Effect of osthole on advanced glycation end products-induced renal tubular hypertrophy and role of klotho in its mechanism of action
Effect of osthole on advanced glycation end products-induced renal tubular hypertrophy and role of klotho in its mechanism of action
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DOI：
10.1016/j.phymed.2018.09.030
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发表时间：
2019-02-01
期刊：
影响因子：
7.9
通讯作者：
Huang, Jau-Shyang
中科院分区：
文献类型：
--
作者：
Kan, Wei-Chih;Hwang, Jean-Yu;Huang, Jau-Shyang
背景资料：蛇床子素具有抗癌、抗炎、降血脂等药理活性。Klotho被鉴定为多种组织中的抗衰老蛋白。klotho的丧失与慢性肾脏疾病有关。目的：研究蛇床子素对晚期糖基化终产物（AGE）诱导的人肾小管上皮细胞klotho表达的影响，探讨蛇床子素和外源性klotho对抗AGE诱导的肾小管上皮细胞肥大的分子机制。方法：MTT法检测细胞活力。通过Western印迹法测量蛋白质表达。通过实时定量PCR分析mRNA水平。以肥大指数评价细胞肥大生长。结果：随着AGE浓度的增加，klotho合成呈剂量依赖性下降。蛇床子素显著增加AGE抑制的klotho mRNA和蛋白表达。蛇床子素和外源性klotho治疗显着减弱AGE诱导的Janus激酶2（JAK 2）-信号转导和转录激活因子1（STAT 1）和STAT 3激活。此外，蛇床子素和外源性klotho增加了细胞因子信号转导抑制因子1（SOCS 1）和SOCS 3的蛋白水平。蛇床子素和外源性klotho抑制p21（Waf 1/Cip 1）/collagen IV/cytoplasmic的表达、总蛋白含量和细胞大小，从而证实了蛇床子素和外源性klotho逆转AGE诱导的细胞肥大的能力。结论：我们发现蛇床子素通过诱导klotho的表达和抑制JAK 2-STAT 1/STAT 3信号通路来减轻AGE诱导的肾小管肥大。这些结果也表明，klotho可能是一个独特的分子靶点，用于治疗糖尿病肾病。
Background: Osthole has been widely reported to have pharmacological activities such as anti-cancer, anti-inflammation and anti-hyperlipidemic effects. Klotho was identified as an anti-senescence protein in a variety of tissues. Loss of klotho has been associated with chronic kidney disease. However, potential roles and molecular events for osthole and klotho in diabetic nephropathy remain unclear.Purpose: In the current study, we undertook to study the effect of osthole on klotho expression in advanced glycation end products (AGE)-cultured human renal proximal tubular cells, and to investigate the molecular mechanisms of osthole and exogenous klotho against AGE-induced renal tubular hypertrophy.Methods: Cell viability was elucidated by MTT assay. Protein expression was measured by Western blotting. mRNA level was analyzed by real-time PCR. Cellular hypertrophy growth was evaluated by hypertrophy index. Relative cell size was detected by flow cytometry.Results: We found that raising the ambient AGE concentration causes a dose-dependent decrease in klotho synthesis. Osthole significantly increased AGE-inhibited klotho mRNA and protein expression. Osthole and exogenous klotho treatments significantly attenuated AGE-induced Janus kinase 2 (JAK2)-signal transducers and activators of transcription 1 (STAT1) and STAT3 activation. Moreover, protein levels of suppressor of cytokine signaling 1 (SOCS1) and SOCS3 were augmented by osthole and exogenous klotho. The abilities of osthole and exogenous klotho to reverse AGE-induced cellular hypertrophy were verified by the observation that osthole and exogenous klotho inhibited p21(Waf1/Cip1)/collagen IV/RAGE expression, total protein content, and cell size.Conclusion: Consequently, we found that osthole attenuated AGE-induced renal tubular hypertrophy via induction of klotho expression and suppression of the JAK2-STAT1/STAT3 signaling. These results also showed that klotho might be used as a unique molecular target for the treatment of diabetic nephropathy.