中医调护PTSD大鼠"胎损"的行为及表观遗传机制研究

Post-traumatic stress disorder (PTSD) is a psychological traumatic incident, which, as the core symptom of PTSD, constantly presents and results in fear and anxiety. PTSD is extremely harmful to health. Our previous studies suggested the fear of maternal rats due to earthquake PTSD results in disorders of growth and behavior genetics, which could be released or corrected by rich environment and kidney-supplementing herbal medicine. Based on this, we planed to randomize 54 PTSD fear (PTSDf) rats into emotion group (EG), herbal medicine group (HMG), EG+HMG, and control group (CG). Environmental (physical and social simulation) and emotional enrichment will be applied in EG. In HMG kidney-yang supplementing will be given. EG+HMG refers to the combination of EG and HMG. Changes of behavior genetics e.g. growth and development will be observed and then high throughput chip technology will be employed to measure the expression and alteration of miRNA, and DNA methylation. Finally, targeted gene will be validated with rt-PCR. Thus, the differential expression of behavior related genes can be analyzed. We will integrate the development and growth of rat, behavior genetics, DNA methylation, neuro-hormones, miRNA to explore the genetics and epigenetics mechanism of rat fetus deficit resulting from PTSD fear, which is to say, tranditional Chinese medicine, biomedicine, bioinformatics, mathematics, molecular biology are synthetically applied.This project may provide scientific evidence and a new entry point to reveal the foundation of fetus defict and promote the development of TCM rehabilitation and TCM nursing.

创伤后压力心理障碍症（PTSD）的核心症状是恐惧焦虑，严重危害健康。前期自然科学基金项目研究表明，地震恐惧孕鼠会致子鼠生长发育与行为遗传障碍，优裕环境与补肾中药可有效改善或消除不良影响。故拟将PTSD恐惧孕鼠分为情志组、中药组、情志+中药组和空白组四组，分别给予环境调情志、中药温补肾阳和调情志与中药补肾同时干预；观测宏观行为与微观生物遗传指标，运用高通量芯片技术筛查候选miRNA，检测全基因组CpG岛甲基化水平，做差异基因转录因子活性检测和PCR验证。融合PTSD子鼠生长发育、行为遗传生物信息，集中西医、分子生物学等多学科分析，揭示表观遗传在PTSD发生、行为遗传、神经生物学变化等联系中发挥"桥接"的关键作用，阐释"恐伤肾"、"肾主生长发育"、"母病及子"等中医经典理论的科学内涵。为中医调护PTSD"胎损"提供科学依据和崭新视觉，促进中医养生康复和护理学发展。

本课题将72只初次受孕成功的SD雌性大鼠分成5组：PTSD组（n=12）、情志组（n=18）、中药组（n=12）、情志中药组（n=18）、正常对照组（n=12），且这5组孕鼠刚受孕时体重无统计学差异（P>0.05）。采用旷场试验、糖水试验、翻身测试、张耳睁眼测试对子鼠进行行为遗传检测。取第30日子鼠股动脉血进行表达谱芯片检测和甲基化测序，并进行关联分析。通过对SD孕鼠模拟PTSD恐惧“胎损”后采取优裕环境（即物理和社会环境）干预，以舒畅情志，模拟中医情志护理，同时给予温补肾阳的金匮肾气丸进行干预；观测宏观行为、神经激素、遗传效应与表达谱、表观遗传（甲基化）测试分析，筛选出中医调护对PTSD恐惧“胎损”可能发生表观遗传学修饰（甲基化异常）的基因，从而揭示中医调护PTSD大鼠恐惧“胎损”的行为遗传学与表观遗传学机制。. 母鼠孕期PTSD模拟可导致子鼠行为能力变差；中医情志调护与补肾中药可纠正母鼠PTSD导致的子鼠行为发育迟缓，情志联合中药的调护效果可能比单一干预的效果更好。PTSD可导致子鼠的5-HT、血清CORT、生长激素、多巴胺浓度异常，情志、中药干预可使这些异常恢复正常。母鼠孕期PTSD模拟可导致子鼠的多个基因表达谱异常，情志、中药、情志联合中药干预可从不同角度纠正这些异常。MAPK通路是情志和中药干预起效的共同基础，Ntf3与Pla2g5基因是MAPK信号通路的关键基因。母鼠孕期PTSD模拟导致的子鼠基因表达异常与其甲基化水平不同有关，表观遗传修饰可能是PTSD“胎损”及其情志、中药调护的关键机制。. 发表研究论文18篇，其中SCI收录论文2篇，核心期刊8篇。