EGCG Attenuates Renal Damage via Reversing Klotho Hypermethylation in Diabetic db/db Mice and HK-2 Cells
Detection of Klotho promoter methylation level in the kidney tissues of mice and HK-2 cells by pyrosequencing. (a) Significant changes of 14 CpG sites in the Klotho promoter of mice (chr5:151755140-151756066; mm9/mouse). (b) The methylation level of 6 CpG sites in each group of mice. Normal group: nontreated C57 mice. Control group: nontreated db/db mice. EGCG group: db/db mice treated with EGCG (50 mg/kg/d). (c) Representative pyrograms. The upper pyrogram is the normal group, the middle pyrogram is the control group, and the lower pyrogram is the EGCG group. Values are expressed as . vs. control. vs. nontreated db/db mice. vs. nontreated C57 mice. (d) Significant changes in 7 CpG sites in the Klotho promoter of HK-2 cells (chr13: 33589850-33591428; hg19/human). (e) The methylation level of the Klotho promoter in each group of HK-2 cells. NG: normal glucose (5.5 mM); HG: high glucose (60 mM); EGCG: HK-2 cells treated with 1 μM EGCG. (f) Representative pyrograms. The upper pyrogram is the NG group, the middle pyrogram is the HG group, and the lower pyrogram is the EGCG group. Values are expressed as . vs. the HG group.