中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2012年
32期
2283-2287
,共5页
李敏超%尤列·皮尔曼%维克多·科罗索夫%周向东
李敏超%尤列·皮爾曼%維剋多·科囉索伕%週嚮東
리민초%우렬·피이만%유극다·과라색부%주향동
瞬时受体电位通道%黏蛋白类%大鼠
瞬時受體電位通道%黏蛋白類%大鼠
순시수체전위통도%점단백류%대서
Transient receptor potential channels%Mucins%Rat
目的 探讨冷空气联合香烟烟雾吸入诱导大鼠气道黏液高分泌的机制及雪莲、布地奈德对该过程的干预作用.方法 SD大鼠70只被随机分为7组:A组为对照组；B组为冷空气刺激组(吸入-18 ℃冷空气,3 h/d,持续40 d)；C组为香烟烟雾吸入组(吸入香烟烟雾,0.5 h/d,持续40 d)；D组为冷空气刺激+香烟烟雾吸入组；E组为冷空气刺激+香烟烟雾吸入+雪莲干预组(腹腔内注入雪莲注射液0.8 mg/kg,1次/d,持续40 d)；F组为冷空气刺激+香烟烟雾吸入+布地奈德干预组(吸入布地奈德,0.5 mg/kg,1次/d,持续40 d)；G组为冷空气刺激+香烟烟雾吸入+雪莲+布地奈德干预组(药物用法同前).实时荧光定量PCR法检测各组大鼠支气管上皮内TRPM8 mRNA相对水平,免疫组化法、Western印迹法检测各组大鼠支气管上皮内TRPM8蛋白相对水平,酶联免疫吸附法检测支气管肺泡灌洗液(BALF)中黏蛋白(MUC)5AC、白细胞介素8(IL-8)和肿瘤坏死因子α(TNF-α.)表达.结果 A～G组支气管上皮内TRPM8 mRNA相对表达量分别为1.00±0.00、0.98±0.07、2.27±0.29、2.31 ±0.30、1.55±0.38、1.66±0.40、1.31±0.23；免疫组化法和Western印迹法检测TRPM8蛋白相对表达量结果分别为0.16 ±.0.05、0.16 ±0.04、0.22 ±0.06、0.25 ±0.05、0.17±0.04、0.18±0.03、0.15±0.05和0.25 ±0.04、0.24 ±0.03、0.58 ±0.06、0.56 ±0.09、0.41 ±0.09、0.39±0.07、0.20 ±0.06;C、D组均显著高于A组,E、F、G组均显著低于D组(均P＜0.05).A～G组BALF中MUC5AC水平分别为(57±6)、(69±5)、(66±4)、(185 ±43)、(142±30)、(147±36)、(60±11)μg/mg,IL-8水平分别为(58±14)、(146±38)、(134±29)、(379±101)、(262 ±67)、(294 ±70)、(81±27) ng/L,TNF-α水平分别为(153±28)、(208±90)、(274±64)、(600±113)、(458±96)、(498 ±84)、(169 ±65) ng/L;B、C、D组均显著高于A组,E、F、G组均显著低于D组(均P＜0.05),且香烟烟雾吸入与冷空气刺激、雪莲与布地奈德具有协同作用.结论 冷空气刺激通过激活由香烟烟雾吸入所诱导的高水平TRPM8通道使前炎性细胞因子及MUC5AC呈现过度释放状态；雪莲、布地奈德对该过程具有协同性抑制作用.
目的 探討冷空氣聯閤香煙煙霧吸入誘導大鼠氣道黏液高分泌的機製及雪蓮、佈地奈德對該過程的榦預作用.方法 SD大鼠70隻被隨機分為7組:A組為對照組；B組為冷空氣刺激組(吸入-18 ℃冷空氣,3 h/d,持續40 d)；C組為香煙煙霧吸入組(吸入香煙煙霧,0.5 h/d,持續40 d)；D組為冷空氣刺激+香煙煙霧吸入組；E組為冷空氣刺激+香煙煙霧吸入+雪蓮榦預組(腹腔內註入雪蓮註射液0.8 mg/kg,1次/d,持續40 d)；F組為冷空氣刺激+香煙煙霧吸入+佈地奈德榦預組(吸入佈地奈德,0.5 mg/kg,1次/d,持續40 d)；G組為冷空氣刺激+香煙煙霧吸入+雪蓮+佈地奈德榦預組(藥物用法同前).實時熒光定量PCR法檢測各組大鼠支氣管上皮內TRPM8 mRNA相對水平,免疫組化法、Western印跡法檢測各組大鼠支氣管上皮內TRPM8蛋白相對水平,酶聯免疫吸附法檢測支氣管肺泡灌洗液(BALF)中黏蛋白(MUC)5AC、白細胞介素8(IL-8)和腫瘤壞死因子α(TNF-α.)錶達.結果 A～G組支氣管上皮內TRPM8 mRNA相對錶達量分彆為1.00±0.00、0.98±0.07、2.27±0.29、2.31 ±0.30、1.55±0.38、1.66±0.40、1.31±0.23；免疫組化法和Western印跡法檢測TRPM8蛋白相對錶達量結果分彆為0.16 ±.0.05、0.16 ±0.04、0.22 ±0.06、0.25 ±0.05、0.17±0.04、0.18±0.03、0.15±0.05和0.25 ±0.04、0.24 ±0.03、0.58 ±0.06、0.56 ±0.09、0.41 ±0.09、0.39±0.07、0.20 ±0.06;C、D組均顯著高于A組,E、F、G組均顯著低于D組(均P＜0.05).A～G組BALF中MUC5AC水平分彆為(57±6)、(69±5)、(66±4)、(185 ±43)、(142±30)、(147±36)、(60±11)μg/mg,IL-8水平分彆為(58±14)、(146±38)、(134±29)、(379±101)、(262 ±67)、(294 ±70)、(81±27) ng/L,TNF-α水平分彆為(153±28)、(208±90)、(274±64)、(600±113)、(458±96)、(498 ±84)、(169 ±65) ng/L;B、C、D組均顯著高于A組,E、F、G組均顯著低于D組(均P＜0.05),且香煙煙霧吸入與冷空氣刺激、雪蓮與佈地奈德具有協同作用.結論 冷空氣刺激通過激活由香煙煙霧吸入所誘導的高水平TRPM8通道使前炎性細胞因子及MUC5AC呈現過度釋放狀態；雪蓮、佈地奈德對該過程具有協同性抑製作用.
목적 탐토랭공기연합향연연무흡입유도대서기도점액고분비적궤제급설련、포지내덕대해과정적간예작용.방법 SD대서70지피수궤분위7조:A조위대조조；B조위랭공기자격조(흡입-18 ℃랭공기,3 h/d,지속40 d)；C조위향연연무흡입조(흡입향연연무,0.5 h/d,지속40 d)；D조위랭공기자격+향연연무흡입조；E조위랭공기자격+향연연무흡입+설련간예조(복강내주입설련주사액0.8 mg/kg,1차/d,지속40 d)；F조위랭공기자격+향연연무흡입+포지내덕간예조(흡입포지내덕,0.5 mg/kg,1차/d,지속40 d)；G조위랭공기자격+향연연무흡입+설련+포지내덕간예조(약물용법동전).실시형광정량PCR법검측각조대서지기관상피내TRPM8 mRNA상대수평,면역조화법、Western인적법검측각조대서지기관상피내TRPM8단백상대수평,매련면역흡부법검측지기관폐포관세액(BALF)중점단백(MUC)5AC、백세포개소8(IL-8)화종류배사인자α(TNF-α.)표체.결과 A～G조지기관상피내TRPM8 mRNA상대표체량분별위1.00±0.00、0.98±0.07、2.27±0.29、2.31 ±0.30、1.55±0.38、1.66±0.40、1.31±0.23；면역조화법화Western인적법검측TRPM8단백상대표체량결과분별위0.16 ±.0.05、0.16 ±0.04、0.22 ±0.06、0.25 ±0.05、0.17±0.04、0.18±0.03、0.15±0.05화0.25 ±0.04、0.24 ±0.03、0.58 ±0.06、0.56 ±0.09、0.41 ±0.09、0.39±0.07、0.20 ±0.06;C、D조균현저고우A조,E、F、G조균현저저우D조(균P＜0.05).A～G조BALF중MUC5AC수평분별위(57±6)、(69±5)、(66±4)、(185 ±43)、(142±30)、(147±36)、(60±11)μg/mg,IL-8수평분별위(58±14)、(146±38)、(134±29)、(379±101)、(262 ±67)、(294 ±70)、(81±27) ng/L,TNF-α수평분별위(153±28)、(208±90)、(274±64)、(600±113)、(458±96)、(498 ±84)、(169 ±65) ng/L;B、C、D조균현저고우A조,E、F、G조균현저저우D조(균P＜0.05),차향연연무흡입여랭공기자격、설련여포지내덕구유협동작용.결론 랭공기자격통과격활유향연연무흡입소유도적고수평TRPM8통도사전염성세포인자급MUC5AC정현과도석방상태；설련、포지내덕대해과정구유협동성억제작용.
Objective To explore the mechanisms of mucus hypersecretion in airway of rats induced by the synergies between cold air and cigarette smoke inhalation and understand the intervention effects of saussurea and budesonide in this process. Methods A total of 70 SD rats were randomly divided into 7 groups.Group A:control; Group B:cold stimulation group receiving cold air inhalation for 3 h daily for 40 d; Group C:cigarette smoke inhalation group receiving cigarette smoke inhalation for 0.5h daily for 40 d;Group D:cigarette smoke inhalation + cold stimulation group; Group E:cigarette smoke inhalation + cold stimulation + saussurea (0.8 mg/kg saussurea intraperitoneally injected once daily for 40 d) ; Group F:cigarette smoke inhalation + cold stimulation + inhaled budesonide (0.5 mg/kg inhaled once daily for 40 d) ; Group G:cigarette smoke inhalation + cold stimulation + saussurea + inhaled budesonide.The relative quantities of TRPM8 mRNA within bronchial epithelium of each group were detected by real-time polymerase chain reaction (PCR) and TRPM8 protein was detected by immunohistochemical assay and Western blot.The levels of mucin (MUC) 5AC,interleukin 8 (IL-8) and tumor necrosis factor alpha (TNF-α) in bronchoalveolar lavage fluid (BALF) were detected by enzyme-linked immunosorbent assay.Results TRPM8 mRNA of groups A-G were 1.00 ±0.00,0.98 ±0.07,2.27 ±0.29,2.31 ±0.30,1.55 ±0.38,1.66 ± 0.40 and 1.31 ± 0.23 ; TRPM8 protein 0.16 ± 0.05,0.16 ± 0.04,0.22 ± 0.06,0.25 ±0.05,0.17 ±0.04,0.18 ±0.03,0.15 ±0.05,0.25 ±0.04,0.24 ±0.03,0.58 ±0.06,0.56 ±0.09,0.41 ±0.09,0.39 ±0.07 and 0.20 ±0.06 respectively.TRPM8 mRNA and protein of groups C and D were significantly higher than those of group A.And groups E,F and G were significantly lower than those of group D (allP＜0.05).In BALF of groups A-G,MUC5AC were (57±6),(69±5),(66±4),(185 ±43),(142 ±30),(147 ±36) and (60 ±11) μg/mg,IL-8 (58 ±14),(146 ±38),(134 ±29),(379 ±101),(262 ±67),(294 ±70) and (81 ±27) ng/L,TNF-α(153 ±28),(208 ±90),(274 ±64),(600 ± 113),(458 ±96),(498 ±84) and (169 ±65) ng/L respectively.The values of groups B,C and D were significantly higher than those of group A ( all P ＜ 0.05 ) while groups E,F and G were significantly lower than those of group D ( all P ＜ 0.05).Cigarette smoke inhalation and cold stimulation synergistically enhanced the expression of MUC5AC,IL-8 and TNF-α. Saussurea and inhaled budesonide synergistically inhibited the expression of MUC5AC,IL-8 and TNF-α. Conclusions Cold air inhalation evokes the release of proinflammatory cytokines and MUC5AC via activated TRPM8 channel up-regulated by cigarette smoke inhalation.Saussurea and inhaled budesonide synergistically inhibits the above mentioned process.
