二乙二醇類農藥其他成分對小鼠免疫細胞與特異性抗體之影響

Effects of Inert Diethylene Glycols in Pesticides on Immunocytes and Specific Antibodies in Mice

吳偉嘉(Wei-Jia Wu) ； 廖俊麟(Chun-Lin Liao) ； 蔡韙任(Wei-Ren Tsai)

《臺灣農藥科學》 2期 (2017/06) Pp. 55-74

https://doi.org/10.6671/TPS/2017.002.05

摘要

本研究以小鼠免疫毒性測試方法， 評估國內外常用二乙二醇類 (diethylene glycols) 農藥其他成分 (inert ingredients) 對小鼠免疫系統之影響。試驗分為體外 (in vitro) 與體內 (in vivo) 試驗。In vitro 試驗為評估二乙二醇類農藥其他成分對小鼠脾臟淋巴細胞之影響，並以此細胞增殖模式評估3 種二乙二醇類農藥其他成分，其試驗濃度分別為0、2.5 mM、5 mM、10 mM 及25 mM。結果顯示，正對照組藥劑環磷醯胺(cyclophosphamide, CPA)、二乙醇單丁醚 (diethylene glycol monobutyl ether, DGBE)於濃度 (2.5 ~ 25 mM) 可明顯抑制脾臟T 與B 淋巴細胞的增殖作用 (P ＜ 0.05)；二乙醇單甲醚 (diethylene glycol monomethyl ether, DGME) 於濃度 (2.5 ~ 25 mM) 可明顯抑制脾臟T 淋巴細胞的增殖作用 (P ＜ 0.05)；二乙醇單乙醚 (diethylene glycol monoethyl ether, DGEE) 對小鼠之脾臟淋巴細胞的增殖則無顯著差異。進一步以明顯抑制脾臟淋巴細胞的DGBE 進行in vivo 試驗，以胃管投予DGBE (50、100 及200 mg/kg) 進行28 d 之免疫毒性，並以CPA 為正對照組藥劑，評估其對BALB/c 品系小鼠之免疫反應。分析項目包括脾臟活性細胞數 (cellularity) 與淋巴細胞增殖反應、血液相、免疫球蛋白M (immunoglobulin M, IgM)、脾臟淋巴細胞分型及組織病理變化。結果顯示，DGBE 處理組對小鼠之體重、臟器重、紅血球總數、白血球總數、血球容積比及血色素等與空白對照組相較均無顯著差異，CPA 處理組對小鼠之胸腺重量呈現明顯的降低(P ＜ 0.05)。白血球分類計數 (WBC differential count) 結果顯示，DGBE 處理組對小鼠之淋巴球、單核球、嗜中性球、嗜伊紅性球及嗜鹼性球之比例上亦無顯著差異，CPA處理組小鼠血液中淋巴球比例與空白對照組相較呈現明顯的減少現象 (P ＜ 0.05)。脾臟活性細胞總數與淋巴細胞增殖試驗結果顯示，DGBE 處理組小鼠之脾臟活性細胞總數與其T、B 淋巴細胞in vivo 增殖均無顯著差異；CPA 處理組小鼠之脾臟活性細胞總數呈現明顯的抑制效應 (P ＜ 0.05)，但其T、B 淋巴細胞in vivo 增殖數量呈現顯著的上升 (P ＜ 0.05)。DGBE 處理組對小鼠之IgM 抗體與淋巴細胞分型均無顯著差異，CPA 處理組對小鼠之IgM 抗體與B 淋巴細胞比例具明顯的抑制 (P ＜ 0.05)。組織病理觀察結果顯示，DGBE 處理組均無明顯組織病理變化，CPA 處理組可見小鼠胸腺細胞明顯減少 (P ＜ 0.05)，而呈現胸腺組織細胞排列鬆散情形。綜合以上結果顯示，DGBE 對小鼠在此試驗下無明顯的免疫毒性，其28 d 免疫毒性之無可見毒害劑量 (no observed adverse effect level, NOAEL) 為200 mg/kg/day。

關鍵字

二乙二醇類 ； 農藥其他成分 ； 免疫毒性 ； 無可見毒害劑量

並列摘要

In this study, both in vitro and in vivo tests were performed to elucidate the effects of diethylene glygols, which comprise inert ingredients in pesticides, on immunocytes and specific antibodies in mice. For the in vitro test, the proliferation of mouse splenocyte cells was quantified to determine the effects of DGME (diethylene glycol monomethyl ether), DGEE (diethylene glycol monoethyl ether) and DGBE (diethylene glycol monobutyl ether), each of which was administered at 5 different concentrations (0, 2.5, 5, 10, and 25 mM). We found that only DGBE significantly influenced the proliferation of spleen lymphocytes at all treatment levels. Consequently, a 28-day oral immunotoxicity test was performed, in which mice were administered 50, 100, or 200 mg/kg DGBE. (For this, cyclophosphamide (CPA) was used as a positive control.) Assay results did not show significant differences in terms of of spleen cellularity, lymphocyte proliferation, IgM antibodies, various spleen lymphocytes, hematological data, or histopathological data between the DGBE-treatment group and the blank control group. Conversely, as we expected, CPA was found to significantly reduce thymus weight, spleen cellularity, IgM antibodies, and lymphocyte counts in mice. Taken together, the no observed adverse effect level (NOAEL) of DGBE of this 28-days immunotoxicity test was estimated to be 200 mg/kg/day in mice.

並列關鍵字

diethylene glycols ； inert pesticide ingredients ； immunotoxicity ； no observed adverse effect level