失憶型輕度認知功能障礙（a-MCI）患者被認為是演變成為阿茲海默症的高危險群，在早期由於海馬迴結構的神經纖維糾結，患者會開始產生記憶障礙。Braak和Braak在1991年將神經纖維糾結分成六個時期，開始會先從海馬迴結構的前內鼻區和側海馬迴產生病變，最後才會順延到海馬迴本體。由於前內鼻區主要處理事件及物件特徵，側海馬迴主要處理空間背景訊息，海馬迴本體主要處理空間位置記憶，所以本研究假設事件及物件特徵與空間背景訊息的配對記憶在a-MCI階段就會產生障礙，而空間位置記憶則在輕度阿茲海默症會開始產生障礙，如果不同階段神經病變的認知功能表現，可以在研究結果中呈現出來，或許可以協助找到早期偵測海馬迴結構病變的神經認知功能指標。 本研究受試者主要包含正常組（NC組）30人，失憶型輕度認知功能障礙組（a-MCI組）30人和輕度阿茲海默症組（AD組）30人，共計90人。每組受試者均接受神經心理測驗衡鑑和本研究自行發展的空間脈絡記憶測驗。空間脈絡記憶測驗總共分為三個部分：（1）空間位置記憶測驗：要求受試者回憶之前在地圖上隨機出現的建築物位置；（2）事件與地點連結測驗：事件和地點配對出現後，要求受試者選擇事件所配對的地點背景為何；（3）地點與物體的連結測驗：物體和地點配對出現後，要求受試者選擇該地點之前出現的物體為何。 研究結果呈現，不同組別在神經心理測驗結果，a-MCI組在延宕提取以及記憶保留的部分相較於其他認知功能顯著較差，而AD組相較於a-MCI組，除了記憶力表現更差外，其他認知功能的缺損也更為嚴重。而不同組別在空間脈絡記憶的結果，空間位置記憶分測驗呈現NC組＞a-MCI組＞AD組的結果，在事件與地點的連結分測驗呈現NC組＞a-MCI組＝AD組的結果，在地點與物體的連結分測驗呈現NC組＞a-MCI組＞AD組的結果。從ROC曲線分析呈現，空間脈絡記憶測驗相較於其他篩檢測驗，在區分NC組和a-MCI組的敏感度及特異度較好，而MMSE則在區分a-MCI組和AD組的敏感度及特異度較好。 研究結果呈現a-MCI受試者在一般認知功能尚未顯著下降的同時，空間脈絡記憶就已經呈現障礙，這可能和早期神經纖維糾結所破壞的區域有關，結果也呈現空間脈絡記憶測驗在a-MCI階段，比其他篩檢測驗能更敏感的區分出正常和異常的患者。目前臨床常用的MMSE測驗，因為複合了多項認知功能，反而適合用於篩檢已經為輕度阿茲海默症的患者。 / Background: Amnesic mild cognitive impairment (a-MCI) was identified to have a high risk to become Alzheimer’s disease (AD). In early stage of AD, because of neurofibrillary tangles, patient began complaining progressive memory deficits. The progressive course of neurofibrillary tangles was divided into 6 stages (Braak and Braak, 1991). Initially, the neurofibrillary tangles destroyed perirhinal and parahippocampus neurons, which may correspond to the a-MCI stage and then proceed to hippocampal body that correspond to early AD. According to previous studies, the perirhinal is primarily associated with item features encoding, the parahippocampus associated with scene features encoding, and the hippocampus associated with spatial location memory. Based on these findings, we hypothesized that the item and scene features association memory would show impairments in a-MCI and the spatial location memory would not be impaired in a-MCI but in early AD. If the different stages could be discriminated by the performance on spatial context memory test that we design, it can be utilized in clinical settings to assist the diagnosis of a-MCI. Method: Three groups of subjects were selected from the clinic of the neurological department of Chang Gung Memorial Hospital, including normal subjects (n=30), a-MCI subjects not diagnosed with dementia (n=30), and mild AD subjects (n=30). All of them were administered a package of neuropsychological tests and a self-developed spatial context memory test that include three sub-tests: (1) a spatial location memory test: subjects have to recognize the location of a building that was appeared in a map; (2) an event-place association memory test: subjects need point out which spatial scene that was associated with this event; and (3) a place-object association memory test: subjects need point out which object that was associated with this place shown before. Result: In neuropsychological tests, a-MCI group demonstrated significant impairment in delay retrieval and memory retention in comparison to their performance on tests for other cognitive functions. The AD group showed decline in overall cognitive functions including declarative memory and others. In the spatial context memory test, both the spatial location memory test and the place-object association memory subtest showed a decline in a-MCI group, and a further decline in AD group; the event-place association memory test presented significant decreases in both a-MCI and AD group in comparison to normal control, but no difference between the two clinical groups. Conclusion: The current study shows that the spatial context memory in a-MCI patients has greater impairment than their general cognitive function. Compared with other screening test, the spatial context memory has greater sensitivity and specificity to discriminate a-MCI from NC.

中文摘要 厚朴，採用厚朴植物之樹皮，是ㄧ種已知可應用於治療精神疾病的傳統天然藥物，例如：憂鬱症等。厚朴主要的有效多酚環成分已被證實具有抗氧化、抗發炎及抗興奮性毒殺等神經保護作用，因此，推測厚朴可作為一種潛在治療像是帕金森氏症這累神經退化性疾病之藥物。本研究之目的為探討厚朴是否可以預防與治療因百草枯及MPTP所誘導的毒害及學習、記憶和運動功能缺失等行為異常現象。本研究監測Oregon-R品系之果蠅(年齡：1-2, 20天或30天)之壽命在長期暴露於百草枯(5-20 mM)並先給予厚朴(100, 300或600 mg/L)治療之變化。其結果顯示，厚朴無法延長暴露在百草枯環境下之果蠅壽命。另外，我們給予雄性ICR小鼠(30-35 g)，連續五天，每日一劑MPTP(25 mg/kg, i.p.)，誘導神經毒性及行為異常現象。在共同投藥組別，在給予MPTP注射前一小時，先以灌餵方式給予小鼠厚朴(100或300 mg/kg)預防，連續五天後，只單獨給予厚朴治療連續十四天。後投藥組別，在給予最後一劑MPTP後，連續十四天給予厚朴(100或300 mg/kg).治療。在控制組別中，給予生理食鹽水(0.9%, i.p.)及灌餵玉米油。結果顯示，MPTP與厚朴並不影響小鼠之運動協調功能，然而，可利用新位置辨識能力測試及新物體辨識認知行為測試，檢測因MPTP所引起之認知功能障礙現象，由我們結果中顯示，不論是與MPTP共同給予厚朴治療抑或是後處理厚朴皆可恢復因MPTP所造成的認知功能障礙現象，此外，厚朴也可恢復因MPTP所造成多巴胺神經元及多巴胺轉運子受損之情形，另外，我們也初步發現，厚朴可在海馬迴中使Nrf2表現量提升。因此，初步結果表明，厚朴將可成為未來治療帕金森氏症之天然藥物。 / Cortex Magnoliae, the bark of Magnolia officinalis, has been prescribed in the traditional herbal medicine to treat a variety of mental disorders including depression. The main constituents of cortex Magnoliae contain the biphenyl compounds such as honokiol and magnolol. Both biphenyl compounds were shown to have the neuronal protective effect which is related to the anti-oxidation, anti-inflammation, and anti-excitatory toxicity. Thus, it was proposed that cortex Magnoliae may act as the potential therapeutic agent for the treatment of neurodegenerative disorders such as Parkinson’s disease (PD). The aim of the present study was to examine whether cortex Magnoliae exhibits the neuroprotective and therapeutic action against the neuronal toxicity and behavioral deficits in learning, memory, and motor function induced by neurotoxin paraquat and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in PD-like models. The lifespan of flies from Oregon-R strain of Drosophila melanogaster (age: 1-2, 20 or 30 days) chronically exposed to paraquat (5-20 mM) with pre-treatment of Cortex Magnoliae (100, 300 or 600 mg/L) were measured. Results showed that pre-treatment of Cortex Magnoliae could not extend the lifespan of the flies reduced by paraquat. On the other hand, male ICR mice (30-35g) were administered with MPTP (25 mg/kg, i.p.) once daily for 5 consecutive days to induce neurotoxicity and behavioral impairment. In co-treatment group, male mice were orally administrated with cortex Magnoliae (100 or 300 mg/kg) 1 hour before MPTP injection for 5 days and then followed by oral administration of cortex Magnoliae alone for consecutive 14 days. Mice in post-treatment group were orally administered with cortex Magnoliae (100 or 300 mg/kg) for consecutive 14 days after the final injection of MPTP. Mice in control group were injected with saline (0.9%, i.p.) and orally administrated with vehicle (corn oil). Our results showed that MPTP and cortex Magnoliae did not affect mouse coordination and balance in beam walking test. However, cortex Magnoliae improved the cognitive impairments determined by novel-location recognition task (NLRT) and novel-object recognition task (NORT) in MPTP-induced PD mouse. Additionally, cortex Magnoliae restored MPTP-induced loss of dopaminergic neurons and recovered MPTP-induced loss of dopamine transporters in striatum. Cortex Magnoliae also activated Nrf2 in hippocampus. Therefore, the preliminary results suggest that cortex Magnoliae may be a novel candidate for the treatment of Parkinson's disease in the future. The pharmacological mechanism of cortex Magnoliae in PD treatment needs further study.

雙酚化合物在文獻報導中發現具有抗發炎和抗氧化的能力，因為其親脂性的特性，雙酚化合物可以輕易穿透血腦屏障到中樞神經系統發揮其藥理活性。因此，雙酚化合物被評估可做為潛在預防及治療神經退化性疾病如帕金森氏症的神經保護藥物。本研究目的為探討新合成的雙酚化合物MH101及MH102是否具有神經保護和治療效用，而對抗神經毒素(包含巴拉圭、過氧化氫及MPTP)引起的神經毒害及其誘發的動物行為異常(如: 學習、記憶及運動協調)。研究中應用Oregon-R的果蠅(年齡: 1-2, 7, 20 和 30天)做為檢測模式，果蠅暴露在巴拉圭 (5-20 mM)或過氧化氫(0.3 %-3 %)環境下，並且給予MH101 (0.1-3 μM)。結果顯示MH101未能有效地減緩巴拉圭及過氧化氫所引起果蠅壽命的下降。此外，給予雄性ICR小鼠 (25-30 g) 腹腔注射MPTP (25mg/kg)每天一次連續五天，觀察神經毒素誘發的行為異常和神經毒害。在觀察保護效果的研究中，雄性小鼠在給予MPTP前一小時腹腔注射MH101 (1-3 mg/kg) 或MH102 (0.1-3 mg/kg) 每天一次連續五天，之後單獨給予MH101或MH102治療連續九天。後處理的組別，雄性小鼠在給予MPTP每天一次連續五天後，每天腹腔注射MH101 (1-3 mg/kg) 或MH102 (0.1-3 mg/kg)連續九天。控制組組別，小鼠則給予生理食鹽水(0.9%)及玉米油的混合液。結果顯示，MH101、MH102及MPTP皆不影響小鼠橫桿行走試驗的運動平衡和協調能力。然而，在前處理和後處理MH101或MH102後用新位置辨識能力測試和新物體辨識能力測試觀察MPTP引起的認知缺失，實驗結果顯示MH101及MH102皆恢復短期記憶和長期記憶的認知辨識指標。另外，前處理和後處理MH101或MH102雖有些微恢復紋狀體內MPTP引起多巴胺神經損傷及多巴胺轉運子減少的趨勢，但不顯著。由此推論，雙酚化合物MH101及MH102具有預防及改善神經毒素所引發的認知與學習缺陷，未來可能發展成為神經退化性疾病如帕金森氏症之潛力治療藥物，另針對MH101及MH102在神經損傷及動物行為障礙的恢復和保護藥理機制則需進一步實驗探討。 / Biphenols which are the main constituents of the traditional herbs have been found to possess the antiinflammatory and antioxidative properties. Due to the lipophilic activity, biphenols can readily cross the blood brain barrier to exert their pharmacological effects in the central nervous system. Thus, biphenols are proposed to act as the novel neuroprotective agents for treatment of neurodegenerative disorders such as Parkinson’s disease (PD). The aim of the present study was to examine whether the new synthetic biphenolic compounds MH101 and MH102 have the neuroprotective and therapeutic actions against the neurotoxicity and the behavioral impairments (e.g. learning, memory, and motor coordination) induced by neurotoxins including paraquat, hydrogen peroxide, and MPTP in PD-like animal models. The following experiments examined the lifespan of flies from Oregon-R strain of Drosophoila melanogaster (age: 1-2, 7, 20 and 30 days) chronically exposed to paraquat (5-20 mM) or hydrogen peroxide (0.3 %-3 %) under MH101 (0.1-3 μM) treatment. Our results showed that MH101 could not effectively influence the reduced lifespan of the flies induced by paraquat and hydrogen peroxide. Furthermore, male ICR mice (25-30 g) were administrated with MPTP (25 mg/kg, i.p.) once daily for 5 consecutive days to induce neuronal damage and cognitive deficits. For the protective study, male mice were administrated with MH101 (1-3 mg/kg, i.p.) or MH102 (0.1-3 mg/kg, i.p.) 1 hour prior to MPTP injection once daily for 5 days, and followed daily treatment with MH101 or MH102 alone for consecutive 9 days after the final injection of MPTP. For the post-treatment study, male mice were administrated with MPTP (25 mg/kg, i.p.) once daily for 5 consecutive days, and followed by daily treatment of MH101 or MH102 for 9 days. Mice in control group were injected with vehicle (0.9% saline + corn oil). The results showed that MH101, MH102, and MPTP alone did not alter the motor functions of coordination and balance in beam walking test. On the other hand, both pre-treatment and post-treatment of MH101 and MH102 reversed the cognitive dysfunction induced by MPTP detected by novel location recognition test (NLRT) and novel object recognition test (NORT). Data demonstrated that MH101 and MH102 reversed the reduction in recognition index (RI) of short term memory and long term memory in MPTP-induced PD model. However, pre-treatment and post-treatment of MH101 or MH102 slightly recovered MPTP-induced loss of dopamine neurons and dopamine transporter in striatum. Therefore, the results suggest that biphenols including MH101 and MH102 may be the candidates for treatment of neurodegenerative diseases such as PD. In the future, it will need further study to determine the pharmacological mechanism of MH101 and MH102 in protection and restoration of neuronal injury and cognitive impairment.