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萝卜硫素对高糖诱导人肾小管上皮细胞增殖和凋亡的影响及机制
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| 篇名： | 萝卜硫素对高糖诱导人肾小管上皮细胞增殖和凋亡的影响及机制 |
| TITLE： | Effects of Sulforaphane on the Proliferation and Apoptosis of Human Renal Tubular Epithelial Cells Induced by High Glucose and Its Mechanism |
| 摘要： | 目的：研究萝卜硫素对高糖诱导人肾小管上皮细胞HK-2增殖和凋亡的影响，并初步探讨其作用机制。方法：将HK-2细胞分为正常组、高糖组、厄贝沙坦组（阳性对照，1μmol/L）和萝卜硫素低、中、高浓度组（10、20、40μmol/L），正常组细胞用DMEM完全培养基培养96h，其余各组细胞均用高糖DMEM完全培养基（含40mmol/L葡萄糖）培养48h诱导细胞损伤后，再加入相应药物继续培养48h。检测各组细胞的存活率、凋亡率以及细胞中细胞周期蛋白D1（cyclinD1）、胱天蛋白酶3（caspase-3）、B细胞淋巴瘤2（Bcl-2）、B细胞淋巴瘤2相关X蛋白（Bax）mRNA和磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）、磷酸化腺苷一磷酸活化蛋白激酶（p-AMPK）、磷酸化蛋白激酶B（p-Akt）、磷酸化磷脂酰肌醇3激酶（p-PI3K）蛋白的表达量。另将HK-2细胞分为正常组、高糖组、萝卜硫素高浓度组（40μmol/L）、阿卡地新组（AMPK激动剂，1mmol/L）、萝卜硫素高浓度+compoundC组（萝卜硫素40μmol/L+AMPK抑制剂compoundC40μmol/L）、哌立福辛组（Akt抑制剂，19.95μmol/L）、萝卜硫素高浓度+SC79组（萝卜硫素40μmol/L+Akt激动剂SC794μmol/L），同法培养后，检测细胞中p-mTOR、p-AMPK、p-Akt、p-PI3K蛋白的表达量。结果：与正常组比较，高糖组细胞的存活率以及细胞中cyclinD1、Bcl-2mRNA和p-AMPK蛋白的表达量均显著降低（P＜0.05），凋亡率以及细胞中cas-pase-3、BaxmRNA和p-mTOR、p-Akt、p-PI3K蛋白的表达量均显著升高（P＜0.05）。与高糖组比较，萝卜硫素低、中、高浓度组和厄贝沙坦组细胞上述指标均显著改善（P＜0.05），其中萝卜硫素中、高浓度组细胞上述指标的改善效果均显著优于萝卜硫素低浓度组（P＜0.05）。与厄贝沙坦组比较，萝卜硫素高浓度组细胞上述指标差异均无统计学意义（P＞0.05）。与萝卜硫素高浓度组比较，阿卡地新组细胞中p-AMPK、p-mTOR蛋白和哌立福辛组细胞中p-mTOR、p-Akt、p-PI3K蛋白的表达量差异均无统计学意义（P＞0.05）；萝卜硫素高浓度+compoundC组细胞中p-AMPK蛋白的表达量显著降低（P＜0.05），p-mTOR蛋白的表达量显著升高（P＜0.05）；萝卜硫素高浓度+SC79组细胞中p-mTOR、p-Akt、p-PI3K蛋白的表达量均显著升高（P＜0.05）。结论：萝卜硫素可促进高糖诱导肾小管上皮细胞增殖，抑制其凋亡，其机制可能与上调p-AMPK表达，下调p-mTOR、p-Akt、p-PI3K表达有关。 |
| ABSTRACT： | 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. |
| 期刊： | 2021年第32卷第24期 |
| 作者： | 周磊，艾望，陈小娟，闻春月 |
| AUTHORS： | ZHOU Lei，AI Wang，CHEN Xiaojuan ，WEN Chunyue |
| 关键字： | 萝卜硫素；高糖；人肾小管上皮细胞；磷脂酰肌醇3激酶/蛋白激酶B信号通路；腺苷一磷酸活化蛋白激酶/哺乳动物雷帕 |
| KEYWORDS： | Sulforaphane；High glucose ；Human renal tubular epithelial cells ；PI3K/Akt signaling pathway ；AMPK/mTOR |
| 阅读数： | 223 次 |
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