摘要
目的从体内和体外分别考察丙三醇的局部刺激性和细胞毒性,初步评价丙三醇作为吸入制剂辅料应用的安全性。方法局部刺激性试验:取Wistar大鼠,分为2组。丙三醇组采用气管内雾化给予20%丙三醇,阴性对照组给予同体积0. 9%氯化钠注射液,连续给药5 d。给药结束后取咽喉部、气管、肺进行组织病理学检查,进行支气管肺泡灌洗并检测灌洗液中总蛋白(TP)含量和碱性磷酸酶(ALP)、乳酸脱氢酶(LDH)活性。细胞毒性试验:采用MTT法研究丙三醇对人肺腺癌细胞A549、人支气管上皮细胞16HBE、大鼠气管上皮细胞RTE这3种细胞株的毒性作用,分别计算IC50值。结果局部刺激性试验:组织病理学检查结果显示,阴性对照组出现轻度肺泡内炎症细胞浸润(1/8),丙三醇组出现极轻度到轻度的肺泡内炎症细胞浸润(6/8)和血管周围炎(2/8);支气管肺泡灌洗液检查结果显示,与阴性对照组相比,丙三醇组ALP水平显著升高(P <0. 05)。细胞毒性试验:在体积分数0. 25%～10%内,丙三醇对A549、16HBE及RTE的细胞毒性作用存在量效关系,相关系数r^2值分别为0. 989、0. 981和0. 964,IC_(50)分别为4. 28%、4. 40%和4. 31%。结论气管内雾化给予体积分数20%丙三醇对大鼠肺部具有轻微刺激性;体积分数达到2. 0%时,丙三醇对A549和16HBE细胞有显著的毒性作用,体积分数达到4. 0%时对RTE细胞有显著的毒性作用。体内外试验结果均提示大剂量丙三醇可能会对肺部产生轻刺激性及毒性作用。
OBJECTIVE To study the local irritative response and cytotoxicity of glycerol in vivo and in vitro,and evaluate the safety of glycerol as an excipient in inhaled preparations preliminarily. METHODS Local irritation test: rats were divided into two groups,the glycerol group received 20% glycerol by intratracheal atomization once a day,followed by 5 d,the control group was given the equal volume of 0. 9% sodium chloride solutions in the same way. After administration,the throat,trachea and lung were taken for histopathological examination,and bronchoalveolar lavage was performed in each group,total protein( TP) content( BCA method) and alkaline phosphatase( ALP),lactate dehydrogenase( LDH) activity in the lavage fluid were detected( biochemical instrument). Cytotoxicity test: the toxic effects of glycerol on adenocarcinoma human alveolar epithelial-like cells( A549),human bronchial epithelial cells( 16 HBE) and rat tracheal epithelial cells( RTE) were studied by MTT assay. RESULTS Local irritation test: the control group showed mild infiltration of inflammatory cells in the alveolar( 1/8),glycerol group showed perivascular inflammation( 2/8) and moderate mild to mild infiltration of inflammatory cells in the alveolar( 6/8);bronchoalveolar lavage fluid test showed that,compared with the control group,the level of ALP in the glycerol group was elevated significantly( P < 0. 05). There was a dose-effect relationship between the cytotoxicity on A549,16 HBE and RTE cells with varying volume fraction of glycerol content in the range of 0. 25%-10%,r^2 values were 0. 989,0. 981,0. 964,and IC50 were 4. 28%,4. 40%,4. 31% respectively. CONCLUSION The 20% glycerol has slight irritation to the lung in vivo,and in vitro glycerol has obvious toxic effect on A549 and 16 HBE cells when the volume fraction reached 2. 0%,when it reached 4. 0%,it has significant toxic effect on RTE cells,suggesting that large dose of glycerol may cause mild irritation and toxicity to the lungs.
作者
冯红敏
盛云华
胡玥
王宇
孙杰
唐黎明
FENG Hong-min;SHENG Yun-hua;HU Yue;WANG Yu;SUN Jie;TANG Li-ming(School of Pharmacy,Fudan University,Shanghai 201203,China;Shanghai Institute for Food and Drug Control,Shanghai 201203,China)
出处
《中国药学杂志》
CAS
CSCD
北大核心
2019年第1期42-46,共5页
Chinese Pharmaceutical Journal
关键词
辅料
丙三醇
气管内雾化
局部刺激性
细胞毒性
excipient
glycerol
intratracheal atomization
local irritation
cytotoxicity
