中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
32期
2535-2539
,共5页
哮喘%木犀草素%过氧化物酶体增殖物激活受体%p38丝裂原活化蛋白激酶类%大鼠
哮喘%木犀草素%過氧化物酶體增殖物激活受體%p38絲裂原活化蛋白激酶類%大鼠
효천%목서초소%과양화물매체증식물격활수체%p38사렬원활화단백격매류%대서
Asthma%Luteolin%Peroxisome proliferator-activated raceptors%p38 Mitogen-activated protein kinases%Rats
目的：探讨木犀草素对哮喘大鼠气道炎症的影响机制。方法 SPF级雄性SD大鼠48只，用随机数字表法随机平均分为对照组、哮喘组、木犀草素组3组：哮喘组、木犀草素组复制大鼠哮喘模型，即在第1、8天腹腔注射卵白蛋白/氢氧化铝混合液，2周后以1％的卵白蛋白生理盐水溶液雾化激发，每周3次，持续8周。对照组参照上述方法，但以生理盐水代替卵白蛋白/氢氧化铝混合液及卵白蛋白生理盐水。每次激发后30 min，对照组与哮喘组予生理盐水腹腔注射；木犀草素组予1 mg/kg的木犀草素腹腔注射。光镜观察肺组织病理变化，检测支气管肺泡灌洗液（ BALF）中细胞数和白细胞介素4（IL-4）水平，免疫组化法检测各组过氧化物酶体增殖物激活受体γ（PPARγ）蛋白和p38丝裂原活化蛋白激酶（ p38MAPK）蛋白相对表达量；逆转录（ RT）-PCR分别检测PPARγmRNA、p38MAPK mRNA的相对表达量。结果哮喘组大鼠支气管管壁厚度、平滑肌厚度分别为（93.3±7.4）、（34.9±2.3）μm，均显著高于对照组的（61.9±8.2）、（19.3±1.5）μm及木犀草素组的（76.6±6.7）、（25.4±4.6）μm（均P＜0.05）；哮喘组BALF中细胞总数、中性粒细胞数、嗜酸粒细胞数和IL-4水平分别为（5.61±0.63）×109/L、（1.83±0.09）×109/L、（0.59±0.09）×109/L和（78.23±12.73） pg/ml，均显著高于对照组的（1.53±0.31）×109/L、（0.45±0.21）×109/L、（0.07±0.03）×109/L和（21.21±2.53）pg/ml（均P＜0.01）及木犀草素组的（3.24±0.25）×109/L、（1.54±0.10）×109/L、（0.33±0.05）×109/L和（43.24±8.65）pg/ml（均P＜0.05）；哮喘组p38MAPK蛋白相对表达量均显著高于对照组、木犀草素组（0.362±0.008比0.143±0.017、0.251±0.021，均P＜0.01）；对照组、木犀草素组PPARγ蛋白相对表达量均显著高于哮喘组（0.331±0.056、0.442±0.031比0.247±0.034，均P＜0.05）；对照组、木犀草素组p38MAPK mRNA相对表达量均显著低于哮喘组（0.312±0.052、0.426±0.067比0.718±0.064，均P＜0.01）；对照组、木犀草素组PPARγmRNA相对表达量均显著高于哮喘组（0.573±0.042、0.687±0.054比0.266±0.036，均P＜0.01）。结论木犀草素可能是通过影响PPARγ表达及p38MAPK信号通路，抑制哮喘大鼠气道炎症的作用。
目的：探討木犀草素對哮喘大鼠氣道炎癥的影響機製。方法 SPF級雄性SD大鼠48隻，用隨機數字錶法隨機平均分為對照組、哮喘組、木犀草素組3組：哮喘組、木犀草素組複製大鼠哮喘模型，即在第1、8天腹腔註射卵白蛋白/氫氧化鋁混閤液，2週後以1％的卵白蛋白生理鹽水溶液霧化激髮，每週3次，持續8週。對照組參照上述方法，但以生理鹽水代替卵白蛋白/氫氧化鋁混閤液及卵白蛋白生理鹽水。每次激髮後30 min，對照組與哮喘組予生理鹽水腹腔註射；木犀草素組予1 mg/kg的木犀草素腹腔註射。光鏡觀察肺組織病理變化，檢測支氣管肺泡灌洗液（ BALF）中細胞數和白細胞介素4（IL-4）水平，免疫組化法檢測各組過氧化物酶體增殖物激活受體γ（PPARγ）蛋白和p38絲裂原活化蛋白激酶（ p38MAPK）蛋白相對錶達量；逆轉錄（ RT）-PCR分彆檢測PPARγmRNA、p38MAPK mRNA的相對錶達量。結果哮喘組大鼠支氣管管壁厚度、平滑肌厚度分彆為（93.3±7.4）、（34.9±2.3）μm，均顯著高于對照組的（61.9±8.2）、（19.3±1.5）μm及木犀草素組的（76.6±6.7）、（25.4±4.6）μm（均P＜0.05）；哮喘組BALF中細胞總數、中性粒細胞數、嗜痠粒細胞數和IL-4水平分彆為（5.61±0.63）×109/L、（1.83±0.09）×109/L、（0.59±0.09）×109/L和（78.23±12.73） pg/ml，均顯著高于對照組的（1.53±0.31）×109/L、（0.45±0.21）×109/L、（0.07±0.03）×109/L和（21.21±2.53）pg/ml（均P＜0.01）及木犀草素組的（3.24±0.25）×109/L、（1.54±0.10）×109/L、（0.33±0.05）×109/L和（43.24±8.65）pg/ml（均P＜0.05）；哮喘組p38MAPK蛋白相對錶達量均顯著高于對照組、木犀草素組（0.362±0.008比0.143±0.017、0.251±0.021，均P＜0.01）；對照組、木犀草素組PPARγ蛋白相對錶達量均顯著高于哮喘組（0.331±0.056、0.442±0.031比0.247±0.034，均P＜0.05）；對照組、木犀草素組p38MAPK mRNA相對錶達量均顯著低于哮喘組（0.312±0.052、0.426±0.067比0.718±0.064，均P＜0.01）；對照組、木犀草素組PPARγmRNA相對錶達量均顯著高于哮喘組（0.573±0.042、0.687±0.054比0.266±0.036，均P＜0.01）。結論木犀草素可能是通過影響PPARγ錶達及p38MAPK信號通路，抑製哮喘大鼠氣道炎癥的作用。
목적：탐토목서초소대효천대서기도염증적영향궤제。방법 SPF급웅성SD대서48지，용수궤수자표법수궤평균분위대조조、효천조、목서초소조3조：효천조、목서초소조복제대서효천모형，즉재제1、8천복강주사란백단백/경양화려혼합액，2주후이1％적란백단백생리염수용액무화격발，매주3차，지속8주。대조조삼조상술방법，단이생리염수대체란백단백/경양화려혼합액급란백단백생리염수。매차격발후30 min，대조조여효천조여생리염수복강주사；목서초소조여1 mg/kg적목서초소복강주사。광경관찰폐조직병리변화，검측지기관폐포관세액（ BALF）중세포수화백세포개소4（IL-4）수평，면역조화법검측각조과양화물매체증식물격활수체γ（PPARγ）단백화p38사렬원활화단백격매（ p38MAPK）단백상대표체량；역전록（ RT）-PCR분별검측PPARγmRNA、p38MAPK mRNA적상대표체량。결과효천조대서지기관관벽후도、평활기후도분별위（93.3±7.4）、（34.9±2.3）μm，균현저고우대조조적（61.9±8.2）、（19.3±1.5）μm급목서초소조적（76.6±6.7）、（25.4±4.6）μm（균P＜0.05）；효천조BALF중세포총수、중성립세포수、기산립세포수화IL-4수평분별위（5.61±0.63）×109/L、（1.83±0.09）×109/L、（0.59±0.09）×109/L화（78.23±12.73） pg/ml，균현저고우대조조적（1.53±0.31）×109/L、（0.45±0.21）×109/L、（0.07±0.03）×109/L화（21.21±2.53）pg/ml（균P＜0.01）급목서초소조적（3.24±0.25）×109/L、（1.54±0.10）×109/L、（0.33±0.05）×109/L화（43.24±8.65）pg/ml（균P＜0.05）；효천조p38MAPK단백상대표체량균현저고우대조조、목서초소조（0.362±0.008비0.143±0.017、0.251±0.021，균P＜0.01）；대조조、목서초소조PPARγ단백상대표체량균현저고우효천조（0.331±0.056、0.442±0.031비0.247±0.034，균P＜0.05）；대조조、목서초소조p38MAPK mRNA상대표체량균현저저우효천조（0.312±0.052、0.426±0.067비0.718±0.064，균P＜0.01）；대조조、목서초소조PPARγmRNA상대표체량균현저고우효천조（0.573±0.042、0.687±0.054비0.266±0.036，균P＜0.01）。결론목서초소가능시통과영향PPARγ표체급p38MAPK신호통로，억제효천대서기도염증적작용。
Objective To explore the regulatory effects of luteolin on airway inflammation in asthmatic rats.Methods A total of 48 male Sprague-Dawley ( SD) rats were randomly divided into 3 groups of control , asthmatic and luteolin ( n =16 each ).The rat model of bronchial asthma was established in asthmatic and luteolin groups.The model was induced by intraperitoneally injecting a mixture of ovalbumin and aluminum hydroxide at Day 1 and 8.After two weeks , aomization excitation of normal saline ( containing 1%ovalbumin ) was induced thrice weekly.The treatment lasted 8 weeks.In control group , the mixture of ovalbumin , aluminum hydroxide and normal saline containing 1%ovalbumin was replaced by normal saline.At 30 min after aomization excitation , normal saline was given to rats in control and asthmatic groups , while 1 mg/kg luteolin was given intraperitoneally to luteolin group.The inflammatory cell number and level of interleukin-4 ( IL-4) were measured in bronchoalveolar lavage fluid ( BALF).The histopathological changes were observed under light microscope.The activities of peroxisome proliferator-activated receptors ( PPARγ) and p38 mitogen-activated protein kinases ( p38MAPK ) in pulmonary tissues were detected by immunohistochemistry and reverse transcription-polymerase chain reaction ( RT-PCR ) .Results The bronchial wall thickness of asthma group , along with smooth muscle thickness ( ( 93.3 ±7.4 ) , ( 34.9 ± 2.3) μm) was more than that of control ((61.9 ±8.2), (19.3 ±1.5) μm) and luteolin ((76.6 ±6.7), (25.4 ±4.6) μm) groups (all P<0.05).The total cell count ((5.61 ±0.63) ×109/L), neutrophil count ((1.83 ±0.09) ×109/L), eosinophil count ((0.59 ±0.09) ×109/L) and level of IL-4 ((78.23 ± 12.73) pg/ml) in BALF of asthmatic group were markedly higher than those of control ((1.53 ±0.31) × 109/L, (0.45 ±0.21) ×109/L,(0.07 ±0.03) ×109/L and (21.21 ±2.53) pg/ml) and luteolin ((3.24 ±0.25) ×109/L, (1.54 ±0.10) ×109/L, (0.33 ±0.05) ×109/L and (43.24 ±8.65) pg/ml) groups ( all P<0.05 ).The results of semi-quantitative immunohistochemical analysis showed that the p 38 protein level in control group (0.143 ±0.017) and luteolin group (0.251 ±0.021) was significantly less than that in asthmatic group ( 0.362 ±0.008 ) ( both P<0.01 ).As compared with asthmatic group , the expression of PPARγprotein markedly increased (0.247 ±0.034) in control (0.331 ±0.056) and luteolin (0.442 ±0.031) groups (all P<0.05).The level of p38 mRNA in asthmatic group (0.718 ±0.064) was significantly higher than that of control ( 0.312 ±0.052 ) and luteolin ( 0.426 ±0.067 ) groups ( all P<0.01).However, the PPARγmRNA level in asthmatic group (0.266 ±0.036) was much less than that in control (0.573 ±0.042) and luteolin (0.687 ±0.054) groups (all P<0.01).Conclusion The anti-inflammatory effects of luteolin may be associated with the regulation of PPARγexpression and p38MAPK signaling pathway in asthmatic rats.
