jwseo@wku.ac.kr.
Jungwon
J
Seo
D020155:3-Hydroxybutyric Acid / Q000494:pharmacology*; D000818:Animals; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D005947:Glucose / Q000172:deficiency*; D000071679:Glycogen Synthase Kinase 3 beta / Q000378:metabolism; D006801:Humans; D008687:Metformin / Q000633:toxicity; D051379:Mice; D008813:Mice, Inbred ICR; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism
D018696:Neuroprotective Agents; D017382:Reactive Oxygen Species; D008687:Metformin; D000071679:Glycogen Synthase Kinase 3 beta; D048049:Extracellular Signal-Regulated MAP Kinases; D005947:Glucose; D020155:3-Hydroxybutyric Acid
">d-β-hydroxybutyrate; extracellular signal-regulated kinase; hypoglycemia; metformin; reactive oxygen species