Department of Anesthesiology, The First Affiliated Hospital of Soochow University, Suzhou, China.
Department of Anesthesiology, Xuzhou Children's Hospital, Xuzhou Medical University, China.
Adv Clin Exp Med. 2021 Jun;30(6):623-632. doi: 10.17219/acem/131217.
Diabetic peripheral neuropathy (DPN) is one of the most common complications of diabetes, but the molecular mechanisms of DPN are still unclear.
To investigate the role of miR-221 in DPN and the related molecular mechanisms.
Streptozotocin (STZ) was used to establish an in vivo DPN model. An in vitro DPN model was established using high glucose-induced SH-SY5Y cells. The pain condition of rats was measured by evaluating the 50% paw withdrawal threshold (PWT) and paw withdrawal latency (PWL). Serum exosomes were extracted and identified. Expression of miR-221 in serum exosomes and serum SOCS3 expression were determined using reverse-transcription quantitative polymerase chain reaction (RT-qPCR). Western blotting was used to measure the protein levels of SOCS3, bradykinin (BK) and prostaglandin E2 (PEG2). The dual luciferase reporter assay was performed to confirm SOCS3 3'-UTR as a target of miR-221. The serum or cell supernatant levels of PEG2, BK, interleukin (IL)-6, IL-1β, and tumor necrosis factor alpha (TNF-α) were measured using enzyme-linked immunosorbent assay (ELISA).
Induction of the lenti-miR-221 inhibitor significantly decreased the expression of miR-221 in DPN rats. Both 50% PWT and PWL values were markedly decreased in DPN rats. When miR-221 was inhibited, the 50% PWT and PWL values were both significantly increased. Knockdown of miR-221 significantly increased the expression of SOCS3 and decreased the expression of NF-κB. Furthermore, knockdown of miR-221 remarkably decreased the expression of PEG2, BK, IL-6, IL-1β, and TNF-α in both STZ-treated DPN rats and high glucose-induced SH-SY5Y cells, which was reversed by inhibition of SOCS3. The dual luciferase reporter assay showed that miR-221 directly targeted and negatively regulated SOCS3.
Inhibition of miR-221 can reduce pain and decrease expression of inflammatory factors through targeting SOCS3 in DPN.
糖尿病周围神经病变（DPN）是糖尿病最常见的并发症之一，但 DPN 的分子机制仍不清楚。
探讨 miR-221 在 DPN 中的作用及其相关分子机制。
采用链脲佐菌素（STZ）建立体内 DPN 模型，高糖诱导 SH-SY5Y 细胞建立体外 DPN 模型。通过评估 50%足底撤回阈值（PWT）和足底撤回潜伏期（PWL）来测量大鼠的疼痛状况。提取血清外泌体并进行鉴定。采用逆转录定量聚合酶链反应（RT-qPCR）测定血清外泌体中 miR-221 的表达和血清 SOCS3 的表达。采用 Western blot 测定 SOCS3、缓激肽（BK）和前列腺素 E2（PEG2）的蛋白水平。采用双荧光素酶报告基因实验证实 SOCS3 3'-UTR 是 miR-221 的靶标。采用酶联免疫吸附试验（ELISA）测定血清或细胞上清液中 PEG2、BK、白细胞介素（IL）-6、IL-1β和肿瘤坏死因子-α（TNF-α）的水平。
诱导 lenti-miR-221 抑制剂可显著降低 DPN 大鼠 miR-221 的表达。DPN 大鼠的 50%PWT 和 PWL 值均明显降低。抑制 miR-221 后，50%PWT 和 PWL 值均明显升高。敲低 miR-221 可显著增加 SOCS3 的表达并降低 NF-κB 的表达。此外，在 STZ 处理的 DPN 大鼠和高糖诱导的 SH-SY5Y 细胞中，敲低 miR-221 可显著降低 PEG2、BK、IL-6、IL-1β和 TNF-α的表达，而 SOCS3 抑制可逆转这种作用。双荧光素酶报告基因实验表明，miR-221 可直接靶向并负调控 SOCS3。
抑制 miR-221 可通过靶向 SOCS3 减轻 DPN 大鼠的疼痛并降低炎症因子的表达。
