1.Effects of Sulforaphane on the Proliferation and Apoptosis of Human Renal Tubular Epithelial Cells Induced by High Glucose and Its Mechanism
Lei ZHOU ; Wang AI ; Xiaojuan CHEN ; Chunyue WEN
China Pharmacy 2021;32(24):3000-3007
OBJECTIVE:To study the effects of sulforaphane on the prolifera tion and apoptosis of human renal tubular epithelial cells HK- 2 induced by high glucose ,and to investigate its mechanism primarily. METHODS :HK-2 cells were divided into normal group ,high glucose group ,irbesartan group (positive control ,1 μmol/L),sulforaphane low ,medium and high concentration groups (10,20,40 μmol/L). The cells in normal group were cultured in DMEM medium for 96 hours. T he cells in other groups were cultured in high glucose DMEM medium (containing 40 mmol/L glucose )for 48 hours. After inducing cell injury,the cells were added with corresponding drugs for 48 hours. Survival rate and apoptotic rate of cells were detected. mRNA expression of cyclin D 1,caspase-3,Bcl-2 and Bax as well as protein expression of p-mTOR ,p-AMPK,p-Akt and p-PI 3K were also determined. In addition ,HK-2 cells were divided into normal group ,high glucose group ,sulforaphane high concentration group(40 μmol/L),acardicin group (AMPK agonist ,1 mmol/L),sulforaphane high concentration+compound C group (sulforaphane 40 μmol/L+AMPK inhibitor compound C 40 μmol/L),perifoxine group (Akt inhibitor ,19.95 μmol/L)、sulforaphane high concentration+SC 79 group(sulforaphane 40 μmol/L+Akt agonist SC79 4 μmol/L). After cultured with the same method , protein expression of p-mTOR ,p-AMPK,p-Akt and p-PI 3K were detected in HK- 2 cells. RESULTS :Compared with normal group,survival rate of HK- 2 cells,mRNA expression of cyclin D 1 and Bcl- 2 as well as protein expression of p-AMPK were decreased significantly in high glucose group (P<0.05);apoptotic rate ,mRNA expression of caspase- 3 and Bax ,protein expression of p-mTOR ,p-Akt and p-PI 3K in HK- 2 cells were increased significantly (P<0.05). Compared with high glucose group,above indexes of sulforaphane low ,medium and high concentration groups ,irbesartan group were all improved significantly (P<0.05);the improvement of above indexes in sulforaphane medium and high concentration groups were significantly better those of sulforaphane low concentration group (P<0.05). There was no significant difference in above indexes between sulforaphane high concentration group and irbesartan group (P>0.05). Compared with sulforaphane high concentration group,there were no significant difference in the protein expression of p-AMPK ,p-mTOR in acardicin group and p-mTOR ,p-Akt and p-PI 3K in perifoxine group (P>0.05);the protein expression of p-AMPK in sulforaphane high concentration+compound C group was decreased significantly (P<0.05),while the protein expression of p-mTOR was increased significantly (P<0.05);the protein expression of p-mTOR 、p-Akt、p-PI3K in sulforaphane high concentration+SC 79 group were increased significantly (P< 0.05). CONCLUSIONS :Sulforaphane can promote the proliferation of renal tubular epithelial cells and inhibit its apoptosis ;its mechanism may be associated with up-regulating the expression of p-AMPK and down-regulating the expression of p-mTOR ,p-Akt and p-PI 3K.