[Effect of salvianolic acid B on high-glucose induced renal tubular epithelial-mesenchymal transition in rats and its mechanism].
[Effect of salvianolic acid B on high-glucose induced renal tubular epithelial-mesenchymal transition in rats and its mechanism].
复制标题
丹酚酸B对高糖诱导大鼠肾小管上皮间质转化的影响及其机制
DOI：
10.19540/j.cnki.cjcmm.20200405.403
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
["Lan Sun
中科院分区：
文献类型：
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作者：
["Lan Sun
观察丹酚酸B(SalB)对高糖诱导的大鼠肾小管上皮-间质转化(EMT)的影响，探讨其防治糖尿病肾病的可能机制。体外培养大鼠肾小管上皮细胞NRK-52E。分为对照组、高糖组、高糖+10μmol.L-1SalB组(SalB组)、高糖+不同浓度(1、5、10μmol.L-1)SalB组、高糖+5.0μmol.L-1吡格列酮组、高糖+10μmol.L-1SalB+5μmol.L-1GW9662组，分别于6、12、24、48h进行动态观察。免疫印迹法检测PPARγ、PTEN、α-SMA、E-钙粘蛋白和PI3K/Akt信号分子的蛋白表达水平。实时定量聚合酶链式反应检测PPAR、γ和PTENmRNA的表达。用四甲基偶氮唑盐比色法测定NRK52 E细胞的存活率。结果表明，与对照组相比，高糖组PPARγ和PTENmRNA及蛋白表达水平逐渐降低，α-SMA和p-Akt308蛋白表达水平逐渐升高，E-钙粘蛋白蛋白表达逐渐降低(P<；0.05)。与高糖组比较，随着丹参B剂量的增加，高糖+不同浓度丹参B组PPARγ、PTEN的表达水平逐渐升高，α-SMA和p-Akt~(Thr308)的蛋白表达逐渐降低，E-钙粘蛋白的蛋白表达逐渐升高(P<；0.05)，而1μ·L浓度对PPARγ和PTEN的影响无显著差异。与高糖组比较，高糖+10γ·L-1Sal B动态观察组PPAR PTEN m RNA(除6 h外)、PTEN m RNA(除6 h外)及蛋白(除6、12 h外)的表达水平持续升高，α-SMA和p-Akt(Thr308)的蛋白表达水平(除6 h外)继续降低，E-μ蛋白的表达水平持续升高(P<；0.05)。与高糖组比较，丹参B组和吡格列酮能显著增强PPARγ、PTEN的表达，增强E-钙粘蛋白的蛋白表达，降低α-SMA、p-Akt~(308)蛋白的表达，但两组间差异无统计学意义(P>0.05)。而Sal B+GW9662对照组PPARγ、PTEN mRNA和蛋白、E-钙粘蛋白、α-SMA和p-Akt(Thr308)蛋白的表达水平与高糖组比较差异无统计学意义。PPARγ拮抗剂GW9662可阻断SAL B的作用。结论：丹参B对高糖EMT诱导的NRK52 E细胞有抑制作用。其机制可能与SALB激活PPARPTEN，上调γ表达，从而抑制PI3K/AKT信号通路的纤维化效应有关。
The aim of this paper was to observe the effect of salvianolic acid B(Sal B) on high-glucose induced renal tubular epithelial-mesenchymal transition(EMT) in rats, and to explore its possible mechanisms of prevention and treatment of diabetic nephropathy. The rat renal tubular epithelial NRK-52 E cells were cultured in vitro. The cells were divided into control group, high glucose group, high glucose+10 μmol·L~(-1)Sal B group(Sal B), the above 3 groups were set at 6, 12, 24 and 48 h for dynamic observation; high glucose+Sal B different concentration(1, 5, 10 μmol·L~(-1)) groups, high glucose+5.0 μmol·L~(-1) pioglitazone group, high glucose+10 μmol·L~(-1)Sal B+5 μmol·L~(-1)GW9662 group. The protein expression levels of PPARγ, PTEN, α-SMA, E-cadherin and PI3 K/Akt signaling molecules were determined by Western blot. The mRNA expression of PPARγ and PTEN were detected by Real-time PCR. The viabi-lity of NRK52 E cells was determined by MTT assay. The results showed that as compared with control group, the mRNA and protein expression levels of PPARγ and PTEN in high glucose group gradually reduced, the protein expression levels of α-SMA and p-Akt~((Thr308))gradually increased, and the protein expression of E-cadherin gradually reduced(P<0.05). As compared with high glucose group, when increases in Sal B doses, the mRNA and protein expression levels of PPARγ, PTEN in high glucose + different concentrations of Sal B groups gradually increased, the protein expression levels of α-SMA and p-Akt~((Thr308)) gradually reduced, and the protein expression of E-cadherin gradually increased(P<0.05), however, the effect of 1 μmol·L~(-1)concentration of Sal B on the expression of PPARγ mRNA and protein and PTEN mRNA was not significantly different. As compared with high glucose group, the mRNA and protein expression levels of PPARγ mRNA(except 6 h) and protein(except 6 h), PTEN mRNA(except 6 h) and protein(except 6, 12 h) kept increasing, the protein expression levels of α-SMA and p-Akt~((Thr308))(except 6 h) continued to reduce, the protein expression of E-cadherin kept increasing in high glucose+10 μmol·L~(-1) Sal B dynamic observation group(P<0.05). As compared with high glucose group, Sal B and the pioglitazone(PIO) can greatly enhance the expression of PPARγ, PTEN at mRNA and protein levels, enhance the expression of E-cadherin at protein levels, and reduce the expression of α-SMA, p-Akt~((Thr308))protein level(P<0.05), there was no significant difference between the two groups. However, the expression levels of PPARγ and PTEN mRNA and protein, E-cadherin, α-SMA and p-Akt(Thr308) protein in the Sal B+GW9662 control group were not statistically significant compared with the high glucose group. The effect of Sal B was blocked by the PPARγ antagonist GW9662. It can be concluded that Sal B can suppress the NRK52 E cells induced by high-glucose EMT. The mechanism may be related to the activation of PPARγ with Sal B, and the up-regulation of PTEN expression, and thereby inhibiting the fibrosis effect of PI3 K/Akt signaling pathway.