复方薤白胶囊治疗肺动脉高压抗内皮细胞凋亡的研究
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摘要
目的:慢性阻塞性肺病(COPD)肺动脉高压(PAH)的发病机制为,肺血管内皮功能失调、低氧性肺血管收缩、肺血管重构,而内皮细胞损伤是PAH的起始环节。复方薤白胶囊治疗COPD并PAH疗效显著,并于2004年取得国家发明专利权,专利号ZL02160139.9。本课题研究复方薤白提取液对过氧化氢(H_2O_2)诱导的凋亡内皮细胞的保护作用,及对凋亡相关基因及信号转导通路的影响,以探讨其抗凋亡机制。
方法:体外培养人脐静脉内皮细胞(HUVEC),以过氧化氢(H_2O_2)行内皮细胞凋亡造模,以MTT法筛选H_2O_2造模浓度及实验时间。设立5个实验组:正常组(HUVEC)、对照组(HUVEC+H_2O_2)、中药低、中、高剂量组(HUVEC+H_2O_2+复方薤白胶囊提取液3μg/ml、30μg/ml、300μg/ml)。分别以MTT法检测各组细胞增殖情况、光镜下形态学及Hoechst染色观察细胞凋亡、流式细胞检测细胞凋亡率、以Western Blot检测抗凋亡基因Bcl-2蛋白表达变化、罗丹明123(Rh123)染色检测线粒体膜电位、免疫组织化学法检测信号转导通路Caspase-3、NF—KB的表达。
结果:
1.造模结果:200μmol/l的H_2O_2为最佳造模浓度,该浓度作用HUVEC 2小时造模成功。造模后24小时作为最佳实验研究时间。
2.MTT法检测细胞增殖结果显示:对照组HUVEC吸光度(OD值)明显降低,细胞增殖率仅9.29%(P＜0.01)。经不同浓度复方薤白胶囊提取液处理后,HUVEC的OD值、增殖率逐渐提高,分别为14.77%、31.98%、41.45%,以中、高剂量组增殖最为明显,与对照组比P＜0.01,具有明显的统计学意义。
3.Hoechst染色观察细胞凋亡结果示:正常组凋亡率为2.0%,对照组见大量凋亡细胞,凋亡率为32.17%(P＜0.01)。中药低、中、高浓度组的凋亡率分别为28.33%、20.17%、13.33%。其中以中、高剂量组与对照组比P＜0.01,具有明显的统计学意义。
4.流式细胞检测细胞凋亡率结果示:正常组细胞凋亡率为0.37%。对照组达14.39%(P＜0.01)。低、中、高中药组HUVEC细胞凋亡率分别为9.10%、7.91%、6.48%。与对照组比P＜0.01,具有明显的统计学意义。
5.Western Blot检测Bcl-2结果示:正常组为0.673,对照组为0.438,低、中、高中药组分别为0.572、0.613、0.628,各中药组的Bcl-2基因表达量较对照组明显增高。
6.罗丹明123(Rh123)染色检测线粒体膜电位结果显示:正常组细胞发出明显绿色荧光,而对照组荧光强度微弱。而经复方薤白胶囊提取液处理后,各中药组随药物浓度的增加,绿色荧光逐渐增多,其中以中、高剂量组为明显。
7.Caspase-3免疫组织化学结果示:对照组的胞浆中见明显紫红色颗粒,各中药组胞浆中紫红色颗粒逐渐减少,且与药物浓度呈正相关。正常组阳性细胞为1.83%,对照组高达20.67%(P＜0.01),低、中、高中药组的阳性细胞数分别为19.33%、14.33%、5.33%,其中以中、高剂量组与对照组相比,P＜0.01,具有明显的统计学意义。
8.NF—KB免疫组织化学结果示:对照组的胞浆中见明显紫红色颗粒,各中药组胞浆中紫红色颗粒逐渐减少,且与药物浓度呈正相关。正常组阳性细胞为3.33%,对照组达18.5%,P＜0.01,低、中、高中药组分别为16.67%、12.33%、6.67%,其中以中、高剂量组与对照组相比,P＜0.01,具有明显的统计学意义。
结论:
复方薤白胶囊对氧化损伤所致凋亡的内皮细胞具有保护作用,能抗内皮细胞凋亡。其抗凋亡的机理为提高抗凋亡基因Bcl-2水平的表达。而该基因表达的上调,与该药改变了内皮细胞的线粒体膜的通透性,增强线粒体跨膜电位的表达,下调了Caspase-3及NF-κB的信号转导通路有关。由此我们认为,复方薤白胶囊治疗COPD、PAH的作用机制是,通过多个作用靶点保护内皮细胞,起到抗凋亡作用。
Objective:The pathogenesis of chronic obstructive pulmonary disease(COPD) and pulmonary artery hypertension(PAH) are pulmonary endothelial dysfunction,hypoxic pulmonary vasoconstriction,vascular remodelling.Recent investigation proves that the damage of endothelial cell is the initiation of PAH.Compound Macrostem Onion Capsule (CMOC),which got patent right in 2004,NO.ZL02160139.9,is an effective formula for patients with COPD and PAH.This project study was designed to assess the protective effect of CMOC on hydrogen dioxided(H_2O_2) induced apoptosis in human umbilical vascular endothelial cell(HUVEC),apoptosis-related genes expression and signal transduction factors, to reveal the mechanism of anti-apoptosis of CMOC on Endothelial Cell.
Methods:To establish the apoptotic models of HUVEC in vitro,using MTT assay to decide the best dose and the best time of H_2O_2.HUVEC were divided into five experimental groups:normal group,control group,CMOC group with three concentrations of 3μg/ml, 30μg/ml and 300μg/ml respectively MTT assay was used to measure the proliferation of the HUVEC.The apoptotic characteristics were detected by means of flow cytometry and Hoechst stain.Genes bcl-2 were detected by Western blotting.Using Rh123 dying to detect the expression of mitochondrial membrane potential.Using immunohistochemistry staining to detect the expression of caspase-3 and NF—KB.
Results:
1.The model of apoptotic were established by H_2O_2.The result showed that the best concentration of H_2O_2 was 200μmol/l,and the best duration to establish apoptotic model was 24 hours.
2.MTT assay result:Cell optical density were lower than normal group and the proliferation rate of the cells was 9.29%in the control group(P＜0.01).When treated by CMOC,the OD value were increased,and the proliferation rate of the cells were14.77%, 31.98%and 41.45%in CMOC groups respectively which increased with the increase of concentration.There were apparent statistical significance in the middle and high concentration groups,as compared with the control group,P＜0.01.
3.Hoechst dying result:Apoptosis cell were seldom found in the normal group the apoptosis rate was 2.0%.In the control group a mass of apoptosis were detected and the apoptosis rate was 32.17%.After treated with CMOC,apoptosis cells were reduced in number.The apoptosis rate were 28.33%,20.17%,13.33%in CMOC groups respectively,in direct proportion to concentration.There were apparent statistical significance in the middle and high concentration groups,as compared with the control group,P＜0.01.
4.Flow cytometry assay result:The cellular apoptosis rate were 0.37%in the normal group,14.38%in the control group,9.1%,7.9%and 6.48%in CMOC groups respectively which were proportional to the increase of concentration.There were apparent statistical significance compared with the control group,P＜0.01.
5.Western blot result:The value of Bcl-2 expression were 0.673 in normal group, 0.438 in control group,0.572,0.613 and 0.628 in CMOC groups respectively,in direct proportion to concentration.It shows that the anti-apoptosis gene expression was increased after CMOC treatment.
6.Mitochondrial membrane potential result:In the normal group Rh123 dying emitted apparent green florescent light,while in the control group the florescent light were feeble. After treated with CMOC,the florescent light increased in intensity and were positively correlated with concentration,especially in the middle and high concentration groups.
7.Caspase-3 immunohistochemistry result:The prunosus particle in the control group were the most apparent,after treated with CMOC,the prunosus particle reduced in number. The positive cells were 1.83%in normal group,20.67%in control group,19.33%,14.33% and 5.33%in CMOC groups respectively,in direct proportion to concentration.The most apparent statistical significance were found in the middle and high concentration groups,as compared with the control group,P＜0.01.
8.NF—KB immunohistochemistry result:The prunosus particle in the control group were the most apparent,but after treated with CMOC,the prunosus particle reduced in number.The positive cells were 3.33%in normal group,18.5%in control group, 16.67%,12.33%and 6.67%in CMOC groups respectively,which were again in direct proportion to concentration.The most apparent statistical significance were found in the middle and high concentration groups,as compared with the control group,P＜0.01.
Conclusions:
These results indicate that Compound Macrostem Onion Capsule(CMOC) has the protective effect on the H_2O_2 induced cellular apoptosis in HUVEC,and has the function of anti-apoptosis.Its mechanism lies in its effect in increasing the expression of Bcl-2.The expression of this gene may change the permeability of mitochondrial,enhance the expression of membrane potential,reduced Caspae-3 and NF-KB signal pathway,and suppress caspase-3 induced cell apoptosis.Thus we think CMOC can protect HUVEC via many targets and can prevent apoptosis.
方法:体外培养人脐静脉内皮细胞(HUVEC),以过氧化氢(H_2O_2)行内皮细胞凋亡造模,以MTT法筛选H_2O_2造模浓度及实验时间。设立5个实验组:正常组(HUVEC)、对照组(HUVEC+H_2O_2)、中药低、中、高剂量组(HUVEC+H_2O_2+复方薤白胶囊提取液3μg/ml、30μg/ml、300μg/ml)。分别以MTT法检测各组细胞增殖情况、光镜下形态学及Hoechst染色观察细胞凋亡、流式细胞检测细胞凋亡率、以Western Blot检测抗凋亡基因Bcl-2蛋白表达变化、罗丹明123(Rh123)染色检测线粒体膜电位、免疫组织化学法检测信号转导通路Caspase-3、NF—KB的表达。
结果:
1.造模结果:200μmol/l的H_2O_2为最佳造模浓度,该浓度作用HUVEC 2小时造模成功。造模后24小时作为最佳实验研究时间。
2.MTT法检测细胞增殖结果显示:对照组HUVEC吸光度(OD值)明显降低,细胞增殖率仅9.29%(P＜0.01)。经不同浓度复方薤白胶囊提取液处理后,HUVEC的OD值、增殖率逐渐提高,分别为14.77%、31.98%、41.45%,以中、高剂量组增殖最为明显,与对照组比P＜0.01,具有明显的统计学意义。
3.Hoechst染色观察细胞凋亡结果示:正常组凋亡率为2.0%,对照组见大量凋亡细胞,凋亡率为32.17%(P＜0.01)。中药低、中、高浓度组的凋亡率分别为28.33%、20.17%、13.33%。其中以中、高剂量组与对照组比P＜0.01,具有明显的统计学意义。
4.流式细胞检测细胞凋亡率结果示:正常组细胞凋亡率为0.37%。对照组达14.39%(P＜0.01)。低、中、高中药组HUVEC细胞凋亡率分别为9.10%、7.91%、6.48%。与对照组比P＜0.01,具有明显的统计学意义。
5.Western Blot检测Bcl-2结果示:正常组为0.673,对照组为0.438,低、中、高中药组分别为0.572、0.613、0.628,各中药组的Bcl-2基因表达量较对照组明显增高。
6.罗丹明123(Rh123)染色检测线粒体膜电位结果显示:正常组细胞发出明显绿色荧光,而对照组荧光强度微弱。而经复方薤白胶囊提取液处理后,各中药组随药物浓度的增加,绿色荧光逐渐增多,其中以中、高剂量组为明显。
7.Caspase-3免疫组织化学结果示:对照组的胞浆中见明显紫红色颗粒,各中药组胞浆中紫红色颗粒逐渐减少,且与药物浓度呈正相关。正常组阳性细胞为1.83%,对照组高达20.67%(P＜0.01),低、中、高中药组的阳性细胞数分别为19.33%、14.33%、5.33%,其中以中、高剂量组与对照组相比,P＜0.01,具有明显的统计学意义。
8.NF—KB免疫组织化学结果示:对照组的胞浆中见明显紫红色颗粒,各中药组胞浆中紫红色颗粒逐渐减少,且与药物浓度呈正相关。正常组阳性细胞为3.33%,对照组达18.5%,P＜0.01,低、中、高中药组分别为16.67%、12.33%、6.67%,其中以中、高剂量组与对照组相比,P＜0.01,具有明显的统计学意义。
结论:
复方薤白胶囊对氧化损伤所致凋亡的内皮细胞具有保护作用,能抗内皮细胞凋亡。其抗凋亡的机理为提高抗凋亡基因Bcl-2水平的表达。而该基因表达的上调,与该药改变了内皮细胞的线粒体膜的通透性,增强线粒体跨膜电位的表达,下调了Caspase-3及NF-κB的信号转导通路有关。由此我们认为,复方薤白胶囊治疗COPD、PAH的作用机制是,通过多个作用靶点保护内皮细胞,起到抗凋亡作用。
Objective:The pathogenesis of chronic obstructive pulmonary disease(COPD) and pulmonary artery hypertension(PAH) are pulmonary endothelial dysfunction,hypoxic pulmonary vasoconstriction,vascular remodelling.Recent investigation proves that the damage of endothelial cell is the initiation of PAH.Compound Macrostem Onion Capsule (CMOC),which got patent right in 2004,NO.ZL02160139.9,is an effective formula for patients with COPD and PAH.This project study was designed to assess the protective effect of CMOC on hydrogen dioxided(H_2O_2) induced apoptosis in human umbilical vascular endothelial cell(HUVEC),apoptosis-related genes expression and signal transduction factors, to reveal the mechanism of anti-apoptosis of CMOC on Endothelial Cell.
Methods:To establish the apoptotic models of HUVEC in vitro,using MTT assay to decide the best dose and the best time of H_2O_2.HUVEC were divided into five experimental groups:normal group,control group,CMOC group with three concentrations of 3μg/ml, 30μg/ml and 300μg/ml respectively MTT assay was used to measure the proliferation of the HUVEC.The apoptotic characteristics were detected by means of flow cytometry and Hoechst stain.Genes bcl-2 were detected by Western blotting.Using Rh123 dying to detect the expression of mitochondrial membrane potential.Using immunohistochemistry staining to detect the expression of caspase-3 and NF—KB.
Results:
1.The model of apoptotic were established by H_2O_2.The result showed that the best concentration of H_2O_2 was 200μmol/l,and the best duration to establish apoptotic model was 24 hours.
2.MTT assay result:Cell optical density were lower than normal group and the proliferation rate of the cells was 9.29%in the control group(P＜0.01).When treated by CMOC,the OD value were increased,and the proliferation rate of the cells were14.77%, 31.98%and 41.45%in CMOC groups respectively which increased with the increase of concentration.There were apparent statistical significance in the middle and high concentration groups,as compared with the control group,P＜0.01.
3.Hoechst dying result:Apoptosis cell were seldom found in the normal group the apoptosis rate was 2.0%.In the control group a mass of apoptosis were detected and the apoptosis rate was 32.17%.After treated with CMOC,apoptosis cells were reduced in number.The apoptosis rate were 28.33%,20.17%,13.33%in CMOC groups respectively,in direct proportion to concentration.There were apparent statistical significance in the middle and high concentration groups,as compared with the control group,P＜0.01.
4.Flow cytometry assay result:The cellular apoptosis rate were 0.37%in the normal group,14.38%in the control group,9.1%,7.9%and 6.48%in CMOC groups respectively which were proportional to the increase of concentration.There were apparent statistical significance compared with the control group,P＜0.01.
5.Western blot result:The value of Bcl-2 expression were 0.673 in normal group, 0.438 in control group,0.572,0.613 and 0.628 in CMOC groups respectively,in direct proportion to concentration.It shows that the anti-apoptosis gene expression was increased after CMOC treatment.
6.Mitochondrial membrane potential result:In the normal group Rh123 dying emitted apparent green florescent light,while in the control group the florescent light were feeble. After treated with CMOC,the florescent light increased in intensity and were positively correlated with concentration,especially in the middle and high concentration groups.
7.Caspase-3 immunohistochemistry result:The prunosus particle in the control group were the most apparent,after treated with CMOC,the prunosus particle reduced in number. The positive cells were 1.83%in normal group,20.67%in control group,19.33%,14.33% and 5.33%in CMOC groups respectively,in direct proportion to concentration.The most apparent statistical significance were found in the middle and high concentration groups,as compared with the control group,P＜0.01.
8.NF—KB immunohistochemistry result:The prunosus particle in the control group were the most apparent,but after treated with CMOC,the prunosus particle reduced in number.The positive cells were 3.33%in normal group,18.5%in control group, 16.67%,12.33%and 6.67%in CMOC groups respectively,which were again in direct proportion to concentration.The most apparent statistical significance were found in the middle and high concentration groups,as compared with the control group,P＜0.01.
Conclusions:
These results indicate that Compound Macrostem Onion Capsule(CMOC) has the protective effect on the H_2O_2 induced cellular apoptosis in HUVEC,and has the function of anti-apoptosis.Its mechanism lies in its effect in increasing the expression of Bcl-2.The expression of this gene may change the permeability of mitochondrial,enhance the expression of membrane potential,reduced Caspae-3 and NF-KB signal pathway,and suppress caspase-3 induced cell apoptosis.Thus we think CMOC can protect HUVEC via many targets and can prevent apoptosis.
引文
1.Fabbri LM,Hurd SS.Global strategy for the diagnosis,management and prevention of COPD:2003 update[J].Eur Respir J,2003,22(1):1-2.
2.Hoeper MM,Mayer E,Simonneau Get al.Chronic thromboembolic pulmonary hypertension [J].Circulation,2006,113:2 011.
3.COOL CD,STEWART JS,WERAHERA P,et al.Three dimensional reconstruction of pulmonary arteries in plexiform pulmonary hypertension using cell specific markers:evidence for adynamic and heterogeneous process of pulmonary endothelial cell growth[J].A m J Pat hol,1999,155:411-419.
4.Wright JL,Levy RD,Churg A.Pulmonary hypertension in chronic obstructive pulmonary disease:current theories of pathogenesis and their implications for treatment[J].Thorax,2005,60(7):605-609.
5.Arao T,Takabatake N,Sata M,et al.In vivo evidence of endothelial injury in chronic obstructive pulmonary disease by lung scintigraphic assessment of(123) Imetaiodobenzylguanidine.[J].J Nucl Med,2003,44(11):1747-1754.
6.Bagatcheva NV,Garcia JG,Verin AD.Molecular mechanism ofthrombin-induced endothelial cell permeability[J].Biochemistry,2002,67:75-84.
7.陈瑞芬,周光德,曹文军,等.野百合碱诱导实验性肺动脉高压病理形态观察[J].电子显微学报,2002,21(1):1-4.
8.席思川,车东媛,张婉蓉,等.缺氧性肺动脉高压与肺腺泡内动脉构型重组[J].中国循环杂志,1995,(5):287-290.
9.王良兴,陈少贤,徐正介,等.低氧高二氧化碳环境下肺动脉内皮结构变化与肺动脉高压的关系[J].心肺血管病杂志,2000,19(3):220-223.
10.陈小客,邹霞英,辛达临.缺氧性肺动脉高压大鼠肺血管内皮细胞的变化[J].中国病理生理杂志,1996,12(6):608-611.
11.Voelkel NF,Cool CD.Pulmonary vascular involvement in chronic obstructive pulmonary disease[J].Eur Respir J Suppl,2003,46(suppl):28s-32s.
12.Furchgott RF,Zawadzki JV.The obligatory role of endothelial cells in the relation of arterial smooth muscle by acetylcheoling[J].Nature,1980,288(5789):373-376.
13.Voelkel NF,Cool C.Pathology of pulmonary hypertension[J].Cardiol Clin,2004,22:343-351.
14.Taraseviciene-Stewart L,Kasahare Y,Alger L,et al.Inhibition of the VEGF receptor 2 combined with chronic hypoxia causes cell death-dependent pulmonary endothelial cell proliferation and severe pulmonary hypertension[J].FASEB J,2001,15;427-438.
15.Hoshikawa Y,Ono S,Suzuki S,et al.Generation of oxidative stresss contributes to the development of pulmonary hypertension induced by hypoxia[J].J Appl Physionl,2001,90(4):1299-1306.
16.杨洁,易静,汤雪明.氧化修饰在调控细胞凋亡信号转导中的作用[J].细胞生物学杂志, 2005,27:121-126.
17.毛伟平,许峰.细胞凋亡过程中的信号转导[J].环境与职业医学,2004,21(1):62-65.
18.朱晓东.凋亡通路及其调控研究进展[J].国外医学,生理、病理科学与临床分册,2002,22(3):248-251.
19.Adams JM,Cory S.Life or death decisions by the Bcl-2 protein family[J].Trends Biochem Sci,2001,26(1):61-66.
20.KUWANA T,MACKEY M R,PERKINS G,et al.Bid,bax,and lipids cooperate to form supra molecular openings in the outer mitochondrial membrane[J].Cell,2002,111(3):331-342.
21.PETIT P X,ZAMZAMI N,VAYSSIERE J L,et al.Implication ofmitochondria in apoptosis[J].Mol Cell Biochem,1997,174(1-2):185-188.
22.NEWMEYER D D,FERGUSON-MILLER S.Mitochondria:Releasing power for life and unleashing machineries of death[J].Ce11,2003,112(4):481-490.
23.RAO RV,HERMEL E,CASTRO-OBREGON S,et al.Coupling endoplasmic reticulum stress to the cell death program:mechanism ofcaspase activation[J].J Biol Chem,2001,276(36):33869-33874.
24.Wallach D,Vorfolmeev EE,Malinin NL,et al.Tumor necrosis factor receptor and fas signaling mechanisms[J].Annu Rev Immunol,1999,17:331-367.
25.Karin M,Lin A.NF-kappa B at the crossroads of life and death.Nat Immunol.2002,3(3):221-227.
26.Michiels C,Minet E,Mottet D,et al.Regulation of gene expression by oxygen:NF-kappaB and HIF-1,two extremes[J].Free Radio Biol Med,2002,33(9):1231-1242.
27.Wang CY,Guttridge DC,Mayo MW,et al.NF-kappa B induces expression of the Bcl-2 homologue Al/Bfl-1 to preferentially suppress chemotherapy-induced apoptosis[J].Mol Cell Biol,1999,19:5923-5929.
28.Wang CY,Mayo MW,Komeluk RG,et al.NF-kappa B antiapoptosis:induction of TRAF1 and TRAF2 and c-IAP1 and c-IAP2 to suppress Caspase-8 activation[J].Science,1998,281:1680-1683.
29.Susin SA,Zamzami N,Castedo M,et al.The central executioner of apoptosis:multiple connections between protease activation and mitochondrial in Fas/APO-1/CD95-and ceramide-induced apoptosis [J].J Exp Med,1997,186:25-37.
30.王海燕,王来栓.细胞凋亡通路研究进展[J].国外医学生理病理科学与临床分册,2003,23(5):490-492.
31.Nakamura K,Bossy Wetzel E,Bums K,et al.Changes in endoplasmic reticulum luminal environment affect cell sensitivity to apoptosis[J].J Cell Biol,2000,150(4):731-740.
32.Rao RV,Hermel E,Castro-Obregon S,et al.Coupling endoplasmic reticulum stress to the cell death program.Mechanism of caspase activation[J].J Biol Chem,2001,276(36):33869-33874.
33.Pinton P,Ferrari D,Papizzi Z,et al.The Ca~(2+) concentration of the endoplasmic reticulum is a key determinant of ceramide-induced apoptosis:significance for the molecular mechanism of Bcl-2 action [J]. EMBo J,2001,20:2690-2701.
34. McConkey DJ , Nutt LK. Calciumflux measurements in apoptosis [J ]. Methods Cell Biol, 2001,66(2):229-246.
35. Nutt LK, Pataer A, Pahler J, et al. Bax and Bak promote apoptosis by modulating endoplasmic reticular and mitochondrial Ca~(2+) stores [J ]. J Biol Chem , 2002,277(11):9219-9225.
36. Breckenridge DG, Stojanovic M, Marcellus RC , et al. Caspase cleavage product of BAP31 induces mitochondrial fission through endoplasmic reticulum calcium signals , enhancing cytochrome release to the cytosol [J ]. J Cell Biol, 2003,160(7):l 115-1127.
37. Fujimoto K, Matsuzawa Y, Yamaguchi S. Benefits of oxygen on exercise performance and pulmonary hemodynamics in patients with COPD with mild hypoxemia[J ]. Chest ,2002 ,122 :457—463.
38. Franz IW, Van Der Meyden J, Schaupp S , et al. The effect of amlodipine on exercise—induced pulmonary hypertension and right heart function in patients with chronic obstructive pulmonary disease [J]. Z Kardiol ,2002,91 :833—839.
39. Borst MM ,Leschke M , Koing U ,et al. Repetitive hemodilution in chronic obstructive pulmonary disease and pulmonary hypertension :effects on pulmonary hemodynamics , gas exchange,and exercise capacity [J]. Respiration ,1999 ,66 :225—232.
40. Weg IL , Rossoff L , McKeon K. Development of pulmonary hypertension after lung volume reduction surgery [J]. Am J Respir Crit Care Med ,1999 ,159 :552-556.
41. Oswald—Mammosser M, Kessler R, Massard G, et al. Effect of lung volume reduction surgery on gas exchange and pulmonary hemodynamics at rest and during exercise [J]. Am J Respir Crit Care Med,1998 ,158:1020-1025.
42. Zhao YD, Courtman DW, Deng Y, et al. Rescue of monocrotaline induced pulmonary arterial hypertension using bone marrow-derived endothelial-like progenitor cells: efficacy of combined cell and eNOS gene therapy in established disease [J]. Circ Res, 2005.96:442-450.
43. Nagaya N, Kangawa K, Kanda M, et al. Hybrid cell-gene therapy for pulmonary hypertension based on phagocyteosing action of endpthelial progenitor cells [J]. Circulation, 2003,108:889-895.
44. KENTARO H, JUN F , YOSHIKO M, et al. Bone marrow derived cells cont ribute to pulmonary vascular remodeling in hypoxia-Induced pulmonary hypertension [J ]. Chest, 2005,127:1793-1798.
45. KIMIO S , YU TA KA K, MA KOTO N , et al. Important role of endogenous erythropoietin system in recruitment of endothelial progenitor cells in hypoxia-induced pulmonary hypertension in mice [J].Circulation, 2006 ,113 :1442-1450.
46. NAGA YA N , KANGAWA K, KANDA M , et al. Hybrid cell-gene therapy for pulmonary hypertension based on phagocytosing action of endothelial progenitor cells [J].Ciculation,2003,108 :889-895.
47.周仲瑛.周仲瑛临床经验辑要[M].北京:中国医药科技出版社,1998.1(1):36.
48.李素云,吴其标.曹世宏教授论治慢性阻塞性肺疾病经验选粹[J].中医药学刊.2002,20(1):28.
49.奚肇庆,刘学华,冷报浪,等.通阳宣痹法治疗慢性阻塞性肺病急性发作期临床研究[J].中国中医急症,2005,14(11):1025-1027.
50.奚肇庆,蒋荫,居文政,等.复方薤白胶囊治疗慢性阻塞性肺病36例临床与实验研究[J].中医杂志,2000,4(41):218-220.
51.奚肇庆.通阳宣痹法治疗支气管哮喘急性发作期的临床研究[J].江苏中医,1999,20(2):14-15.
52.冷报浪,奚肇庆,曹世宏,等.复方薤白胶囊对慢性支气管炎大鼠血小板活化因子及一氧化氮的影响[J].现代中西医结合杂志,2006,15(10):1293-1299.
53.孙菊光,奚肇庆,姜静,等.复方薤白胶囊对野百合碱诱导的肺动脉高压大鼠病理的影响[J].南京中医药大学学报(自然科学版),2006,22(3):173-176.
54.孙菊光,奚肇庆,姜静,等.复方薤白胶囊对野百合碱诱导肺动脉高压大鼠SOD活性、MDA、HYP含量的影响[J].实用中医药杂志,2005,21(12):711-713.
55.孙菊光,奚肇庆,姜静,等.复方薤白胶囊对野百合碱诱导肺动脉高压大鼠ET、TNF α含量的影响[J].东南大学学报(医学版),2006,25(5):337-340.
56.姜静,尚宁,奚肇庆,等.复方薤白胶囊对野百合碱诱导肺动脉高压大鼠血浆中花生四烯酸代谢产物的影响[J].中华结核和呼吸杂志,2005,28(8):534-536.
57.奚肇庆,姜静,孙菊光,等.复方薤白胶囊对野百合碱诱导大鼠肺动脉高压的抑制作用[J].中医杂志,2006,47(10):781-783.
58.王剑蓉,朱云华.复方薤白胶囊治疗实验性慢性支气管炎的病理观察[J].南京中医药大学学报(自然科学版),2002,18(4):224-225.
59.陈丽萍,白旭东,于国梁.高分辨率超声观察薤自制剂对高脂血症的治疗作用[J].中国老年学杂志,2002,22(5):182.
60.谭可安.薤白的临床疗效观察[J].白求恩医科大学学报,1989.15(2):211.
61.陈滴.薤白对家兔体内前列腺素El的影响[J].白求恩医科大学学报,1989.15(1):91.
62.孟庆国,朱庆磊,邓淑娥,等.薤白水提物对羟自由基的清除作用[J].潍坊医学院学报,1998,20(1):66.
63.沈树人.“薤白桂皮汤”治疗毛细支气管炎合并肠炎50例[J].中西医结合实用临床急症,1997,10(4):462-463.
64.李向红.薤白的提取物抗氧化作用研究[J].中药材,1994,17(11):34.
65.吴洪元.薤白的炮制研究[J].中药材,1995,18(4):192.
66.吴波,陈思维,王伟敏,等.瓜蒌薤白白酒汤提取物抗心肌缺血缺氧及最佳处方的筛选[J].中草药,2000,31(11):844.
67.张卿,高尔,侯琦,等.薤白挥发油对S180荷瘤小鼠免疫功能的影响[J].潍坊医学院学报,2002,24(2):94.
68.阴健,郭力弓主编.中药现代研究与临床应用[M].北京:学院出版社,1993,260-263.
69.吴波,王敏伟,陈思维,等.瓜蒌提取物对离体家兔胸主动脉条收缩的影响[J].沈阳药科大学学报,1999,16(1):24-27.
70.贝伟剑.瓜蒌薤白汤的药理作用[J].中国医药学报,1989,4(5):341.
71.邵春丽,王世久,王进,等.瓜蒌皮抗缺氧作用的研究[J].沈阳药科大学学报,1998,15(1):38-40.
72.向邦平,金善炜,傅桂香,等.瓜蒌蛋白Ⅱ.瓜萎蛋白部分化学结构的初步测定[J].化学学报,1998.56:302-303.
73.李仪奎.中药药理学[M].北京:中国中医药出版社,1992:157.
74.沈雅琴,张明发.半夏的镇痛、抗溃疡和抗血栓作用的形成[J].中国生化药物杂志,1998,19(3):141.
75.刘继林.水半夏与半夏部分药理作用的对比研究[J].成都中医药学院学报,1989,12(2):41.
76.洪行球.半夏降血脂作用研究[J].浙江中医学院学报,1995,19(2):28-29.
77.王光明,周蓉.半夏的中药药理研究进展[J].中医药导报,2007,13(2):97-99.
78.Marin Nero JA,Maciel BC,Secches AL,et al.Cardiovascular effects of berberine in patients with severe congestive heart failure[J].Clin Car diol,1988,11(4):253-260.
79.周祖玉,孙爱民,徐建国,等.黄连素对离体灌流心脏的能量保存作用[J].华西医科大学学报,2002,33(3):431-433.
80.Zeng XH,Zeng XJ,Li YY.Efficacy and safety of berberine for congestive heart failure secondary to ischemic or idiopathic dilated cardiomyopathy[J].AmJ Cardiol,2003,92(2):173-176.
81.袁拯忠,朱陵群,庞鹤等.黄连解毒汤有效成分对缺氧/复氧时脑微血管内皮细胞的保护作用[J].中国中药杂志,2007,32(3):249-252.
82.崔学军.黄连及其有效成分的药理研究进展[J].中国药师,2006,9(5):469-470.
83.王海燕,李玉清,王启会.黄连素抗肿瘤作用研究进展[J].中成药,2007,29(10):1500-1502.
84.陈彬,张琪伟.瓜蒌薤白药对大鼠心功能及血流变的影响[J].南京中医药大学学报,1996,12(2):26.
85.徐长胜.瓜蒌薤白方加味止咳祛痰作用的实验研究[J].中国中医基础医学杂志,1997,3(3):28.
86.刘建秋.瓜蒌薤白半夏汤对肺动脉高压氧自由基的影响[J].中医药学报,1997,15(2):55.
87.贝伟剑,赵一.瓜蒌薤白汤的药理作用[J].中国医药学报,1989,4(5):341-343.
88.李向红.薤白的提取物抗抗氧化作用研究[J].中药材,1994,17(11):34.
89.张炳填,李鑫辉.栝蒌薤白半夏汤对急性心肌缺血大鼠血管内皮细胞保护作用的实验研究[J].新中医,2007,39(3):104-106.
90.刘萍,章怡,张静生.中药复方对损伤人血管内皮细胞表达细胞间黏附分子的影响[J].中国临床康复,2006,10(27):54-57.
91.郭书文,王国华.瓜蒌薤自半夏汤制剂对缺氧性肺动脉高压血NO、PAF的影响[J].北京中医药大 学学报,2001,24(2):37-38.
92.徐建民.加味瓜蒌薤白汤对心肌缺血动物模型的实验研究[J].湖北中医杂志[J].2007,29(4):8-9.
93.边秀娟.加味瓜蒌薤白半夏汤对哮喘小鼠血清IL-5的影响[J].河南中医学院学报,2007,22(133):18-20.
94.郑创华,陈汉锐,连乐粲,等.加味瓜蒌薤白汤不同提取液的抗凝和耐缺氧作用[J].中医药学刊,2006,24(6):1122.
95.宋建平,李瑞琴,李伟,等.瓜蒌薤白汤对肺纤维化大鼠肺组织中转化生长因子β1表达的影响[J].北京中医药大学学报,2005,28(2):40-43.
96.胡作为,周电光,周燕萍,等.中药止哮平喘方抗哮喘豚鼠模型气道炎症的实验研究[J].广州中医药大学学报,2004,21(1):40-43.
2.Hoeper MM,Mayer E,Simonneau Get al.Chronic thromboembolic pulmonary hypertension [J].Circulation,2006,113:2 011.
3.COOL CD,STEWART JS,WERAHERA P,et al.Three dimensional reconstruction of pulmonary arteries in plexiform pulmonary hypertension using cell specific markers:evidence for adynamic and heterogeneous process of pulmonary endothelial cell growth[J].A m J Pat hol,1999,155:411-419.
4.Wright JL,Levy RD,Churg A.Pulmonary hypertension in chronic obstructive pulmonary disease:current theories of pathogenesis and their implications for treatment[J].Thorax,2005,60(7):605-609.
5.Arao T,Takabatake N,Sata M,et al.In vivo evidence of endothelial injury in chronic obstructive pulmonary disease by lung scintigraphic assessment of(123) Imetaiodobenzylguanidine.[J].J Nucl Med,2003,44(11):1747-1754.
6.Bagatcheva NV,Garcia JG,Verin AD.Molecular mechanism ofthrombin-induced endothelial cell permeability[J].Biochemistry,2002,67:75-84.
7.陈瑞芬,周光德,曹文军,等.野百合碱诱导实验性肺动脉高压病理形态观察[J].电子显微学报,2002,21(1):1-4.
8.席思川,车东媛,张婉蓉,等.缺氧性肺动脉高压与肺腺泡内动脉构型重组[J].中国循环杂志,1995,(5):287-290.
9.王良兴,陈少贤,徐正介,等.低氧高二氧化碳环境下肺动脉内皮结构变化与肺动脉高压的关系[J].心肺血管病杂志,2000,19(3):220-223.
10.陈小客,邹霞英,辛达临.缺氧性肺动脉高压大鼠肺血管内皮细胞的变化[J].中国病理生理杂志,1996,12(6):608-611.
11.Voelkel NF,Cool CD.Pulmonary vascular involvement in chronic obstructive pulmonary disease[J].Eur Respir J Suppl,2003,46(suppl):28s-32s.
12.Furchgott RF,Zawadzki JV.The obligatory role of endothelial cells in the relation of arterial smooth muscle by acetylcheoling[J].Nature,1980,288(5789):373-376.
13.Voelkel NF,Cool C.Pathology of pulmonary hypertension[J].Cardiol Clin,2004,22:343-351.
14.Taraseviciene-Stewart L,Kasahare Y,Alger L,et al.Inhibition of the VEGF receptor 2 combined with chronic hypoxia causes cell death-dependent pulmonary endothelial cell proliferation and severe pulmonary hypertension[J].FASEB J,2001,15;427-438.
15.Hoshikawa Y,Ono S,Suzuki S,et al.Generation of oxidative stresss contributes to the development of pulmonary hypertension induced by hypoxia[J].J Appl Physionl,2001,90(4):1299-1306.
16.杨洁,易静,汤雪明.氧化修饰在调控细胞凋亡信号转导中的作用[J].细胞生物学杂志, 2005,27:121-126.
17.毛伟平,许峰.细胞凋亡过程中的信号转导[J].环境与职业医学,2004,21(1):62-65.
18.朱晓东.凋亡通路及其调控研究进展[J].国外医学,生理、病理科学与临床分册,2002,22(3):248-251.
19.Adams JM,Cory S.Life or death decisions by the Bcl-2 protein family[J].Trends Biochem Sci,2001,26(1):61-66.
20.KUWANA T,MACKEY M R,PERKINS G,et al.Bid,bax,and lipids cooperate to form supra molecular openings in the outer mitochondrial membrane[J].Cell,2002,111(3):331-342.
21.PETIT P X,ZAMZAMI N,VAYSSIERE J L,et al.Implication ofmitochondria in apoptosis[J].Mol Cell Biochem,1997,174(1-2):185-188.
22.NEWMEYER D D,FERGUSON-MILLER S.Mitochondria:Releasing power for life and unleashing machineries of death[J].Ce11,2003,112(4):481-490.
23.RAO RV,HERMEL E,CASTRO-OBREGON S,et al.Coupling endoplasmic reticulum stress to the cell death program:mechanism ofcaspase activation[J].J Biol Chem,2001,276(36):33869-33874.
24.Wallach D,Vorfolmeev EE,Malinin NL,et al.Tumor necrosis factor receptor and fas signaling mechanisms[J].Annu Rev Immunol,1999,17:331-367.
25.Karin M,Lin A.NF-kappa B at the crossroads of life and death.Nat Immunol.2002,3(3):221-227.
26.Michiels C,Minet E,Mottet D,et al.Regulation of gene expression by oxygen:NF-kappaB and HIF-1,two extremes[J].Free Radio Biol Med,2002,33(9):1231-1242.
27.Wang CY,Guttridge DC,Mayo MW,et al.NF-kappa B induces expression of the Bcl-2 homologue Al/Bfl-1 to preferentially suppress chemotherapy-induced apoptosis[J].Mol Cell Biol,1999,19:5923-5929.
28.Wang CY,Mayo MW,Komeluk RG,et al.NF-kappa B antiapoptosis:induction of TRAF1 and TRAF2 and c-IAP1 and c-IAP2 to suppress Caspase-8 activation[J].Science,1998,281:1680-1683.
29.Susin SA,Zamzami N,Castedo M,et al.The central executioner of apoptosis:multiple connections between protease activation and mitochondrial in Fas/APO-1/CD95-and ceramide-induced apoptosis [J].J Exp Med,1997,186:25-37.
30.王海燕,王来栓.细胞凋亡通路研究进展[J].国外医学生理病理科学与临床分册,2003,23(5):490-492.
31.Nakamura K,Bossy Wetzel E,Bums K,et al.Changes in endoplasmic reticulum luminal environment affect cell sensitivity to apoptosis[J].J Cell Biol,2000,150(4):731-740.
32.Rao RV,Hermel E,Castro-Obregon S,et al.Coupling endoplasmic reticulum stress to the cell death program.Mechanism of caspase activation[J].J Biol Chem,2001,276(36):33869-33874.
33.Pinton P,Ferrari D,Papizzi Z,et al.The Ca~(2+) concentration of the endoplasmic reticulum is a key determinant of ceramide-induced apoptosis:significance for the molecular mechanism of Bcl-2 action [J]. EMBo J,2001,20:2690-2701.
34. McConkey DJ , Nutt LK. Calciumflux measurements in apoptosis [J ]. Methods Cell Biol, 2001,66(2):229-246.
35. Nutt LK, Pataer A, Pahler J, et al. Bax and Bak promote apoptosis by modulating endoplasmic reticular and mitochondrial Ca~(2+) stores [J ]. J Biol Chem , 2002,277(11):9219-9225.
36. Breckenridge DG, Stojanovic M, Marcellus RC , et al. Caspase cleavage product of BAP31 induces mitochondrial fission through endoplasmic reticulum calcium signals , enhancing cytochrome release to the cytosol [J ]. J Cell Biol, 2003,160(7):l 115-1127.
37. Fujimoto K, Matsuzawa Y, Yamaguchi S. Benefits of oxygen on exercise performance and pulmonary hemodynamics in patients with COPD with mild hypoxemia[J ]. Chest ,2002 ,122 :457—463.
38. Franz IW, Van Der Meyden J, Schaupp S , et al. The effect of amlodipine on exercise—induced pulmonary hypertension and right heart function in patients with chronic obstructive pulmonary disease [J]. Z Kardiol ,2002,91 :833—839.
39. Borst MM ,Leschke M , Koing U ,et al. Repetitive hemodilution in chronic obstructive pulmonary disease and pulmonary hypertension :effects on pulmonary hemodynamics , gas exchange,and exercise capacity [J]. Respiration ,1999 ,66 :225—232.
40. Weg IL , Rossoff L , McKeon K. Development of pulmonary hypertension after lung volume reduction surgery [J]. Am J Respir Crit Care Med ,1999 ,159 :552-556.
41. Oswald—Mammosser M, Kessler R, Massard G, et al. Effect of lung volume reduction surgery on gas exchange and pulmonary hemodynamics at rest and during exercise [J]. Am J Respir Crit Care Med,1998 ,158:1020-1025.
42. Zhao YD, Courtman DW, Deng Y, et al. Rescue of monocrotaline induced pulmonary arterial hypertension using bone marrow-derived endothelial-like progenitor cells: efficacy of combined cell and eNOS gene therapy in established disease [J]. Circ Res, 2005.96:442-450.
43. Nagaya N, Kangawa K, Kanda M, et al. Hybrid cell-gene therapy for pulmonary hypertension based on phagocyteosing action of endpthelial progenitor cells [J]. Circulation, 2003,108:889-895.
44. KENTARO H, JUN F , YOSHIKO M, et al. Bone marrow derived cells cont ribute to pulmonary vascular remodeling in hypoxia-Induced pulmonary hypertension [J ]. Chest, 2005,127:1793-1798.
45. KIMIO S , YU TA KA K, MA KOTO N , et al. Important role of endogenous erythropoietin system in recruitment of endothelial progenitor cells in hypoxia-induced pulmonary hypertension in mice [J].Circulation, 2006 ,113 :1442-1450.
46. NAGA YA N , KANGAWA K, KANDA M , et al. Hybrid cell-gene therapy for pulmonary hypertension based on phagocytosing action of endothelial progenitor cells [J].Ciculation,2003,108 :889-895.
47.周仲瑛.周仲瑛临床经验辑要[M].北京:中国医药科技出版社,1998.1(1):36.
48.李素云,吴其标.曹世宏教授论治慢性阻塞性肺疾病经验选粹[J].中医药学刊.2002,20(1):28.
49.奚肇庆,刘学华,冷报浪,等.通阳宣痹法治疗慢性阻塞性肺病急性发作期临床研究[J].中国中医急症,2005,14(11):1025-1027.
50.奚肇庆,蒋荫,居文政,等.复方薤白胶囊治疗慢性阻塞性肺病36例临床与实验研究[J].中医杂志,2000,4(41):218-220.
51.奚肇庆.通阳宣痹法治疗支气管哮喘急性发作期的临床研究[J].江苏中医,1999,20(2):14-15.
52.冷报浪,奚肇庆,曹世宏,等.复方薤白胶囊对慢性支气管炎大鼠血小板活化因子及一氧化氮的影响[J].现代中西医结合杂志,2006,15(10):1293-1299.
53.孙菊光,奚肇庆,姜静,等.复方薤白胶囊对野百合碱诱导的肺动脉高压大鼠病理的影响[J].南京中医药大学学报(自然科学版),2006,22(3):173-176.
54.孙菊光,奚肇庆,姜静,等.复方薤白胶囊对野百合碱诱导肺动脉高压大鼠SOD活性、MDA、HYP含量的影响[J].实用中医药杂志,2005,21(12):711-713.
55.孙菊光,奚肇庆,姜静,等.复方薤白胶囊对野百合碱诱导肺动脉高压大鼠ET、TNF α含量的影响[J].东南大学学报(医学版),2006,25(5):337-340.
56.姜静,尚宁,奚肇庆,等.复方薤白胶囊对野百合碱诱导肺动脉高压大鼠血浆中花生四烯酸代谢产物的影响[J].中华结核和呼吸杂志,2005,28(8):534-536.
57.奚肇庆,姜静,孙菊光,等.复方薤白胶囊对野百合碱诱导大鼠肺动脉高压的抑制作用[J].中医杂志,2006,47(10):781-783.
58.王剑蓉,朱云华.复方薤白胶囊治疗实验性慢性支气管炎的病理观察[J].南京中医药大学学报(自然科学版),2002,18(4):224-225.
59.陈丽萍,白旭东,于国梁.高分辨率超声观察薤自制剂对高脂血症的治疗作用[J].中国老年学杂志,2002,22(5):182.
60.谭可安.薤白的临床疗效观察[J].白求恩医科大学学报,1989.15(2):211.
61.陈滴.薤白对家兔体内前列腺素El的影响[J].白求恩医科大学学报,1989.15(1):91.
62.孟庆国,朱庆磊,邓淑娥,等.薤白水提物对羟自由基的清除作用[J].潍坊医学院学报,1998,20(1):66.
63.沈树人.“薤白桂皮汤”治疗毛细支气管炎合并肠炎50例[J].中西医结合实用临床急症,1997,10(4):462-463.
64.李向红.薤白的提取物抗氧化作用研究[J].中药材,1994,17(11):34.
65.吴洪元.薤白的炮制研究[J].中药材,1995,18(4):192.
66.吴波,陈思维,王伟敏,等.瓜蒌薤白白酒汤提取物抗心肌缺血缺氧及最佳处方的筛选[J].中草药,2000,31(11):844.
67.张卿,高尔,侯琦,等.薤白挥发油对S180荷瘤小鼠免疫功能的影响[J].潍坊医学院学报,2002,24(2):94.
68.阴健,郭力弓主编.中药现代研究与临床应用[M].北京:学院出版社,1993,260-263.
69.吴波,王敏伟,陈思维,等.瓜蒌提取物对离体家兔胸主动脉条收缩的影响[J].沈阳药科大学学报,1999,16(1):24-27.
70.贝伟剑.瓜蒌薤白汤的药理作用[J].中国医药学报,1989,4(5):341.
71.邵春丽,王世久,王进,等.瓜蒌皮抗缺氧作用的研究[J].沈阳药科大学学报,1998,15(1):38-40.
72.向邦平,金善炜,傅桂香,等.瓜蒌蛋白Ⅱ.瓜萎蛋白部分化学结构的初步测定[J].化学学报,1998.56:302-303.
73.李仪奎.中药药理学[M].北京:中国中医药出版社,1992:157.
74.沈雅琴,张明发.半夏的镇痛、抗溃疡和抗血栓作用的形成[J].中国生化药物杂志,1998,19(3):141.
75.刘继林.水半夏与半夏部分药理作用的对比研究[J].成都中医药学院学报,1989,12(2):41.
76.洪行球.半夏降血脂作用研究[J].浙江中医学院学报,1995,19(2):28-29.
77.王光明,周蓉.半夏的中药药理研究进展[J].中医药导报,2007,13(2):97-99.
78.Marin Nero JA,Maciel BC,Secches AL,et al.Cardiovascular effects of berberine in patients with severe congestive heart failure[J].Clin Car diol,1988,11(4):253-260.
79.周祖玉,孙爱民,徐建国,等.黄连素对离体灌流心脏的能量保存作用[J].华西医科大学学报,2002,33(3):431-433.
80.Zeng XH,Zeng XJ,Li YY.Efficacy and safety of berberine for congestive heart failure secondary to ischemic or idiopathic dilated cardiomyopathy[J].AmJ Cardiol,2003,92(2):173-176.
81.袁拯忠,朱陵群,庞鹤等.黄连解毒汤有效成分对缺氧/复氧时脑微血管内皮细胞的保护作用[J].中国中药杂志,2007,32(3):249-252.
82.崔学军.黄连及其有效成分的药理研究进展[J].中国药师,2006,9(5):469-470.
83.王海燕,李玉清,王启会.黄连素抗肿瘤作用研究进展[J].中成药,2007,29(10):1500-1502.
84.陈彬,张琪伟.瓜蒌薤白药对大鼠心功能及血流变的影响[J].南京中医药大学学报,1996,12(2):26.
85.徐长胜.瓜蒌薤白方加味止咳祛痰作用的实验研究[J].中国中医基础医学杂志,1997,3(3):28.
86.刘建秋.瓜蒌薤白半夏汤对肺动脉高压氧自由基的影响[J].中医药学报,1997,15(2):55.
87.贝伟剑,赵一.瓜蒌薤白汤的药理作用[J].中国医药学报,1989,4(5):341-343.
88.李向红.薤白的提取物抗抗氧化作用研究[J].中药材,1994,17(11):34.
89.张炳填,李鑫辉.栝蒌薤白半夏汤对急性心肌缺血大鼠血管内皮细胞保护作用的实验研究[J].新中医,2007,39(3):104-106.
90.刘萍,章怡,张静生.中药复方对损伤人血管内皮细胞表达细胞间黏附分子的影响[J].中国临床康复,2006,10(27):54-57.
91.郭书文,王国华.瓜蒌薤自半夏汤制剂对缺氧性肺动脉高压血NO、PAF的影响[J].北京中医药大 学学报,2001,24(2):37-38.
92.徐建民.加味瓜蒌薤白汤对心肌缺血动物模型的实验研究[J].湖北中医杂志[J].2007,29(4):8-9.
93.边秀娟.加味瓜蒌薤白半夏汤对哮喘小鼠血清IL-5的影响[J].河南中医学院学报,2007,22(133):18-20.
94.郑创华,陈汉锐,连乐粲,等.加味瓜蒌薤白汤不同提取液的抗凝和耐缺氧作用[J].中医药学刊,2006,24(6):1122.
95.宋建平,李瑞琴,李伟,等.瓜蒌薤白汤对肺纤维化大鼠肺组织中转化生长因子β1表达的影响[J].北京中医药大学学报,2005,28(2):40-43.
96.胡作为,周电光,周燕萍,等.中药止哮平喘方抗哮喘豚鼠模型气道炎症的实验研究[J].广州中医药大学学报,2004,21(1):40-43.