泻肺逐饮汤对恶性胸腔积液大鼠TGF-β1、AQP1、CTLA-4和PFP的影响
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摘要
目的：通过观察泻肺逐饮汤对恶性胸腔积液大鼠模型胸水和胸膜组织中TGF-β1、AQP1、CTLA-4.PFP及生存期的影响,探讨泻肺逐饮汤对恶性胸腔积液的作用机制及疗效评价。
方法：本实验将100只大鼠随机分为空白对照组(A)、模型组(B)、中药组(C)、力尔凡组(D)、中药加力尔凡组(E)5组,每组20只除空白对照组外,其余四组大鼠以1X108/l的艾氏腹水瘤细胞悬液胸腔内注射造模。从造模次日起,空白对照组、模型组予生理盐水灌胃7天；中药组、中药加力尔凡组给予泻肺逐饮汤灌胃,每日一次,连续7天；力尔凡组、中药加力尔凡组予力尔凡腹腔注射,隔日1次。造模后第8天取材,每组取12只鼠检测相关指标,剩余8只做生存期观察。测量各组大鼠胸水量的变化；光镜下观察胸膜组织和胸水中的细胞形态的变化；用免疫组化法测定胸水和胸膜组织中TGF-β1、AQP1的表达,用RT-PCR法测定胸水和胸膜组织中的CTLA-4和PFP的表达。
结果：
1.三个治疗组大鼠中位生存时间及生命延长率均较模型组延长(p<0.05),中药组与中药加力尔凡组较力尔凡组延长(p<0.05)。
2.治疗组大鼠的胸水量较模型组少(p<0.05),而中药组和中药加力尔凡组差异明显(p<0.01)
3.光镜下观察,模型组胸水涂片中,可见到大量癌细胞聚集成团,形态不规则。模型组胸水沉渣HE染色,可见大量癌细胞,形态不规则,胞浆比例严重失调。模型组大鼠的胸膜明显增厚、严重破坏,可见大量癌细胞核增大浓染,部分可见成团出现。各治疗组较模型组程度减轻。
4.模型组大鼠胸水中TGF-β1含量高于各治疗组,与中药组、力尔凡组比较有差异(p<0.05),与中药加力尔凡组比较差异显著(p<0.01)；力尔凡组、中药组与中药加力尔凡组比较无差异(p>0.05)。模型组大鼠胸膜组织中TGF-β1含量高于各治疗组,与中药组和中药加力尔凡组比较有显著差异(p<0.01)；中药组的含量低于力尔凡组(p<0.05)；中药加力尔凡组的含量明显低于中药组和力尔凡组(p<0.01)。
5.模型组大鼠胸水中AQP1含量高于各治疗组,与中药组、力尔凡组比较有差异(p<0.05),与中药加力尔凡组比较差异显著(p<0.01)；力尔凡组、中药组与中药加力尔凡组比较无差异(p>0.05)。模型组大鼠胸膜组织中AQP1含量明显高于各治疗组,比较有显著差异(p<0.01)；中药加力尔凡组的含量明显低于中药组和力尔凡组(p<0.01)。
6.模型组胸水和胸膜组织中CTLA-4mRNA表达较高,与中药组、力尔凡组比较有差异(p<0.05),与中药加力尔凡组比较差异显著(p<0.01)；中药组和力尔凡组胸膜中CTLA-4mRNA表达高于中药加力尔凡组(p<0.05)；力尔凡组高于中药组(p<0.05)。
7.模型组大鼠胸水中PFPmRNA表达较低,与三个治疗组比较有明显差异(p<0.01)；力尔凡组明显低于中药组、中药加力尔凡组(p<0.01)。模型组大鼠胸膜组织中PFPmRNA表达较低,与中药组和力尔凡组比较有差异(p<0.05),与中药加力尔凡组比较有差异显著(p<0.01)；中药加力尔凡组高于中药组和力尔凡组(p<0.05)。
结论：
1.泻肺逐饮汤可以延长恶性胸腔积液大鼠的生存期。
2.泻肺逐饮汤可以减少恶性胸腔积液大鼠胸水量。
3.泻肺逐饮汤可以降低恶性胸腔积液大鼠胸水和胸膜组织中TGF-β1、AQP1、CTLA-4的含量。
4.泻肺逐饮汤可以增加恶性胸腔积液大鼠胸水和胸膜组织中穿孔素的表达,从而治疗恶性胸腔积液
Objective:This experiment is to approach the mechanisms of xiefeizhuyin Decoction to MPE and the evaluation the curative effects of that, through the observation on the level of TGF betal,AQP1,CTLA-4and PFP in the pleural effusion and tissue on rat models of Malignant pleural effusion(MPE) and the survival rates of rats
Method:100rats were divided into five groups randomly:blank group(A group),model group(A group), Chinese materia mediea group (C group), Mannatide injection group(D group) and Chinese materia mediea with Mannatide injection group(E group),20rats in each group.To set up80MPE rat models except blank group, were injected with EAC(Ehrliehs ascites carcinoma) in their pleural cavity.We observe the survival rates of rat models, detected the level of TGF betal and AQP1by immunohistoehemisty and the expression of CTLA-4and PFP by RT-PCR.
Result:
1. Compared with model group, median survival time and life extension rate in the C、D、E groups is longer(p<0.05), while that of C group and E group is longer than D group (p<0.05).
2. Compared with model group, the pleural effusion volume of D groups (p<0.05) decreased,that of C and E group decreased Significantly (p<0.01).
3. Observed under the light microscope, model group effusion smear,it is visible to see a large number of cancer cells to clump together with irregular shape. In the model group chest water sediment HE staining, there were a large number of cancer cells,with irregular shape and cytoplasmic serious imbalance. The pleural of the model group is thicken and serious damaged, there is a great deal of cancer cells with nuclei increased dense and clusters appear partially. About the above indexes, the treatment groups reduce the degree of that compared with model group.
4. The TGF betal content on pleural effusion of model group is higher than that of C and D groups (p<0.05),much higher than that of the E group(p<0.01). The TGF betal content on pleural tissue of model group is significantly higher than that of C、Dand E groups (p<0.01); D group is higher than that of the C group(p<0.05); C and D groups are much higher than that of the E group(p<0.01).
5. The AQP1content on pleural effusion of model group is higher than that of C and D groups (p<0.05),much higher than that of the E group(p<0.01). C and D group are higher than E group on that (p<0.05). The AQP1content on pleural effusion and pleural tissue of model group is higher than that of C and D groups (p<0.05),much higher than that of the E group(p<0.01). D group is higher than E group on that (p<0.05).
6. The CTLA-4content on pleural effusion and pleural tissue of model group is higher than that of C and D groups (p<0.05),much higher than that of the E group(p<0.01). D group is higher than E group on that (p<0.05). The CTLA-4content on pleural tissue of E group is lower than C and D group (p<0.05). The CTLA-4content on pleural tissue of C and D group is lower than D group (p<0.05).
7. The PFP content on pleural effusion of model group is much lower than that of each treatment groups (p<0.01),while D group is lower than that of the C and E group (p<0.01). The PFP content on pleural tissue of model group is lower than that of C and D groups (p<0.05),much higher than that of the E group (p<0.01). The PFP content on pleural tissue of E group is higher than C and D groups(p<0.01).
Conclusion:
1. Xiefei Zhuyin decoction can extend the malignant pleural effusion rats on the survival period.
2. Xiefei Zhuyin decoction reduce the pleural effusion on the malignant pleural effusion rats
3. Xiefei Zhuyin decoction reduce the TGF beta, AQP1, CTLA-4 content on pleural effusion and pleural tissue of malignant pleural effusion rat.
4. Xiefei Zhuyin decoction increase the PFP content on pleural effusion and pleural tissue of malignant pleural effusion rat, enhance the PFP in the activity, thus for the treatment of malignant pleural effusion.
方法：本实验将100只大鼠随机分为空白对照组(A)、模型组(B)、中药组(C)、力尔凡组(D)、中药加力尔凡组(E)5组,每组20只除空白对照组外,其余四组大鼠以1X108/l的艾氏腹水瘤细胞悬液胸腔内注射造模。从造模次日起,空白对照组、模型组予生理盐水灌胃7天；中药组、中药加力尔凡组给予泻肺逐饮汤灌胃,每日一次,连续7天；力尔凡组、中药加力尔凡组予力尔凡腹腔注射,隔日1次。造模后第8天取材,每组取12只鼠检测相关指标,剩余8只做生存期观察。测量各组大鼠胸水量的变化；光镜下观察胸膜组织和胸水中的细胞形态的变化；用免疫组化法测定胸水和胸膜组织中TGF-β1、AQP1的表达,用RT-PCR法测定胸水和胸膜组织中的CTLA-4和PFP的表达。
结果：
1.三个治疗组大鼠中位生存时间及生命延长率均较模型组延长(p<0.05),中药组与中药加力尔凡组较力尔凡组延长(p<0.05)。
2.治疗组大鼠的胸水量较模型组少(p<0.05),而中药组和中药加力尔凡组差异明显(p<0.01)
3.光镜下观察,模型组胸水涂片中,可见到大量癌细胞聚集成团,形态不规则。模型组胸水沉渣HE染色,可见大量癌细胞,形态不规则,胞浆比例严重失调。模型组大鼠的胸膜明显增厚、严重破坏,可见大量癌细胞核增大浓染,部分可见成团出现。各治疗组较模型组程度减轻。
4.模型组大鼠胸水中TGF-β1含量高于各治疗组,与中药组、力尔凡组比较有差异(p<0.05),与中药加力尔凡组比较差异显著(p<0.01)；力尔凡组、中药组与中药加力尔凡组比较无差异(p>0.05)。模型组大鼠胸膜组织中TGF-β1含量高于各治疗组,与中药组和中药加力尔凡组比较有显著差异(p<0.01)；中药组的含量低于力尔凡组(p<0.05)；中药加力尔凡组的含量明显低于中药组和力尔凡组(p<0.01)。
5.模型组大鼠胸水中AQP1含量高于各治疗组,与中药组、力尔凡组比较有差异(p<0.05),与中药加力尔凡组比较差异显著(p<0.01)；力尔凡组、中药组与中药加力尔凡组比较无差异(p>0.05)。模型组大鼠胸膜组织中AQP1含量明显高于各治疗组,比较有显著差异(p<0.01)；中药加力尔凡组的含量明显低于中药组和力尔凡组(p<0.01)。
6.模型组胸水和胸膜组织中CTLA-4mRNA表达较高,与中药组、力尔凡组比较有差异(p<0.05),与中药加力尔凡组比较差异显著(p<0.01)；中药组和力尔凡组胸膜中CTLA-4mRNA表达高于中药加力尔凡组(p<0.05)；力尔凡组高于中药组(p<0.05)。
7.模型组大鼠胸水中PFPmRNA表达较低,与三个治疗组比较有明显差异(p<0.01)；力尔凡组明显低于中药组、中药加力尔凡组(p<0.01)。模型组大鼠胸膜组织中PFPmRNA表达较低,与中药组和力尔凡组比较有差异(p<0.05),与中药加力尔凡组比较有差异显著(p<0.01)；中药加力尔凡组高于中药组和力尔凡组(p<0.05)。
结论：
1.泻肺逐饮汤可以延长恶性胸腔积液大鼠的生存期。
2.泻肺逐饮汤可以减少恶性胸腔积液大鼠胸水量。
3.泻肺逐饮汤可以降低恶性胸腔积液大鼠胸水和胸膜组织中TGF-β1、AQP1、CTLA-4的含量。
4.泻肺逐饮汤可以增加恶性胸腔积液大鼠胸水和胸膜组织中穿孔素的表达,从而治疗恶性胸腔积液
Objective:This experiment is to approach the mechanisms of xiefeizhuyin Decoction to MPE and the evaluation the curative effects of that, through the observation on the level of TGF betal,AQP1,CTLA-4and PFP in the pleural effusion and tissue on rat models of Malignant pleural effusion(MPE) and the survival rates of rats
Method:100rats were divided into five groups randomly:blank group(A group),model group(A group), Chinese materia mediea group (C group), Mannatide injection group(D group) and Chinese materia mediea with Mannatide injection group(E group),20rats in each group.To set up80MPE rat models except blank group, were injected with EAC(Ehrliehs ascites carcinoma) in their pleural cavity.We observe the survival rates of rat models, detected the level of TGF betal and AQP1by immunohistoehemisty and the expression of CTLA-4and PFP by RT-PCR.
Result:
1. Compared with model group, median survival time and life extension rate in the C、D、E groups is longer(p<0.05), while that of C group and E group is longer than D group (p<0.05).
2. Compared with model group, the pleural effusion volume of D groups (p<0.05) decreased,that of C and E group decreased Significantly (p<0.01).
3. Observed under the light microscope, model group effusion smear,it is visible to see a large number of cancer cells to clump together with irregular shape. In the model group chest water sediment HE staining, there were a large number of cancer cells,with irregular shape and cytoplasmic serious imbalance. The pleural of the model group is thicken and serious damaged, there is a great deal of cancer cells with nuclei increased dense and clusters appear partially. About the above indexes, the treatment groups reduce the degree of that compared with model group.
4. The TGF betal content on pleural effusion of model group is higher than that of C and D groups (p<0.05),much higher than that of the E group(p<0.01). The TGF betal content on pleural tissue of model group is significantly higher than that of C、Dand E groups (p<0.01); D group is higher than that of the C group(p<0.05); C and D groups are much higher than that of the E group(p<0.01).
5. The AQP1content on pleural effusion of model group is higher than that of C and D groups (p<0.05),much higher than that of the E group(p<0.01). C and D group are higher than E group on that (p<0.05). The AQP1content on pleural effusion and pleural tissue of model group is higher than that of C and D groups (p<0.05),much higher than that of the E group(p<0.01). D group is higher than E group on that (p<0.05).
6. The CTLA-4content on pleural effusion and pleural tissue of model group is higher than that of C and D groups (p<0.05),much higher than that of the E group(p<0.01). D group is higher than E group on that (p<0.05). The CTLA-4content on pleural tissue of E group is lower than C and D group (p<0.05). The CTLA-4content on pleural tissue of C and D group is lower than D group (p<0.05).
7. The PFP content on pleural effusion of model group is much lower than that of each treatment groups (p<0.01),while D group is lower than that of the C and E group (p<0.01). The PFP content on pleural tissue of model group is lower than that of C and D groups (p<0.05),much higher than that of the E group (p<0.01). The PFP content on pleural tissue of E group is higher than C and D groups(p<0.01).
Conclusion:
1. Xiefei Zhuyin decoction can extend the malignant pleural effusion rats on the survival period.
2. Xiefei Zhuyin decoction reduce the pleural effusion on the malignant pleural effusion rats
3. Xiefei Zhuyin decoction reduce the TGF beta, AQP1, CTLA-4 content on pleural effusion and pleural tissue of malignant pleural effusion rat.
4. Xiefei Zhuyin decoction increase the PFP content on pleural effusion and pleural tissue of malignant pleural effusion rat, enhance the PFP in the activity, thus for the treatment of malignant pleural effusion.
引文
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