The protective effect of Cordycepin on diabetic nephropathy through autophagy induction in vivo and in vitro
The protective effect of Cordycepin on diabetic nephropathy through autophagy induction in vivo and in vitro
复制标题
虫草素诱导自噬对糖尿病肾病的保护作用
DOI：
10.1007/s11255-019-02241-y
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发表时间：
2019-10-01
影响因子：
2
通讯作者：
Wan, Qijun
中科院分区：
文献类型：
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作者：
Cao, Tao;Xu, Ricong;Wan, Qijun
糖尿病肾病（DN）是糖尿病（DM）最严重的慢性并发症之一。自噬是足细胞维持细胞内环境稳定的重要生理功能。在本研究中，我们计划阐明中药虫草素对DN的作用及其机制。方法将所有大鼠随机分为正常对照组、糖尿病对照组、低剂量组（10 mg/kg）、中剂量组（100 mg/kg）、高剂量组（500 mg/kg）。通过自动生化分析仪检测胆固醇、血糖、甘油三酯、肌酐和尿蛋白水平。采用酶联免疫吸附法（Elisa）检测IL-1 β、IL-6、IL-18水平。HE染色检查组织病理变化。TUNEL染色检测细胞凋亡。western blot检测纤维化标志物α - sma、t-TG、TIMP-1、凋亡相关蛋白cleaved-caspase3、Bax、Bcl-2、自噬标志物Beclin1、轻链3 (LC3)I/II、p62的表达。结果糖尿病对照组的胆固醇、血糖、甘油三酯、肌酐、尿蛋白水平均明显高于正常对照组。DN模型组也有明显的组织病理学损伤。虫草素治疗后，与DN组比较，上述指标均有改善，组织损伤也有所减轻。进一步研究表明，冬虫夏草素能抑制DN大鼠模型中细胞凋亡和肾纤维化，恢复细胞自噬。此外，我们的体外实验结果表明，添加3-甲基腺嘌呤（3-MA，特异性自噬抑制剂）通过诱导细胞凋亡和肾纤维化，成功地消除了冬虫夏草素对肾纤维化的保护作用。虫草素的上述保护作用呈剂量依赖性。结论冬虫夏草素参与了DN细胞凋亡、纤维化和自噬诱导的调节作用。我们的研究首次发现虫草素通过诱导自噬对大鼠DN有一定的治疗作用。
Background Diabetic nephropathy (DN) is one of the most serious chronic complications of diabetes mellitus (DM). Autophagy is an important physiological function for podocytes to maintain stability of intracellular environment. In this study, we planned to clarify the effect of Cordycepin, a traditional Chinese medicine, on DN and the related mechanisms. Methods All rats were randomly divided into normal control group, diabetic controls, low-dose group (10 mg/kg), medium-dose group (100 mg/kg), and high-dose group (500 mg/kg). The level of cholesterol, blood sugar, triglyceride, creatinine, and urine protein was examined through an automatic biochemistry analyser. Enzyme-linked immunosorbent assay (Elisa) was used to detect the level of IL-1 beta, IL-6, and IL-18. HE staining was used to examine histopathologic changes. TUNEL staining was used to detected cell apoptosis. The expression of fibrosis markers alpha-SMA, t-TG, and TIMP-1, apoptosis-related proteins cleaved-caspase3, Bax and Bcl-2, autophagy markers Beclin1, light chain 3 (LC3)I/II, and p62 were evaluated by western blot. Results The level of cholesterol, blood sugar, triglyceride, creatinine, and urine protein in the diabetic controls was much higher than that in the normal control group. Obvious histopathology injuries were also found in DN model group. After Cordycepin treatment, all the above indexes were improved compared with the DN group and tissue damages were also alleviated. Further studies showed that Cordycepin suppressed cell apoptosis and renal fibrosis and rescued cell autophagy in DN rat model. Moreover, the results of our in vitro experiments showed that the addition of 3-methyladenine (3-MA, specific autophagy inhibitor) successfully abolished the protective effect of Cordycepin on renal fibrosis through inducing apoptosis and renal fibrosis. The above protective effects of Cordycepin were exhibited in a dose-dependent manner. Conclusion Cordycepin participated in the modulation of cell apoptosis, fibrosis, and autophagy induction in DN. Our study for the first time revealed that Cordycepin had a certain therapeutic effect on DN in rats through autophagy induction.