Qu Ling, Zhang Hong, Gu Bei, Dai Wei, Wu Qun-li, Sun Lian-qing, Zhao Li, Shi Yue, Liang Xiao-chun
Department of Traditional Chinese Medicine, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, 100730, China.
Cell Center, Institute of Basic Medical Science, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, 100005, China.
Chin J Integr Med. 2016 Mar;22(3):185-92. doi: 10.1007/s11655-015-2164-8. Epub 2015 Mar 31.
To observe the deregulation of autophagy in diabetic peripheral neuropathy (DPN) and investigate whether Jinmaitong ( JMT) alleviates DPN by inducing autophagy.
DPN models were established by streptozotocin-induced diabetic rats and Schwann cells (SCs) cultured in high glucose medium. The pathological morphology was observed by the improved Bielschowsky's nerve fiber axonal staining and the Luxol fast blue-neutral red myelin staining. The ultrastructure was observed by the transmission electron microscopy. Beclin1 level was detected by immunohistochemistry and Western blot. The proliferation of cultured SCs was detected by methylthiazolyldiphenyl-tetrazolium bromide.
Diabetic peripheral nerve tissues demonstrated pathological morphology and reduced autophagic structure, accompanied with down-regulation of Beclin1. JMT apparently alleviated the pathological morphology change and increased the autophagy [in vivo, Beclin1 integral optical density (IOD) value of the control group 86.6±17.7, DM 43.9±8.8, JMT 73.3 ±17.8, P<0.01 or P<0.05, in vitro Beclin1 IOD value of the glucose group 0.47±0.25 vs the control group 0.88±0.29, P<0.05]. Consequently, inhibition of autophagy by 3-methyladenine resulted in a time- and concentration-dependent decrease of the proliferation of SCs (P<0.05, P<0.01).
Down-regulation of autophagy in SCs might contribute to the pathogenesis of DPN. JMT alleviates diabetic peripheral nerve injury at least in part by inducing autophagy.
观察糖尿病周围神经病变（DPN）中自噬的失调情况，并研究金脉通（JMT）是否通过诱导自噬来减轻DPN。
通过链脲佐菌素诱导的糖尿病大鼠和在高糖培养基中培养的雪旺细胞（SCs）建立DPN模型。采用改良的 Bielschowsky 神经纤维轴突染色和 Luxol 固蓝-中性红髓鞘染色观察病理形态。通过透射电子显微镜观察超微结构。采用免疫组织化学和蛋白质免疫印迹法检测 Beclin1 水平。用甲基噻唑基二苯基四氮唑溴盐检测培养的SCs的增殖情况。
糖尿病周围神经组织呈现病理形态且自噬结构减少，同时伴有 Beclin1 的下调。JMT 明显减轻了病理形态变化并增加了自噬[体内，对照组 Beclin1 积分光密度（IOD）值为 86.6±17.7，糖尿病组为 43.9±8.8，JMT 组为 73.3±17.8，P<0.01 或 P<0.05；体外，葡萄糖组 Beclin1 IOD 值为 0.47±0.25，对照组为 0.88±0.29，P<0.05]。因此，3-甲基腺嘌呤抑制自噬导致SCs增殖呈时间和浓度依赖性下降（P<0.05，P<0.01）。
SCs 中自噬的下调可能有助于 DPN 的发病机制。JMT 至少部分通过诱导自噬减轻糖尿病周围神经损伤。
