蒲金口服液对宫内感染/炎症致早产脑损伤大鼠神经髓鞘再生与修复的影响

Intrauterine infection/inflammation is the substantial cause of PVL. Children with PVL almost get the neuropsychiatric sequela, and the higher incidence of cerebral palsy, which is the main physical disability diseases in childhood. We previously present that 'phlegm' and 'stasis' is the main pathological element of brain damage in neonates . And we created Pu Jin Oral Liquid to treat children with hypoxic-ischemic encephalopathy (HIE) and the treatment results on HIBD is effective. Previous studies showed that Pu Jin Oral Liquid had the clear function of free radical scavenger, and could obviously decrease the level of NOS in the cerebral tissue of rats with hypoxic-ischemic encephalopathy, and the antagonism of platelet activating factor, which also have effectiveness on the treatment of premature PVL.It was supposed that 'phlegm' and 'stasis' is also the main pathological element of premature PVL caused by intrauterine infection/inflammation, and Pu Jin Oral Liquid can promote nerve myelin regeneration and repairation. This study will observe effects of different times which Pu Jin Oral Liquid bring about treating newborn rats with intrauterine infection or inflammation, and approach the expression of inflammatory cytokine and medullary sheath inhibitory factors, and its protective effect of oligodendrocyte and oligodendrocyte precursor cell from molecular biology aspect, thus it will lay theory foundations for prevention and treatment of activating microcirculation and removing stasis to PVL. This study may act an important role of preventing the happening and development of PVL, and lowering the mortality rate of cerebral palsy.

宫内感染/炎症是造成早产儿脑室周围白质软化（PVL）的重要原因。存在PVL的患儿几乎都有神经精神系统后遗损害，脑瘫发生率高，而脑瘫是儿童期主要的肢体残疾疾病。我们前期提出"痰"和"瘀"是新生儿脑损伤的主要病理因素，据此创制了"蒲金口服液"治疗小儿缺氧缺血性脑病（HIE），疗效显著，研究证实其具有明确的抗自由基作用，能显著降低缺氧缺血性脑损伤大鼠NOS含量、拮抗血小板活化因子，用于早产儿PVL的治疗亦有显效，推测痰瘀也是宫内感染/炎症所致早产儿PVL的主要病理因素，可促进神经髓鞘再生与修复。本研究拟观察不同时点使用蒲金口服液对宫内感染/炎症造成的早产鼠PVL的作用，并从分子生物学层面探讨其对影响髓鞘再生与修复的相关炎性细胞因子、髓鞘相关抑制因子、少突胶质细胞及其前体等的调控机制，为活血化瘀、化痰开窍药物早期干预治疗PVL奠定理论基础，对于防治PVL的发生、发展，降低脑瘫的发生率有重要意义。

脑性瘫痪(cerebral palsy，CP)是儿童神经残疾中最为严重和常见的疾病，宫内感染致早产引起的脑损伤是引起CP的常见高危因素。儿童脑室周围白质软化症（Periventricular Leuko-malacia，PVL）是造成早产儿脑瘫的主要原因。本病的治疗并无特效药物，需要多专业联合开展康复，且需终身康复，社会及家庭负担重。中药对于本病的早期干预治疗具有潜在的价值，“蒲金口服液”治疗小儿缺氧缺血性脑病（HIE）临床疗效显著。本项目是多年来对活血化瘀、化痰开窍中药临床研究与实践的基础上，系统研究蒲金口服液对宫内感染/炎症造成PVL大鼠神经再生与修复的影响。实验采用脂多糖（LPS）腹腔注射法制作宫内感染致早产鼠脑白质损伤模型，用不同剂量蒲金口服液及银杏叶提取物干预，评定脑损伤仔鼠神经行为学改变；电镜下观察其神经髓鞘厚度；并采用免疫组化、PCR、Western Blot等技术测定大鼠脑组织影响髓鞘再生与修复的相关因子及细胞凋亡情况。结果显示：.（1）宫内感染/炎症能引起早产脑损伤大鼠神经行为学改变，使脑组织中炎性细胞因子IL-6和TNF-α及髓鞘相关抑制因子Nogo-A、OMgp及NgR、p75NTR受体表达量上调，促进脑组织OL和OPC的凋亡。.（2）蒲金口服液干预可提高早产脑损伤仔鼠神经行为学能力；.（3）蒲金口服液对脑损伤仔鼠神经髓鞘厚度的影响，目前本部分已经完成预实验部分。正式实验完成造模取材，标本尚未完成制样、数据分析将后续完成。.（4）蒲金口服液可下调早产脑损伤仔鼠脑组织中IL-6和TNF-α的水平，减少髓鞘相关抑制因子Nogo-A、OMgp及NgR、p75NTR受体表达，抑制脑组织OL和OPC的凋亡。且早期干预相较晚期干预组差异有统计学意义，高剂量蒲金口服液组与低剂量组差异有统计学意义。.综上所述，蒲金口服液具有活血化瘀、化痰开窍之效，研究结果证明其可促进宫内感染/炎症致早产脑损伤大鼠神经再生与修复，支持了宫内感染/炎症致早产儿脑损伤的中医病理因素主要是“痰”和“瘀”，“痰瘀阻窍，经络不通”是其主要病机特点，活血化瘀、化痰开窍药物可促进宫内感染/炎症致PVL大鼠神经再生与修复，为蒲金口服液应用于临床提供了实验依据。