苦参碱对支气管哮喘小鼠气道炎症和早期气道重塑的影响
고삼감대지기관효천소서기도염증화조기기도중소적영향
Effects of matrine on airway inflammation and early airway remodeling in asthmatic mice
  • 万方数据
    中华结核和呼吸杂志 중화결핵화호흡잡지
    Chinese Journal of Tuberculosis and Respiratory Diseases
    2009年 3期 165-170 ,共6页

    雷泽林%刘晓菊%马建秀%朱骏

    뢰택림%류효국%마건수%주준

    苦参碱%哮喘%炎症%气道重塑 苦參堿%哮喘%炎癥%氣道重塑
    고삼감%효천%염증%기도중소
    Matrine%Asthma%Inflammation%Airway remodeling
    目的 观察苦参碱对支气管哮喘(简称哮喘)小鼠早期气道重塑和炎症的影响.方法 将50只BALB/C小鼠按随机数字表法分为空白对照组(A)、哮喘模型组(B)、地塞米松组(C)、苦参碱高剂量组(D,50 mg/kg)和低剂量组(E,25 mg/kg)5组.卵清白蛋白致敏建立哮喘小鼠模型,每次激发前,C、D、E组分别给予地塞米松和相应剂量的苦参碱进行灌胃;B组给予等剂量生理盐水;A组以等剂量的生理盐水致敏、激发及灌胃.肺组织切片染色,炎症细胞及黏液分泌评分、定量杯状细胞百分比,测定平滑肌面积、基底膜胶原面积.采用逆转录PCR和免疫组织化学检测转化生长因子-β1(TGF-β1)和结缔组织生长因子(CTGF)的Mrna和蛋白表达水平.采用SPSS 13.0统计软件处理,符合正态分布和方差齐性的数据采用单因素方差分析(one-way ANOVA),多组间两两比较采用Dunnet-t检验;对等级数据和不符合正态分布或方差齐性的数据则进行秩和检验(Kruskal-Wallis法),多组间两两比较采用Mann-whiteney检验;相关性采用Spearman等级相关分析.结果 A-E组炎症细胞评分(均数,四分位数)分别为1.5(1,2)、4(4,5)、2(1,3)、2(2,3)、3(2,3.3),黏液分泌评分分别为1.5(1,2)、5(4,6)、2(1,3)、2(2.5,4)、3(3,4),B组明显高于A组(X2值分别为21.3和22.6,均P<0.01),C组明显低于B组(X2值分别为13.3和15.0,均P<0.01),D、E组低于B组(X2值分别为9.1、10.9和9.8、9.7,均P<0.05);杯状细胞占上皮细胞百分比分别为(1.7±0.5)%、(54.7±15.5)%、(20.4±5.9)%、(31.7±7.6)%、(36.2±10.8)%,B组明显高于A组(t=12.0,P<0.01),C、D组明显低于B组(t值分别为7.7和5.1,均P<0.01),E组低于B组(t=4.2,P<0.05);5组平滑肌面积分别为(11.5±2.1)、(30.0±3.3)、(15.2±3.1)、(22.2±4.8)和(26.5±3.4)um2/um,B组明显高于A组(t=11.4,P<0.01),C、D、E组均明显低于B组(t值分别为9.1、4.7和2.2,均P<0.01);胶原沉积面积5组分别为(3.9±1.8)、(24.4±6.1)、(15.4±3.5)、(16.6±6.0)和(17.5±4.4)um2/um,B组明显高于A组(t=9.3,P<0.01),C、D组明显低于B组(t值分别为4.1、3.5,均P<0.01),E组低于B组(t=3.2,P<0.05);5组TGF-β1 Mrna灰度值分别为160±25、247±37、174±23、195±25、207±42,CTGF Mrna灰度值5组分别为86±8、160±24、94±10、93±14、104 4-10,B组明显高于A组(f值分别为6.1、11.6,均P<0.01),C、D、E组均明显低于B组(t值分别为3.7、2.7、5.1和10.6、8.6、10.3,均P<0.01).5组肺组织TGF-B1吸光度值分别为21±5、36±8、26±5、26±5和26±5,肺组织CTGF吸光度值分别为15±4、27±5、21±4、22±3和23±4,B组明显高于A组(t值分别为5.7和6.4,均P<0.01),c、D组明显低于B组(t值分别为3.9、3.9和3.2、2.8,均P<0.01),E组低于B组(t值分别为3.8和2.5,均P<0.05).各组小鼠胞质中TGF-β1与平滑肌面积、TGF-β1与胶原沉积面积、CTGF与平滑肌面积、CTGF与胶原沉积面积均呈正相关(r值分别为0.435、0.583、0.522和0.590,均P<0.01).结论 苦参碱能够抑制哮喘的早期气道重塑和炎症,其抑制气道重塑的可能机制与TGF-β1到CTGF的信号转导通路有关.
    목적 관찰고삼감대지기관효천(간칭효천)소서조기기도중소화염증적영향.방법 장50지BALB/C소서안수궤수자표법분위공백대조조(A)、효천모형조(B)、지새미송조(C)、고삼감고제량조(D,50 mg/kg)화저제량조(E,25 mg/kg)5조.란청백단백치민건립효천소서모형,매차격발전,C、D、E조분별급여지새미송화상응제량적고삼감진행관위;B조급여등제량생리염수;A조이등제량적생리염수치민、격발급관위.폐조직절편염색,염증세포급점액분비평분、정량배상세포백분비,측정평활기면적、기저막효원면적.채용역전록PCR화면역조직화학검측전화생장인자-β1(TGF-β1)화결체조직생장인자(CTGF)적Mrna화단백표체수평.채용SPSS 13.0통계연건처리,부합정태분포화방차제성적수거채용단인소방차분석(one-way ANOVA),다조간량량비교채용Dunnet-t검험;대등급수거화불부합정태분포혹방차제성적수거칙진행질화검험(Kruskal-Wallis법),다조간량량비교채용Mann-whiteney검험;상관성채용Spearman등급상관분석.결과 A-E조염증세포평분(균수,사분위수)분별위1.5(1,2)、4(4,5)、2(1,3)、2(2,3)、3(2,3.3),점액분비평분분별위1.5(1,2)、5(4,6)、2(1,3)、2(2.5,4)、3(3,4),B조명현고우A조(X2치분별위21.3화22.6,균P<0.01),C조명현저우B조(X2치분별위13.3화15.0,균P<0.01),D、E조저우B조(X2치분별위9.1、10.9화9.8、9.7,균P<0.05);배상세포점상피세포백분비분별위(1.7±0.5)%、(54.7±15.5)%、(20.4±5.9)%、(31.7±7.6)%、(36.2±10.8)%,B조명현고우A조(t=12.0,P<0.01),C、D조명현저우B조(t치분별위7.7화5.1,균P<0.01),E조저우B조(t=4.2,P<0.05);5조평활기면적분별위(11.5±2.1)、(30.0±3.3)、(15.2±3.1)、(22.2±4.8)화(26.5±3.4)um2/um,B조명현고우A조(t=11.4,P<0.01),C、D、E조균명현저우B조(t치분별위9.1、4.7화2.2,균P<0.01);효원침적면적5조분별위(3.9±1.8)、(24.4±6.1)、(15.4±3.5)、(16.6±6.0)화(17.5±4.4)um2/um,B조명현고우A조(t=9.3,P<0.01),C、D조명현저우B조(t치분별위4.1、3.5,균P<0.01),E조저우B조(t=3.2,P<0.05);5조TGF-β1 Mrna회도치분별위160±25、247±37、174±23、195±25、207±42,CTGF Mrna회도치5조분별위86±8、160±24、94±10、93±14、104 4-10,B조명현고우A조(f치분별위6.1、11.6,균P<0.01),C、D、E조균명현저우B조(t치분별위3.7、2.7、5.1화10.6、8.6、10.3,균P<0.01).5조폐조직TGF-B1흡광도치분별위21±5、36±8、26±5、26±5화26±5,폐조직CTGF흡광도치분별위15±4、27±5、21±4、22±3화23±4,B조명현고우A조(t치분별위5.7화6.4,균P<0.01),c、D조명현저우B조(t치분별위3.9、3.9화3.2、2.8,균P<0.01),E조저우B조(t치분별위3.8화2.5,균P<0.05).각조소서포질중TGF-β1여평활기면적、TGF-β1여효원침적면적、CTGF여평활기면적、CTGF여효원침적면적균정정상관(r치분별위0.435、0.583、0.522화0.590,균P<0.01).결론 고삼감능구억제효천적조기기도중소화염증,기억제기도중소적가능궤제여TGF-β1도CTGF적신호전도통로유관.
    Objective To observe the influence of matrine on airway inflammation and early airway remodeling in asthmatic mice. Methods Fifty BALB/c mice were randomly divided into 5 groups: a normal control group (A), an asthmatic group (B), a dexamethasone (DXM) group (C, 2 mg/kg), a high-dose matrine group (D, 50 mg/kg) and a low-dose matrine group (E, 25 mg/kg). The mice model of asthma in the B, C, D, and E groups was established by ovalbumin (OVA) intraperitoneal injections and aerosolizations. Intra-gastric administrations of different medications in C, D, E groups and 0. 9% sodium chloride in B group were carried out 1 hour before provocation. 0.9% sodium chloride was used for intraperitoneal injection, aerosolization and intra-gastric administration in group A. The lung tissue slices were stained, and then the grade of inflammation around the wall of bronchi, mucous secretion, and the percentage of goblet-cells were counted. The areas of bronchial smooth muscle and of collagen deposition in airway wall were analyzed. The transcriptions and protein expressions of transforming growth factor-?,1> (TGF-?,1>) and connective tissue growth factor (CTGF) were measured respectively by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry. Results In the A, B, C, D, E groups, the grades of inflammation were 1.5 (1,2), 4 (4,5), 2 (1,3), 2 (2,3), 3 (2,3. 3), respectively; the degrees of mucous secretions were 1.5 (1,2), 5 (4,6), 2 (1,3), 2(2.5,4), 3 (3,4), respectively. These airway inflammatory parameters in group B were significantly higher than in group A (X2=21.3, 22.6, P all<0.01), while they were remarkably decreased in group C compared to group B (X2=13.3, 15.0, Pall <0.01). These parameters in group D and group E were also lower than those in group B (X2=9.1, 10.9; 9.8, 9.7; P all<0.05). The percentage of goblet cells in airway epithelium was (1.7?.5)%, (54.7?5.5)%, (20.4?.9)%, (31.7?.6)% and (36.2?0.8)%, respectively; it was significantly higher in group B than in group A (t=12.0, P<0.01), and remarkably lower in groups C and D than in group B (t=7.7, 5.1, P all<0.01), and lower in group E than in B group (t=4.2, P < 0.05). In these 5 groups, the area of bronchial smooth muscle was (11.5?.1) um2/um, (30.0?.3) um2/um, (15.2?.1) um2/um, (22.2?.8) um2/um and (26.5?.4) um2/um, respectively; it was significantly higher in group B than in group A (t=11.4, P<0.01), and remarkably lower in groups C, D and E than in group B (t=9.1, 4.7, 2.2, P all<0.01). The area of collagen deposition was (3.9?.8) um2/um, (24.4?.1) um2/um, (15.4?.5) um2/um, (16.6?.0) um2/um and (17.5?.4) um2/um, respectively; it was also significantly higher in group B than in group A (t=9.3, P<0.01), and remarkably lower in groups C and D than in group B (t=4.1, 3.5, P all <0.01), and lower in group E than in B group (t=3.2, P<0.05). The mRNA levels of TGF-?,1> were 160?5,247?7, 174?3, 195?5 and 207?2, respectively, and those of CTGF were 86?, 160?4, 94?0, 93?4 and 104?0, respectively in the 5 groups. The levels were remarkably increased in group B, as compared to group A (t=6.1, 11.6, P all<0.01), and the levels in groups C, D and E were remarkably decreased, as compared to group B, the difference being significant (t=3.7, 2.7, 5.1; 10.6, 8.6, 10.3; P all <0.01). The protein level of TGF-?,1> in lung tissues was 21?, 36?, 26?, 26? And 26?, respectively, and that of CTGF was 15?, 27?, 21?, 22? And 23?, respectively in the 5 groups. The levels in B group were significantly increased, as compared to group A (t=5.7, 6.4, P all <0.01), and those in groups C and D were significantly decreased (t=3.9, 3.9; 3.2, 2.8, P all <0.01), and that in group E was also lower (t=3.8, 2.5, P all<0.05), as compared to group B. In all the groups, the protein levels of TGF-?,1> and CTGF were positively correlated with the area of bronchial smooth muscle and with the area of collagen deposition (r=0.435, 0.583, 0.522, 0.590, P all <0.01). Conclusions Matrine inhibited airway inflammation and early airway remodeling in asthmatin mice.the signal transduction of TGF-β1 and CTGF maybe involved.
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