蒙药珍宝丸促进亚急性脊髓损伤模型大鼠神经功能恢复的作用及机制研究

Spinal Cord Injury (SCI) is a clinical common disease, having high morbidity, long-term lack of effective treatment drug. This topic is based on many years of experience in treasure bolus clinical treatment and preliminary experimental study. It found that treasure pills have reduced damage and protected the spinal cord in the acute phase of SCI. Subacute rats with SCI model are used, by rt-pcr and Western blotting and immunohistochemical, cell chemistry, molecular biology methods such as SCI subacute period after application treasure bolus of serum MDA, SOD, GSH, TNF - a and organizations such as c - fos, the expression of BCL and other factors, more application treasures pills and the control group, the difference between validation treasures pills for the treatment of SCI in subacute stage. At the same time after the injury segment white matter of spinal cord injury in different periods are calculated separately, and the residual area, and the inclined plate test and BBB score observed in the rat hind limbs motor function recovery, contrast experiments to explore the treasure bolus promote the functional recovery of spinal cord function and mechanism, at the same time hope to discover regulatory factors associated with spinal cord injury and recovery, and analyzes the mutual relations. It provides a simple, cheap and effective treatment for SCI long-term treatment and rehabilitation.

脊髓损伤（Spinal Cord Injury，SCI）是临床常见疾患，致残率高，远期缺乏有效治疗的药物。本课题基于多年珍宝丸临床治疗经验及前期实验研究发现珍宝丸对SCI急性期有减轻损伤、保护脊髓功能的作用，采用亚急性大鼠SCI模型，通过RT-PCR和Western-blotting、免疫组织、细胞化学、分子生物学等方法检测SCI亚急性期应用珍宝丸后血清MDA、SOD、GSH、TNF-a等及组织中c-fos, bcl等因子的表达，比较应用珍宝丸和对照组之间的差异，验证珍宝丸对SCI亚急性期有治疗作用。同时在伤后分别计算不同时期损伤节段脊髓白质残存面积，以及采用斜板试验和BBB评分观察大鼠后肢运动功能的恢复情况，进行实验对照来探讨珍宝丸促进脊髓功能恢复的作用及机制，同时希望发现与脊髓损伤及恢复有关的调控因子，并分析其相互关系，能够为SCI的远期治疗及康复提供简便、价廉而行之有效的治疗方案。

背景：脊髓损伤（Spinal Cord Injury，SCI）是临床常见疾患，致残率高，远期缺乏有效治疗的药物。本课题基于多年珍宝丸临床治疗经验及前期实验研究发现，含珍珠，蟾蜍，麝香，羚羊的珍宝丸对脊髓损伤（SCI）有良好的治疗作用。 然而，其修复脊髓损伤的复杂机制尚不清楚，脊髓损伤中各种复杂的分子机制还不是十分明确，还需要进一步的探讨及研究。.主要研究内容：本实验从珍宝丸入手，使用脊髓损伤的大鼠模型进行研究，以期得到珍宝丸在脊髓损伤再生修复中的作用和相关机制。.重要结果：1.珍宝丸通过上调HSP27减少Treg细胞缓解大鼠脊髓损伤2.珍宝丸可通过上调miR-146a-5p表达来抑制GPR17的表达，进而抑制神经元细胞凋亡，促进ASCI大鼠脊髓功能的恢复3.珍宝丸通过lncRNA TUG1/miR-214/HSP27减少Treg细胞比例。.关键数据：1、珍宝丸通过HSP27抑制Treg细胞分化，通过抑制miR-214促进HSP27的表达。当珍宝丸的浓度2.5 mg/ml效果最明显。2. 经珍宝丸给药后，只有miR-146a-5p的表达发生了显著地变化，提示珍宝丸可能通过调节miR-146a-5p参与ASCI，过表达miR-146a-5p抑制野生型GPR17 3’UTR荧光素酶活性，且过表达miR-146a-5p下调SH-SY5Y细胞中GPR17的表达；而干扰miR-146a-5p呈现相反趋势。缺氧能下调SH-SY5Y细胞中miR-146a-5p的表达，上调GPR17的表达；而珍宝丸能逆转缺氧刺激的作用。珍宝丸可通过上调miR-146a-5p表达来抑制GPR17的表达，进而抑制缺氧诱导的SH-SY5Y细胞的凋亡。珍宝丸通过miR-146a-5p/GPR17促进ASCI大鼠脊髓功能的恢复。3. 珍宝丸下调脊髓损伤（SCI）大鼠中lncRNA TUG1的表达，LncRNA对miR-214的靶向调控作用，珍宝丸通过TUG1/miR-214调节HSP27表达和影响Treg细胞分化。.科学意义：部分阐明了珍宝丸在ASCI中的调控作用机制，为珍宝丸在ASCI临床应用提供了新的实验依据，同时为ASCI的防治提供了新的潜在作用靶点。