| Abstract: | Part I 研究目的: 頭部外傷將引發受損細胞或死亡細胞釋放各種調節因子並吸引微小膠細胞活化。活化狀態下的微小膠細胞將會改變其形態，並移行至受損區域釋放甲型腫瘤壞死因子及其他反應物質。本篇文章將探討，於液注撞擊所引發的頭部外傷過程中，給予傳統複方中藥MLC601是否可以藉由改善微小膠細胞的活化作用，達到降低損傷並促進運動功能的恢復。本篇文章使用MLC的原因在於 它對於腦中風治療有有顯著療效，並已經用以臨床實驗評估。 研究方法及資料：將動物隨機分成三組: 假手術組、頭部外傷給予安慰劑組及頭部外傷給予MLC601治療組。MLC給藥方式採用頭部外傷後一小時(早期)給予腹腔注射一劑MLC601(每公斤體重四毫克)，之後每一天注射一劑，連續三天。頭部外傷前及術後第四天(晚期)給予，連續評估其神經及運動功能缺陷。頭外部傷後第四天，使用免疫螢光染色及顯微鏡觀測神經元凋亡及微小膠細胞表達甲型腫瘤壞死因子於損傷區域表達之情形。免疫組織化學染色法用以觀察受損區域微小膠細胞之細胞數及型態。 研究結果：實驗顯示，於頭部外傷後會導致腦損傷、神經元凋亡、神經及運動功能缺陷、微小膠細胞聚集、微小膠細胞型態改變及過度表達甲型腫瘤壞死因子。以上這些現象，會因為早期MLC601的治療而得到較為明顯的改善。 結論與建議：傳統複方中藥MLC601可藉由調節微小膠細胞之變形及表達甲型腫瘤壞死因子達到大鼠頭部外傷之治療功效。 Part II 研究目的：探討於高山症發生前，先給予高壓氧前處理誘導熱休克蛋白質70產生，藉以保護及對抗高山症誘發之急性肺損傷及肺水腫機制。 研究方法及資料：將動物隨機分成四組，第一組:非高壓氧前處理及非高海拔壓力之控制組；第二組:非高壓氧前處理及誘發高海拔壓力組；第三組:高壓氧前處理及誘發高海拔壓力組；第四組:高壓氧前處理並給予熱休克蛋白質70抗體及誘發高海拔壓力組。高壓氧前處理是採取將動物至於2.0絕對大氣壓並供應100%氧氣濃度之高壓氧艙中一小時，連續五天。高海拔壓力誘導方式是將動物放置於0.47大氣壓(約6000公尺海拔高度，9.7%氧氣濃度)之低壓艙連續三小時。源自兔子的抗老鼠之熱休克蛋白質70多株抗體則於高海拔暴露前一小時以靜脈注射方式提供。當動物經過3小時高海拔壓力刺激後回復至常壓狀態時，即以過量麻醉劑進行安樂死並將動物器官移除進行組織及分子表現分析。 。 研究結果：非高壓氧前處理並誘發高海拔壓力組與控制組比較，有明顯的肺泡水腫、嗜中性白血球浸潤及肺泡出血之表現。此外，肺組織中第一型及第五型水通道蛋白及其訊息核糖核酸之表現量與控制組比較，則呈現低度表現。然而，事先給予高壓氧前處理再誘導高海拔壓力時，則可有效改善肺水腫及恢復第一型、第五型水通道蛋白質及其訊息核糖核酸在肺組織中表現量。反之，如果在高壓氧前處理後給予熱休克蛋白質70抗體，降低了肺組織中熱休克蛋白質70表現量，則將逆轉高壓氧前處理對於高海拔壓力的保護作用。 結論與建議：高壓氧前處理將可以透過熱休克蛋白質70的表現達到預防高海拔誘導的肺水腫。
Part I Objective: Traumatic brain injury (TBI) causes increased release of several mediators from injured and dead cells and elicits microglial activation. Activated microglia change morphology, migrate to injury sites, and release tumor necrosis factor-alpha (TNF-α) and others. In this study we used a controlled fluid percussion injury model of TBI in the rat to determine whether immediate treatment with MLC601, a traditional Chinese medicine, would affect microglial activation and improve recovery. MLC was chosen for this study because it is beneficial in treating stroke patients and it has been approved for clinical trials. Methods and Materials: A controlled fluid percussion injury model of TBI in the rat was used to determine early treatment (1 h post-injury) or late treatment (4 days postinjury) with MLC 601, would affect microglial activation and improve recovery. Rats were randomly divided into three groups: Sham operation group, TBI with vehicle group, TBI with MLC601 treatment group. Rats with induced TBI were treated with a single intraperitoneal injection of MLC 601 (4 mg/kg) 1h after injury, and then with one injection per day for 2 days. Acute neurological and motor deficits were assessed in all rats the day before and 4 days after injury. An immunofluorescence microscopy method was used to count the numbers of the cells colocalized with neuron- and apoptosis-specific markers, and the cells colocalized with microglia- and TNF-α-specific markers, in the contused brain regions 4 days post-injury. An immunohistochemistry methods was used to evaluate both the number and the morphological transformation of microglia in the injured areas. Results: It was found that early treatment with MLC 601 had better effects in reducing TBI-induced cerebral contusion than did the late therapy with MLC 601. Cerebral contusion caused by TBI was associated with neurological motor deficits, brain apoptosis, and activated microglia (e.g., microgliosis, amoeboid microglia, and microglial overexpression of TNF-α), which all were significantly attenuated by MLC 601 therapy. Conclusion and Suggestion: These results suggest that early MLC601 therapy may improve outcomes of TBI in rats by attenuating microgliosis, morphological transformation of microglia, and microglial overexpression of TNF-α. Part II Objective：HBO2P (hyperbaric oxygen preconditioning) induces the overexpression of HSP70 (heat-shock protein 70) and attenuates brain edema in rats during simulated high-altitude exposure. However, it is not known whether the increased lung injury scores and the decreased levels of both aquaporin (AQP1) and AQP5 proteins and messenger RNA (mRNA) expression in the lung caused by HAE can be affected by HSP70-mediated HBO2P in rats. In the present study, we hypothesized that HBO2P would protect against HAE-induced acute lung injury and edema via promoting HSP70 in lungs prior to the onset of HAE. Methods and Materials： Rats were randomly divided into four groups: the non-HBO2P+ non-HAE group, the non-HBO2P+HAE group, the HBO2P+HAE group, and the HBO2P+HSP-70 antibodies (Abs)+HAE group. The HBO2P groups were given 100% O2 at 2.0 absolute atmospheres for 1 hour per day for 5 consecutive days. The HAE groups were exposed to simulated HAE (9.7% O2 at 0.47 absolute atmospheres of 6,000 m) in a hypobaric chamber for 3 days, polyclonal rabbit anti-mouse HSP-70-neutralizing Abs were intravenously injected 24 hours before the HAE experiments. Immediately after returning to normal atmosphere, the rats were overdosed with a general anesthetic, and then their bungs were excised en bloc for both histologic and molecular evaluation and analysis. Results： In non-HBO2P+HAE group, the animals displayed higher scores of alveolar edema, neutrophil infiltration, and hemorrhage compared with those of non-HBO2P+non-HAE controls. In contrast, the levels of both aquaporin (AQP) 1 and AQP 5 proteins and mRNA expression in the lung in the non-HBO2P+HAE group were significantly lower than those of non-HBO2P controls. The increased lung injury scores and the decreased levels of both AQP1 and AQP5 proteins and mRNA expression in the lung caused by HAE was significantly reduced by HBO2P+HAE. Furthermore, HSP70 Abs, in addition to reducing lung HSP70 proteins, significantly attenuated the beneficial effects of HBO2P in HAE. Conclusion and Suggestion： Our results suggest that high-altitude lung edema can be prevented by HSP-70-mediated HBO2P in rats. |
