补肾通络法对输卵管炎性阻塞模型大鼠作用机理的理论及实验研究
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摘要
目的:输卵管炎性阻塞好发于育龄期妇女,常导致不孕。本研究以中医学基本理论为依据、结合现代中医学研究成果及我们的临床工作经验,探讨输卵管炎性阻塞的中医机理,并通过实验研究观察补肾通络法对FTIO大鼠组织形态学、血液流变学、细胞因子(IL-2、IL-6、TNF-α)、ICAM-1蛋白、Bcl-2蛋白的影响,探讨其作用机制,以指导临床治疗。
方法:以SD雌性大鼠为受试对象,用苯酚糊剂输卵管注入法复制输卵管炎性阻塞性模型,将60只大鼠随机分为正常组、模型组、妇炎康组、及补肾通络方高、中、低剂量组。采用HE染色观察输卵管组织形态学变化、电镜观察输卵管组织超微结构、全自动血粘度仪检测血液流变学指标、ELISA法检测血清中IL-2.、IL-6和TNF-a含量、应用免疫组化检测输卵管上皮细胞ICAM-1和BcL-2蛋白表达。
结果:1.模型组大鼠输卵管组织光、电镜下观察符合慢性炎症病理改变,治疗后有明显改善。2.模型组大鼠血液流变学各项指标均高于正常组,符合“血瘀证”改变,治疗后高剂量组基本恢复正常。3.FTIO模型大鼠血清IL-2含量明显低于正常,IL-6、TNF-α含量明显高于正常,补肾通络方能对上述细胞因子有调节作用,且疗效优于妇炎康。4.FTIO大鼠输卵管组织中的ICAM-1蛋白表达明显高于正常组,补肾通络方能降低其表达。5.FTIO模型组大鼠输卵管上皮Bcl-2蛋白表达明显较增多,治疗后各组大鼠输卵管上皮Bcl-2蛋白表达不同程度减少,以高剂量组明显。
结论:1.用苯酚糊剂输卵管注入法可以成功复制输卵管炎性阻塞性模型。2.补肾通络法能治疗输卵管炎性阻塞,一方面可能是中药能直接抗炎、改善微循环、促进炎症细胞及其他增生细胞凋亡、促进受损组织修复,另一方面可能是中药能作用于免疫系统,通过对细胞因子、粘附分子等的调节而改善其失衡状态,从而增强其抗病能力、自我修复能力。
Objective:The fallopian tube inflammatory obstructive(FTIO)is one of the common diseases andgynaecological refractory,the disease occurs in Childbearing years woman,it is also one of the major causeses of infertility.In this study,we discuss the TCM theory of FTIO.In this study,rats Salpingitis phenol paste model of obstruction,by observing the effect of the Kidney-Tonifying and Meridian-Dredging Method on FTIO rats morphology,hemorheology,cytokines(IL-2,IL-6,TNF-a),ICAM-1 and Bcl-2 protein,and to explore the mechanism for the clinical treatment of obstructive Salpingitis result of infertility and provide theoretical And experimental basis.
Methods:To 8-12 months old and weighing 180-220 grams of SD is not pregnant female rats to test targets,with phenol paste into law copy Salpingitis tubal occlusion of the model.60 rats were divided into six groups,namely,Tonify the kidney,high-dose group(the high-dose group), Tonify the kidney dose group(in the dose group),Tonify the kidney, low-dose group(Low-dose group),Fu Yah Kang group,model group,the normal group(sham group).Gavages for 20 days after delivery,pick eyeball blood after rats were killed,quickly anatomy,observe the shape and color of tubal and the surrounding tissue adhesion there,there effusion;Remove bilateral tubal,rapidly add to 10 percent neutral formaldehyde fixative or 2.5%glutaraldehyde in stationary liquid fixed, sliced,use light microscopy to observe the Organization of tubal morphological changes,use the electron microscopy to observe tubal ultrastructure;use the Automatic blood viscosity to measure hemorheology indicators;ELISA method in serum IL-2.,IL-6 and TNF-a content; immunohistochemical detection of epithelial cells tubal ICAM-1 and BcL-2 protein expression.
Results:1.Eye In model group fallopian tube and surrounding tissue are different degrees of adhesion,the lumen of uneven thickness,hardens, the color of pale,some visible tubal thickening,water and even pyonephrosis.2.The results showed that,after treatment,tubal appearance,light,electricity under observation with the model groups have markedly improved,Tonify the kidney law that could reduce the chronic inflammation,thus Salpingitis treatment of obstructive role.3. Hemorheology model group of indicators including blood viscosity,red blood cell aggregation index,the index of rigid red blood cells, hematocrit were significantly higher than the normal group.This study shows that the law tonify the kidney FTIO rats Hemorheology of the indicators have improved significantly,and some indicators of the improvement is better than the control group.4.To sum up,tonify the kidney role of law in Salpingitis obstruction of rats,the model caused by IL-2 lower,IL-6 and TNF-a increased the abnormal increase significantly corrected,and the IL-2 regulation effective than the control group.5.The tnotify the kidney can inhibit inflammation caused by abnormal increase ICAM-1 and thus play its anti-inflammatory effect. 6.The tnotify the kidney may be by regulating apoptosis suppressor gene Bcl-2 protein expression,thereby inhibiting pathogenic microbes to stimulate induced apoptosis,to control inflammation,relieve inflammation caused by tissue injury purposes.
Conclusion:1.Phenol paste tubal injection can copy the successful model of Salpingitis obstruction.2.The Kidney-Tonifying and Meridian-Dredging Method to the treatment of obstructive Salpingitis,on the one hand may be able to direct anti-inflammatory medicine,improve microcirculation and inhibit apoptosis,and promote tissue repair damage to the other medicine may be able to Role in the immune system,through the cytokines,adhesion molecules,such as the regulation and improve their state of imbalance,thereby enhancing their ability to resistance and self-repair capacity.
方法:以SD雌性大鼠为受试对象,用苯酚糊剂输卵管注入法复制输卵管炎性阻塞性模型,将60只大鼠随机分为正常组、模型组、妇炎康组、及补肾通络方高、中、低剂量组。采用HE染色观察输卵管组织形态学变化、电镜观察输卵管组织超微结构、全自动血粘度仪检测血液流变学指标、ELISA法检测血清中IL-2.、IL-6和TNF-a含量、应用免疫组化检测输卵管上皮细胞ICAM-1和BcL-2蛋白表达。
结果:1.模型组大鼠输卵管组织光、电镜下观察符合慢性炎症病理改变,治疗后有明显改善。2.模型组大鼠血液流变学各项指标均高于正常组,符合“血瘀证”改变,治疗后高剂量组基本恢复正常。3.FTIO模型大鼠血清IL-2含量明显低于正常,IL-6、TNF-α含量明显高于正常,补肾通络方能对上述细胞因子有调节作用,且疗效优于妇炎康。4.FTIO大鼠输卵管组织中的ICAM-1蛋白表达明显高于正常组,补肾通络方能降低其表达。5.FTIO模型组大鼠输卵管上皮Bcl-2蛋白表达明显较增多,治疗后各组大鼠输卵管上皮Bcl-2蛋白表达不同程度减少,以高剂量组明显。
结论:1.用苯酚糊剂输卵管注入法可以成功复制输卵管炎性阻塞性模型。2.补肾通络法能治疗输卵管炎性阻塞,一方面可能是中药能直接抗炎、改善微循环、促进炎症细胞及其他增生细胞凋亡、促进受损组织修复,另一方面可能是中药能作用于免疫系统,通过对细胞因子、粘附分子等的调节而改善其失衡状态,从而增强其抗病能力、自我修复能力。
Objective:The fallopian tube inflammatory obstructive(FTIO)is one of the common diseases andgynaecological refractory,the disease occurs in Childbearing years woman,it is also one of the major causeses of infertility.In this study,we discuss the TCM theory of FTIO.In this study,rats Salpingitis phenol paste model of obstruction,by observing the effect of the Kidney-Tonifying and Meridian-Dredging Method on FTIO rats morphology,hemorheology,cytokines(IL-2,IL-6,TNF-a),ICAM-1 and Bcl-2 protein,and to explore the mechanism for the clinical treatment of obstructive Salpingitis result of infertility and provide theoretical And experimental basis.
Methods:To 8-12 months old and weighing 180-220 grams of SD is not pregnant female rats to test targets,with phenol paste into law copy Salpingitis tubal occlusion of the model.60 rats were divided into six groups,namely,Tonify the kidney,high-dose group(the high-dose group), Tonify the kidney dose group(in the dose group),Tonify the kidney, low-dose group(Low-dose group),Fu Yah Kang group,model group,the normal group(sham group).Gavages for 20 days after delivery,pick eyeball blood after rats were killed,quickly anatomy,observe the shape and color of tubal and the surrounding tissue adhesion there,there effusion;Remove bilateral tubal,rapidly add to 10 percent neutral formaldehyde fixative or 2.5%glutaraldehyde in stationary liquid fixed, sliced,use light microscopy to observe the Organization of tubal morphological changes,use the electron microscopy to observe tubal ultrastructure;use the Automatic blood viscosity to measure hemorheology indicators;ELISA method in serum IL-2.,IL-6 and TNF-a content; immunohistochemical detection of epithelial cells tubal ICAM-1 and BcL-2 protein expression.
Results:1.Eye In model group fallopian tube and surrounding tissue are different degrees of adhesion,the lumen of uneven thickness,hardens, the color of pale,some visible tubal thickening,water and even pyonephrosis.2.The results showed that,after treatment,tubal appearance,light,electricity under observation with the model groups have markedly improved,Tonify the kidney law that could reduce the chronic inflammation,thus Salpingitis treatment of obstructive role.3. Hemorheology model group of indicators including blood viscosity,red blood cell aggregation index,the index of rigid red blood cells, hematocrit were significantly higher than the normal group.This study shows that the law tonify the kidney FTIO rats Hemorheology of the indicators have improved significantly,and some indicators of the improvement is better than the control group.4.To sum up,tonify the kidney role of law in Salpingitis obstruction of rats,the model caused by IL-2 lower,IL-6 and TNF-a increased the abnormal increase significantly corrected,and the IL-2 regulation effective than the control group.5.The tnotify the kidney can inhibit inflammation caused by abnormal increase ICAM-1 and thus play its anti-inflammatory effect. 6.The tnotify the kidney may be by regulating apoptosis suppressor gene Bcl-2 protein expression,thereby inhibiting pathogenic microbes to stimulate induced apoptosis,to control inflammation,relieve inflammation caused by tissue injury purposes.
Conclusion:1.Phenol paste tubal injection can copy the successful model of Salpingitis obstruction.2.The Kidney-Tonifying and Meridian-Dredging Method to the treatment of obstructive Salpingitis,on the one hand may be able to direct anti-inflammatory medicine,improve microcirculation and inhibit apoptosis,and promote tissue repair damage to the other medicine may be able to Role in the immune system,through the cytokines,adhesion molecules,such as the regulation and improve their state of imbalance,thereby enhancing their ability to resistance and self-repair capacity.
引文
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1.陈鏌鎁.苯酚胶浆剂注入输卵管绝育的病理过程及其成败原因的探讨[J].中华妇产科杂志,1979,1 4(2):84
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