天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (4): 395-400.doi: 10.11958/20181506

• 实验研究 • 上一篇    下一篇

川芎嗪通过抑制PI3K/Akt/mTOR通路诱导自噬改善糖尿病肾病大鼠肾损害

钟娟 1，陈静 2，青姚 1，吴曙粤 1，钟庆荣 1△   

  1. 1南宁市第一人民医院（邮编 530022）；2南方医科大学南方医院
  • 收稿日期:2018-10-09 修回日期:2019-01-23 出版日期:2019-04-15 发布日期:2019-05-27
  • 通讯作者: 钟娟 E-mail:zjanny919@163.com
  • 作者简介:钟娟（1981），女，博士，副主任医师，主要从事中西医结合临床研究
  • 基金资助:
    川芎嗪对糖尿病肾病多元醇通路及P13k-Akt信号通路的实验研究;厄贝沙坦治疗高血压合并2型糖尿病患者脂肪肝的分子机制及临床研究

Experimental study of ligustrazine on improving renal damage in diabetic nephropathy rats by inhibiting the PI3K/Akt/mTOR pathway and inducing autophagy

ZHONG Juan1, CHEN Jing2, QING Yao1, WU Shu-yue1, ZHONG Qing-rong1△   

  1. 1 The First People’s Hospital of Nanning, Guangxi 530022, China; 2 Nanfang Hospital, Southern Medical University
  • Received:2018-10-09 Revised:2019-01-23 Published:2019-04-15 Online:2019-05-27
  • Contact: Juan Zhong E-mail:zjanny919@163.com

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摘要： 目的 探讨川芎嗪对糖尿病肾病（DN）大鼠肾脏 PI3K/Akt/mTOR信号通路和自噬标志蛋白 LC3B表达以及 尿微量白蛋白与尿肌酐比值（UACR）、肾脏病理的影响。方法 采用链脲佐菌素建立 DN大鼠模型，将模型大鼠随机 分为模型组，川芎嗪低、中、高剂量组，厄贝沙坦组；另设正常组，每组 12只。分别干预 8周后，采用酶法测定尿肌酐， 免疫比浊法测定尿微量白蛋白，计算 UACR；取肾组织，经甲醛固定后，进行苏木精-伊红（HE）和过碘酸-雪夫（PAS） 染色；通过蛋白免疫印迹法（Western blot）和免疫组化检测大鼠肾组织 PI3K/Akt/mTOR信号通路以及自噬标志蛋白 LC3B 表达的变化。结果 川芎嗪能减缓 DN 大鼠 UACR 的升高，改善其肾脏病理变化，其中川芎嗪中、高剂量组 UACR显著低于模型组（P＜0.05），且川芎嗪中、高剂量组与厄贝沙坦组间比较差异无统计学意义（P＞0.05）。此外， 川芎嗪能抑制 DN大鼠肾组织 p-PI3K、p-Akt、p-mTOR的表达，进而提高自噬标志蛋白 LC3B的表达水平和 LC3B-Ⅱ/ LC3B-Ⅰ比值。结论 川芎嗪能降低 DN大鼠 UACR的升高、改善其肾脏病理变化，发挥以上肾保护作用的机制可能 与其抑制 PI3K/Akt/mTOR信号通路，进而促进肾脏自噬有关。

关键词: 川芎嗪, 糖尿病肾病, 自噬, PI3K/Akt/mTOR通路, 厄贝沙坦, LC3B

Abstract: Objective To investigate the effects of ligustrazine (TMP) on the ratio of urine microalbumin to urinary creatinine (UACR), renal pathological changes, PI3K / Akt / mTOR signaling pathway and autophagosome marker protein LC3B in diabetic nephropathy (DN) rats. Methods The DN rat model was established by streptozotoein (STZ) peritoneal injection. DN model rats were randomly divided into model group, the low, middle and high dose of TMP groups, and irbesartan group. In addition, the normal rats were served as the normal group (n=12 for each group). After 8 weeks of intervention respectively, urine creatinine was determined by enzymatic method, urinary microalbumin was determined by immunoturbidimetry, and UACR was calculated. Kidney tissues were fixed with formaldehyde, hematoxylin-eosin (HE) and periodic acid-schiff (PAS) staining were performed. The protein expressions of p-PI3K, PI3K, p-Akt, Akt, p-mTOR, mTOR, and LC3B in the kidney tissues were detected by Western blot assay and immunohistochemistry (n=6 for each group). Results Compared with the model group, the increase of UACR was inhibited and the pathological changes in kidney were significantly ameliorated after treatment with TMP, especially in the middle and high dose groups, UACR were significantly lower than that of model group (P＜0.05). There was no significant difference in UACR between the middle and high dose groups and irbesartan group (P＞0.05). Moreover, the protein levels of p-PI3K, p-Akt and p-mTOR were decreased, while the expression of LC3B and the ratio of LC3B - Ⅱ/LC3B - Ⅰ were increased in TMP groups than those of model group. Conclusion TMP can reduce the increase of UACR and improve the pathological changes of kidney in DN rats. The molecular mechanisms that drive the therapeutic effects of TMP may involve in the inhibition of PI3K/Akt/mTOR signaling pathway to consequently promote renal autophagy.

Key words: Tetramethylpyrazine, diabetic nephropathies, autophagy, PI3K/Akt/mTOR pathway, irbesartan, LC3B