| 摘要: | 腦血管疾病，特別是中風，與精神疾病的高發生率有關。憂鬱症是中風後持續長時高風 險復發的疾病。儘管中風合併症的高發生率，中風後憂鬱症大多仍無法識別和治療。但重要 的是中風後憂鬱（Poststroke depression; PSD)，不但使腦中風患者的死亡率提高且阻礙身體康 復進入，更導致運動和認知功能退化，嚴重降低生活品質。其實腦中風和憂鬱症有許多共同 的病理標的，尤其是造成海馬迴神經再生(neurogenesis)的減少，因此產生海馬迴功能不足。 許多證據顯示：神經可塑性和内生性神經再生兩者扮演PSD病理上關鍵角色。本研究計畫目 的：我們想利用自發跑步運動合併使用補陽還五湯(BHD)來促進中風鼠神經再生以達到神經 保護和抗PSD作用，同時研究L1- INP細胞增生和腦部神經再生之可能信號傳導途徑和參與神 經增殖和再生的核心分子標靶/訊息及改變miRNAs表現情形而影響關鍵基因調控。我們的核 心假設是：自發跑步運動合併使用補陽還五湯可以減少“興奮神經毒性”，提升“神經再生” 來改善腦缺血損傷小鼠的中風後憂鬱(PSD)行為。這一假說是基於我們的先前已研究顯示：經 常跑步的小鼠可保護其減少受中風引起的腦損傷，延長其壽命；另外日口服補陽還五湯 (0.5-1.0克/kg體重)可減少中風產生的巨量自由基及發炎造成的血腦屏障破損滲漏，並促進內 生性神經再生及分化，得以改善腦梗塞和延長中風鼠壽命。本計畫研究理由是，自發跑步運 動合併使用補陽還五湯被認為具有神經保護作用已有很長一段時間，但在小鼠缺血中風導致 的興奮性神經毒性模型，探討自發跑步運動合併使用補陽還五湯兩個簡單的方法，是否可發 展成有效的中風後憂鬱症(PSD)治療策略(包括：生物標誌物和藥物靶標），以應用於缺氧/缺血 相關的疾病的治療。因此，我們的具體目標# 1 (第1年），為研究自發跑步運動合併使 用補陽還五湯是否可以藉由調節缺血中風小鼠Notch1信號增強海馬迴神經再生和L1-INP細 胞參與改善中風後憂鬱(PSD)。具體目標# 2 (第2年，研究自發跑步運動合併使用補陽還五 湯是否可以藉由活化缺血中風小鼠Wnt/p-catenin和Ang1/Tie2信號增強內生/海馬迴神經發 生。具體目標# 3 (第3年，自發跑步運動合併使用補陽還五湯是否可以藉由改變缺血中風 小鼠miRNAs表現而影響關鍵基因，增強血管生成促進血腦屏障再塑及神經再生。我們的預 期的結果：（1)可以揭示自發跑步運動合併使用補陽還五湯對缺血性中風引起的興奮性神經毒 性神經保護作用及抗PSD作用，是否可透過調節Notch1信號及密切關聯信息以促進NPC到 L1-INP細胞的生成或間接地干預有害信號（例如：p53，AP- 1)和增強保護信號（例如， CREB ，HSF- 1，BDNF，PI3K/Akt)等與GSK-3密切相關的訊號。（2)可以確定自發跑步 運動合併使用補陽還五湯治療是否能提高中風鼠內生/海馬迴神經再生及遷移並活化 Wnt/p-catenin和Ang1/Tie2信息參與神經生長因子(的Ang1和SDF1)表現，來解釋如何透過自發 跑步運動合併使用補陽還五湯治療可保護小鼠對抗中風後憂鬱症(PSD)，延長小鼠的壽命。這 些結果將十分有助於支持發展自發跑步運動合併使用補陽還五湯臨床上治療缺血型腦中風及 中風後憂鬱症(PSD)的有益策略。（3)可以提供證據來揭示自發跑步運動合併使用補陽還五湯 治療是否可以藉由改變miRNAs表現而影響關鍵基因以增BBB重塑，降低中風後缺血性血腦 屏障滲漏（BBB leakage)，並增加神經幹細胞增生、遷移、分化。總結，這些成果將可解釋自 發跑步運動合併使用補陽還五湯治療是否能藉由提高Notchl ’ Wnt/p-catenin，Ang1/Tie2等 信號和改變miRNAs表達以影響關鍵基因，藉以促進神經幹細胞增生、分化、遷移以修護中 風後神經傷害及抗PSD。綜合計畫結果，將可提供該策略(自發跑步運動合併使用補陽還五湯) 以及有效分子標靶及訊息機制，做為臨床發展成為治療中風及PSD的新穎療法。
Cerebrovascular diseases, especially stroke, are associated with a high incidence of psychiatric disorders. Depression is often persistent after stroke, with high risk of relapse even after remission over a long period of time. Despite the high level of comorbidity, depressive symptoms appear to remain frequently unrecognized and untreated. Most importantly, stroke patients with post-stroke depression (PSD) suffer higher mortality rates and show only minor improvement in rehabilitation programs, resulting in worse functional (motor and cognitive) outcomes and poorer quality of life. Decreased neurogenesis in the hippocampus via aging or stress has been implicated in the pathogenesis of cognitive deficits, anxiety and depression. Excitotoxicity is the major pathophysiological mechanism associated with stroke and PSD. Excitotoxicity induces inflammation and oxidative stress-associated disruption of the blood-brain barrier (BBB) to cause a great increase in neuronal cell death during ischemic stroke and the regeneration efficiency of neurons in the injured brain. General goal: to explore the neuroprotective effect and anti-psychiatric like disorder (e.g., post-stroke depression (PSD)) that has been associated with adult hippocampal neurogenesis by using voluntary running in combination with BHD treatment. Especially, we will focus on ⑴ endogenous/hippocampus neurogenesis and L1-INP cell proliferation through Notch1 signaling, (2) promotion of BBB remodeling by enhancing angiogenesis and growth factors (Ang1/Sdf1) through activating Wnt/p-catenin and Ang1/Tie2 signaling, (3) as well as enhancement of angiogenesis and neurogenesis through modulation of miRNA profiling by performing voluntary running in combination with BHD treatment in a depressive-like behavior (e.g., PSD) in ischemic stroke murine model. Our central hypothesis is that voluntary running and BHD can ameliorate “excitotoxicity” and enhance “hippocampus neurogenesis” to ameliorate post-stroke depression (PSD) in post cerebral ischemia injured mice. This hypothesis is based on our preliminary results showing that regular running protects mice against stroke-induced brain injury and extend the life span of mice with an ischemic stroke; and oral administration of BHD (0.5-1.0 g/kg) daily can reduce extraordinary amounts of free radicals, inflammation, BBB leakage and promote endogenous neurogenesis/differentiation to ameliorate brain infarction and extend their lifespan in mice with a stroke within 7 days. The rationale for this proposed research is that performing voluntary running and BHD treatment are two simple methods known to be neuroprotective for a long time and to interpret how performing voluntary running in combination with BHD treatment work on the ischemic stroke-mediated excitotoxicity in a murine model will lead to an effective therapeutic application (stroke biomarkers and drug targets) for hypoxia/ischemia-related disorders like poststroke depression (PSD). To achieve the research objective, three specific aims will be pursued: Specific Aim #1 (1st year), to study whether performing voluntary running and BHD treatment can enhance endogenous/hippocampus neurogenesis and L1-INP cell proliferation through Notch1 signaling in ischemic murine model. Expected Results of Specific Aim #1: We will identify whether performing voluntary running and BHD treatment can enhance endogenous/hippocampus neurogenesis, migration and differentiation of NPCs to L1-INP or Tbrl (glutamatergic cortical neuron) or TH (dopaminergic neurons) and the involvement of Notchl signaling in ischemic murine model to explain how performing voluntary running and BHD treatment could protect mice against PSD and extend the life span of mice with an ischemic stroke. These results could help support voluntary running and BHD treatment to be beneficial agents for the treatment of PSD in ischemic stroke. Specific Aim #2 (2nd year), to study whether performing voluntary running and BHD can promote BBB remodeling by enhancing angiogenesis and neural growth factors (Angl and Sdfl) through activating Wnt/p-catenin and Ang1/Tie2 signaling in ischemic murine model. Expected Results of Specific Aim #2: These data will provide evidence to reveal whether performing voluntary running and BHD treatment can decreases BBB leakage through increasing vascular integrity (BBB remodeling) via up-regulating Ang1/Tie2, Sdfl and tight junction protein (occludin) expression through activating Wnt/p-catenin in the ischemic brain after stroke, and will explain whether performing voluntary running and BHD treatment can increase neuroblast migration, which may be mediated by vascular remodeling through increasing expression of Ang1/Tie2, Sdfl or Wnt ligands, finally, can promote SVZ neuroblast migration. Specific Aim #3 (3rd year), to study whether performing voluntary running and BHD treatment can protect acute ischemic stroke mice by enhancing angiogenesis and neurogenesis through modulation of miRNA profiling within ischemic brain tissue and circulating serum of an acute ischemic murine model. Since miRNAs have the potential to serve as disease biomarkers as well as disease regulators. Therefore, it is important to explore the miRNAs in serum as well as in ischemic damaged tissue (brain) as biomarkers for PSD that will disclose the pathogenesis of PSD as well as prognosis biomarkers by performing voluntary running and BHD treatment. Expected Results of Specific Aim #3: These data will provide evidence to identify the changed expression pattern of miRNAs and the correlation with the common signaling pathway, e.g., Wnt/GSK-3/p-catenin in PSD mice (damage tissue and serum) as well as after by performing voluntary running and BHD treatment. Identification of serum and brain tissue differentially expressed miRNAs may also serve as potential diagnostic biomarkers for PSD and to demonstrate a novel role for miRNAs in the pathogenesis of PSD as well as prognosis markers by performing voluntary running and BHD treatment. |
