臺灣博碩士論文加值系統

揮發性碳氫氯化物如二氯甲烷、二氯乙烷和三氯甲烷是工業廣泛使用的有機溶劑，常用於金屬及皮革清洗或粘著劑及殺蟲劑之溶劑上，因為這些有機物具揮發性易擴散至周遭環境而造成嚴重的污染。暴露於揮發性碳氫氯化物的工人曾出現咳嗽、呼吸困難及類似氣喘等呼吸道疾病，然而，對相關的呼吸道急性毒害研究並不多，毒理機制的探討更是缺乏。因為揮發性碳氫氯化物進入人體的主要途徑是經由呼吸道吸入，所以本研究利用等長張力測定的方式檢測三種揮發性碳氫氯化物對呼吸道平滑肌收縮的影響及毒性機制，並以免疫酵素分析法檢測暴露碳氫氯化物對上皮細胞媒介物質釋放之影響。
結果發現二氯甲烷（1000 ppm），二氯乙烷（1000 ppm）及三氯甲烷（³100 ppm）可直接引起氣管平滑肌的基礎收縮，在不同的濃度下（100至1000 ppm）會加強乙醯膽鹼、氯化鉀或組織胺所引起的平滑肌收縮作用，但在高濃度（1000 ppm）時產生兩相反應，先引起收縮後舒張。實驗證實三氯甲烷可促使細胞膜上L-型鈣離子通道打開以引起鈣離子內流及促使內質網中鈣離子的釋放，而增加細胞內鈣離子濃度並引起平滑肌收縮。平滑肌在三氯甲烷高濃度下引起舒張作用可能是因為刺激b腎腺性接受器、活化NO-cGMP路徑或促使鉀離子通道的開啟所致。揮發性碳氫氯化物同時也增加上皮細胞分泌前列腺素E2的能力。
綜合以上結果，當呼吸道暴露於揮發性碳氫氯化物後因影響鈣離子的活動而引起氣管收縮或加強刺激物的作用，同時也促進發炎物質的分泌。所以，由此推測暴露揮發性碳氫氯化物引發呼吸道毒性可能與呼吸道平滑肌的過度緊張及上皮細胞發炎物質的分泌有關。

Volatile chlorinated hydrocarbons (VCHs), such as dichloromethane (DCM), dichloroethane (DCE) and trichloromethane (TCM), are widely used in many industrial processes and are also found in many commercial household products. They might belong to the hazardous air pollutants since VCHs exposure induces respiratory complications including edema, cough, wheezing, and airway hyperactivity. However, studies concerning the contribution of airway responsiveness to VCHs-induced respiratory dysfunction are scarce.
Effects of VCHs, such as DCM, DCE and TCM, on the basal and spasmogen-induced contractile responses in piglet tracheal smooth muscle strips were measured by isometric tension methodology. DCM (1000 ppm), DCE (1000 ppm) and TCM (³100 ppm) induced basal smooth muscle contraction. VCHs (100-1000 ppm) also increased the contractile responses of smooth muscle strips precontracted by ACh, histamine and KCl. Interestingly, high concentration of TCM induced relaxant responses of smooth muscle precontracted by ACh and KCl.
The mechanism of TCM-induced contraction may be due to the influx of external Ca2+ and release of Ca2+ from sarcoplasmic reticulum. The relaxation response elicited by TCM in precontracted smooth muscle, may be due to the activation b-adrenergic receptor, NO-cGMP pathway or potassium channel. Moreover, VCHs increased tracheal epithelial release of PGE2. However the release of PGE2 induced by VCHs exposure did not affect the tracheal muscle tone.
The results demonstrate that exposure of airway to VCHs enhances the basal contractile ability of tracheal smooth muscle, potentiates the stimulant-induced responses and induces the epithelial PGE2 release, suggesting that airway dysfunction induced by inhaled VCHs may be due to the hyperresponsiveness of airway smooth muscle.

目 錄
頁次
英文摘要…………………………………………………………………………1
中文摘要…………………………………………………………………………2
緒言………………………………………………………………………………3
揮發性碳氫氯化合物………………………………………………………3
呼吸道的調節………………………………………………………………7
研究假說………………………………………………………………………..12
研究目的………………………………………………………………………..13
方法與材料……………………………………………………………………..14
實驗結果………………………………………………………………………..17
揮發性碳氫氯化物對氣管平滑肌張力之影響……………………….. ...17
三氯甲烷引起氣管平滑肌收縮的機制…………………………………..18
三氯甲烷舒張乙醯膽鹼引起氣管平滑肌張力的機制………………..20
揮發性碳氫氯化物對氣管上皮細胞分泌能力的影響………………………..22
綜合比較揮發性碳氫氯化物對氣管上皮細胞及平滑肌張力之影響…..22
討論……………………………………………………………………………..23
結論……………………………………………………………………………..29
參考文獻…………………………………………………………………….….30
圖表……………………………………………………………………………..37
圖表目錄
頁
Table 1 Basal contractile responses induced by VCHs in piglet tracheal smooth
muscle strips ………….……………………………………………..……….37
Table 2 Effects of atropine, prazosin and phentolamine on TCM-induced contraction………………..……………….…………………………….…...38
Table 3 Effects of nifedipine and verapamil on TCM-induced contraction……….….39
Table 4 The smooth muscle tension induced by acetylcholine and carbachol with
the presence of epithelium…………………………………………….....……40
Figure 1 Representative effects of VCHs on the isometric tension of piglet
tracheal smooth muscle strips………………...………………….…………..41
Figure 2 Concentration-response curves for DCM on ACh-induced muscle tension...42
Figure 3 Concentration-response curves for DCE on ACh-induced muscle tension....43
Figure 4 Concentration-response curves for TCM on ACh-induced muscle tension...44
Figure 5 Concentration-response curves for DCM on KCl-induced muscle tension....45
Figure 6 Concentration-response curves for DCE on KCl-induced muscle tension...46
Figure 7 Concentration-response curves for TCM on KCl-induced muscle tension....47
Figure 8 Concentration-response curves for VCHs on histamine-induced muscle tension…………………………….….……………..……………….……….48
Figure 9 Effects of pretreatment with VCHs on smooth muscle contraction induced
by spasmogens……………………………………………………………..49
Figure 10 TCM induced contraction in normal or Ca2+-free external solution...……50
Figure 11 Effects of caffeine and ryanodine on TCM-induced contraction………….51
Figure 12 TCM attenuated the relaxant effects of isoproterenol………….…………..53
Figure 13 Effects of propranolol on TCM-induced relaxation……………………….54
頁
Figure 14 TCM attenuated the relaxant effects of forskolin ……………………...….56
Figure 15 TCM attenuated the relaxant effects of sodium nitroprusside……………..57
Figure 16 Effects of methylene blue on TCM-induced relaxation………………….58
Figure 17 Effects of TEA on TCM-induced relaxation……………………...………..59
Figure 18 Effects of glibenclamide on TCM-induced relaxation…………………..61
Figure 19 Effects VCHs on PGE2 release in tracheal epithelium…………………..63
Figure 20 Effects of VCHs on the activity of acetylcholinesterase in tracheal epithelium………..………………………………………………………...64
Figure 21 Effects of acetylcholine and carbachol-induced smooth muscle contraction in the absence or presence of epithelium………...………………………...65

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