荖葉萃取物與檳榔萃取物誘發小鼠心血管反應之比較應用

Pharmacological investigation of Piper betle extracts and Betel quid extracts for behavior in mouse

林羿漢(Yi-Han Lin)

指導教授 ： 翁清松 ； 王偉傑

中原大學/工學院/生物醫學工程研究所/碩士(2017年)

https://doi.org/10.6840/cycu201700386

摘要

口腔癌是台灣人癌症十大死因之一，在臨床發現每10位口腔癌患者，有9位有嚼食檳榔的惡習，台灣口腔癌發生率更是世界之冠。國際癌症研究總署也證實，檳榔子為第一類致癌物。研究顯示，咀嚼檳榔容易誘發多種系統相關疾病，包含心血管系統與心率不整等相關疾病。後代也有高達2倍的風險發生代謝症候群，但縱使再多文獻研究清楚訴說咀嚼檳榔的危害，但仍無法改善國人咀嚼者的習慣。

本研究的目的為尋找漸進式替代咀嚼檳榔的無害物，以降低食用檳榔者造成心率不整為目的，故查閱文獻後探討檳榔添加的附屬品中的荖葉，其是否能造成心血管神經系統產生反應，進一步達到影響食用者影響神經系統的相聯性反應。

本研究的第一步透過不同萃取溶劑(水與乙醇)各別萃取檳榔與荖葉將萃取溶液進行減壓濃縮提高有效成份含量。第二步為分析化驗萃取液各別所含有效影響因子的成份量，選擇萃取溶液有效成份較高的乙醇萃取物並計算因施予的劑量。第三步為動物實驗，以90%生理食鹽水(PBS)與10%乙醇為對照組，比對檳榔萃取液與荖葉萃取液於小鼠(ICR)的腹腔取檳榔鹼與丁香酚各別為5mg/1kg的比例來注射，並觀測小鼠的EKG反應與記錄；再以同劑量於一周後施打於小鼠腹腔，進行曠野測試，透過小鼠活動量與活動模式變化，分析評估其運動能力、焦慮程度等變化。

研究結果，於萃取溶劑的選擇上透過GC化驗，確認不論乙醇與水都能萃取出預期的神經影響成份，故未來是能進行相關產品的研製；而檳榔與荖葉萃取液進行的動物實驗來探討預期的影響成份是否真有效用，在EKG的測試中，潘凱圖觀測到檳榔萃取液在施予5mg/1kg的比例時已經造成小鼠產生檳榔中毒的跡象而自律神經整體反應上是大於荖葉萃取液的，但荖葉萃取液依然對自律神經有所影響。心電圖數據上荖葉萃取液有明顯與副交感神經產生反應；檳榔萃取液則是交感與副交感神經處於極度不穩定的情況。

小鼠曠野測試上則藉由進入中央區域與總行徑路程分別判斷小鼠的焦慮感及亢奮程度，在檳榔萃取液因已深切影響小鼠的自律神經，故小鼠呈現強烈的不適感，在進入中央區域與總行徑路程都適極低的反應；荖葉萃取液無論在進入中央區域的次數或是總行徑路程是大過於對照組的數據，表示在生物物理性上是具有解焦慮與提神亢奮的效果，透過本研究的荖葉萃取液的成份分析、心電測試與動物實驗能充分了解荖葉應用於取代檳榔的實際方式與成效。

關鍵字

荖葉 ； 檳榔

並列摘要

Oral cancer is one of the top ten leading causes of death in Taiwan. Clinical study shows that approximately 9 people in every 10 people with oral cancer in clinic have the habit of betel quid (BQ) chewing. The incidence rate of oral cancer in Taiwan is the highest in the world. International Agency for Research on Cancer (IARC) considered arecoline as a group 1 carcinogen.

Researches show that BQ chewing can induce a variety of system-related diseases and double the risk of metabolic syndrome in the next generation. Although many studies reveal the negative effects of BQ chewing, it is still hard to reduce the number of people having this habit.

The purpose of this study was to find a harmless substance to replace the BQ gradually. We used the non-carcinogenic piper betel to study whether it will influence the cardiovascular and nervous system, and to investigate if eating the piper betel will affect the reaction of the nerve system.

The first step of this study was to increase the content of active ingredient by water and ethanol extraction from the betel quid and piper betel. The second step was to analyze the content of effector in each extract. The extract with higher content of active ingredient was chosen and the dosage level was calculated. The third step was to conduct animal testing. ICR mice were intraperitoneally injected with 5 mg/kg arecoline, which was extracted from BQ, or 5 mg/kg Eugenol, which was extracted from piper betel). Control group was treated with 90% PBS and 10% ethanol. The EKG reaction of mice was observed and recorded. After one week, mice were intraperitoneally injected with the same dosage. Then, the Open field test (OFT) was conducted. The activity amount and activity model were observed to analyze the change of the activity ability and anxiety in mice.

The results showed that the expected active ingredient can be extracted by both ethanol and water. Therefore, the relevant products could be developed in the future. In the EKG test, the Poincaré plot indicated that the mice treated with 5 mg/kg arecoline (BQ extract) were poisoned. Additionally, the response of autonomic nerve of BQ extract was more active than the piper betel extract. However, the piper betel extract still has an impact on the autonomic nerve system. The data of EKG showed piper betel extract had a significant effect on bradycardia, while the BQ extract caused the extremely unstable situation.

The degree of the anxiety and excitement were determined by how the mice entered the central area and the total route in the OFT. Due to the strong impact on autonomic nerve, the mice treated with BQ extract suffered great discomfort and showed extremely low reaction when entering the central area and the total route. Compared with the control group, the mice treated with piper betel showed more times of entering central area and total route. It revealed that the piper betel can reduce the anxiety, feel refreshed and feel excited. This study showed the practical way and the effort of the application of piper betel to replace BQ by analyzing the content of piper betel extract, and conducting the EKG and animal test.

並列關鍵字

Piper betle ； Betel quid

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