重用龟版治疗肾阴虚再生障碍性贫血的临床分析与实验研究
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摘要
研究目的
再生障碍性贫血(简称再障)是由化学、物理、生物因素、药物及不明原因引起的骨髓干细胞、造血微环境损伤,红骨髓被脂肪代替,全血细胞减少的一组综合征。造血微环境异常是再障发病的重要因素之一,骨髓间充质干细胞(MSC)作为骨髓造血微环境中的一种重要细胞成分,近年来成为干细胞研究领域的一大热点。再障属中医学“虚劳”、“血证”、“亡血”范畴。中医认为“肾主骨生髓、藏精”、“精血互生”,而再障发病的关键在于“肾虚、髓枯”,因此“补肾填精”被认为是治疗再障的基本法则。导师丘和明教授对再障进行了多年的研究,提出肾虚血瘀是再障根本病机,认为再障的好转是按肾阴虚向肾阳虚的规律归转的。中药复方治疗的效果亦显示出对肾阳虚者疗效显著,而对肾阴虚者较差,体现了中医理论“阳虚易治,阴虚难调”客观实际。
本课题临床研究部分,旨在观察重用龟版补肾填精治疗肾阴虚型再障的临床疗效;实验研究部分,以5-氟尿嘧啶与白消安合用诱导建立大鼠再障模型,观察龟版对再障模型大鼠的治疗作用、病理形态变化及MSC体外增殖活性的影响,探讨其作用机制,为龟版治疗再障的临床运用提供实验依据。
研究方法
临床研究:采用自身前后对照试验,观察重用龟版治疗肾阴虚型再障的临床疗效。方法为,将符合西医再障诊断标准及中医肾阴虚证候诊断标准的12例再障病人纳入研究,服用重用龟版的补肾填精方2个疗程,观察治疗前后临床证候积分、症状改善、外周血网织红细胞、白细胞、血红蛋白含量及血小板的变化,进行临床疗效分析。
实验研究:50只清洁级雌性SD大鼠按1:4随机化原则随机分为正常对照组10只,造模组40只。造模组以5-FU与白消安合用诱导建立再障大鼠模型。将造模成功并存活的32只大鼠随机分为4组:模型对照组、龟版低剂量组、龟版中剂量组、龟版高剂量组,除模型对照组外,其它3组分别灌服不同剂量的龟版水煎液3周。观察所有实验动物的血常规动态变化、骨髓有核细胞数、病理形态、骨髓MSC体外培养及活性检测。
临床研究成果和结论
1.临床观察12例病人,随防8～22个月,缓解4例,明显进步6例,无效2例,总有效率83.33%。
2.12例病人经重用龟版补肾填精治疗2个疗程后,临床证候学积分明显下降,症状明显改善,外周血网织红细胞、白细胞计数、血红蛋白含量比治疗前明显升高,血小板亦略有上升,病人输血及血小板的例次明显减少。
结论:重用龟版补肾填精治疗肾阴虚型再障取得良好近期疗效。
Objective of the research:Aplastic anemia is due to chemical, physical and biological factors. Medication and other unknown elements are also to blame for this disease. It causes the damage of haemopoietic microenvironment of stem cells in the marrow. The red bone marrow is replaced by fat and whole blood red cell is on the decrease. Abnormality of haemopoietic microenvironment is an important factor of aplastic anemia. As an essential cell component, MSC has become the focus of stem cell research. Aplastic anemia, in the eyes of traditional Chinese medicine, falls into the category of "deficiency of fatigue", "blood disease" and "massive haemorrhage". Traditional Chinese medicine thinks "the kidney is in charge of the bone and gives birth to marrow and stores essential energy" and "Qi and blood are syngenetic" . The key of aplastic anemia lies in the deficiency of kidney Qi and the dryness of marrow. As a result, making up for the deficiency of kidney Qi and enriching the essential energy is considered the fundamental principle in curing aplastic anemia. My academic advisor, Professor Qiu Heming has done years of research in the field of aplastic anemia. He thinks that pathological mechanism of aplastic anemia is the deficiency of kidney Qi and blood stasis. He considers that the recovery of aplastic anemia follows the rule of changing from deficiency of Yin in the kidney to deficiency of Yang. Traditional Chinese medicinal compound administration also has a positive effect on the deficiency of Yang in kidney, but no effect on the deficiency of Yin in kidney. This embodies the deep-rooted idea in traditional Chinese medicine that "Deficiency of Yang heals easily but the deficiency of Yin is hard to deal with".The clinical research part of this project aims to observe the clinical effect of tortoise plastron as an enrichment of kidney on aplastic anemia which falls into the category of deficiency of Yin. By establishing model of aplastic anemia in rats, the experimental research part of the project observes the clinical effective influence of tortoise plastron on aplastic anemic rats, their pathological change, MSC and extracorporeal proliferation activity, explores the working mechanism, and provides
experimental evidence for the clinical application of tortoise plastron as a cure for aplastic anemia. Research method:Clinical study: A comparison of subjects themselves before and after the experiment is conducted. The method of observing the clinical effectiveness of tortoise plastron on aplastic anemia with deficiency of Yin in kidney is as follows: 12 patients are involved in the study who, in the eyes of western medicine, are aplastic anemic patients but in the eyes of traditional Chinese medicine, have symptoms of deficiency of Yin in kidney. After two courses of treatment by taking tortoise plastron as an enrichment of essential energy in kidney, a change before and after the experiment is observed in the clinical symptom index, improvement of symptoms, peripheral blood reticulated corpuscles, content of hemoglobin and platelet. A clinical effectiveness analysis is conducted.In the experiment, 50 clean female SD rats are classified into two groups at random. 10 rats are in a control group while 40 are in the modeling group. By a consecutive use of 5-FU and busulfan, these 40 rats are established as an aplastic anemic model group. 8 rats die as a result of the drugs and the surviving 32 rats are divided into 4 groups at random. 1: model control group; 2: low dose of tortoise plastron; 3: medium dose of tortoise plastron; 4: high dose of tortoise plastron. Then rats are fed different doses of tortoise plastron for 3 weeks, except the model control group. The dynamic change in the routine blood test in all the experimental animals is observed, together with the number of nucleated cells, pathological state, MSC, extracorporeal cultivation and the test of activity. Clinical research result and conclusion:1. Of the 12 patients, with 8 to 22 months of follow-up visits taken into consideration, there are 4 people showing signs of alleviation, 6 marked improvement, 2 with no effect at all. The general effectiveness is 83.33%.2. After two courses of high-dose in the 12 patients, the index of clinical symptoms have fallen. There is a marked improvement in the symptoms, reticulated corpuscles and the white blood cell count. The content of hemoglobin has shown a marked increase compared with the state before the experiment. The number of platelets has risen and the number of both blood transfusions and platelet transfusions has decreased.Conclusion: The near-term effectiveness of high-dose of tortoise plastron as an enrichment of kidney with the deficiency of Yin is good. Experimental result and conclusion:1. By using first 5-FU to gather stem cells and then busulfan to destroy the accumulated stem cells, a combination of these two medicines succeeds in establishing a model in which rats suffer aplastic anemia because of a failure of stem cells.
再生障碍性贫血(简称再障)是由化学、物理、生物因素、药物及不明原因引起的骨髓干细胞、造血微环境损伤,红骨髓被脂肪代替,全血细胞减少的一组综合征。造血微环境异常是再障发病的重要因素之一,骨髓间充质干细胞(MSC)作为骨髓造血微环境中的一种重要细胞成分,近年来成为干细胞研究领域的一大热点。再障属中医学“虚劳”、“血证”、“亡血”范畴。中医认为“肾主骨生髓、藏精”、“精血互生”,而再障发病的关键在于“肾虚、髓枯”,因此“补肾填精”被认为是治疗再障的基本法则。导师丘和明教授对再障进行了多年的研究,提出肾虚血瘀是再障根本病机,认为再障的好转是按肾阴虚向肾阳虚的规律归转的。中药复方治疗的效果亦显示出对肾阳虚者疗效显著,而对肾阴虚者较差,体现了中医理论“阳虚易治,阴虚难调”客观实际。
本课题临床研究部分,旨在观察重用龟版补肾填精治疗肾阴虚型再障的临床疗效;实验研究部分,以5-氟尿嘧啶与白消安合用诱导建立大鼠再障模型,观察龟版对再障模型大鼠的治疗作用、病理形态变化及MSC体外增殖活性的影响,探讨其作用机制,为龟版治疗再障的临床运用提供实验依据。
研究方法
临床研究:采用自身前后对照试验,观察重用龟版治疗肾阴虚型再障的临床疗效。方法为,将符合西医再障诊断标准及中医肾阴虚证候诊断标准的12例再障病人纳入研究,服用重用龟版的补肾填精方2个疗程,观察治疗前后临床证候积分、症状改善、外周血网织红细胞、白细胞、血红蛋白含量及血小板的变化,进行临床疗效分析。
实验研究:50只清洁级雌性SD大鼠按1:4随机化原则随机分为正常对照组10只,造模组40只。造模组以5-FU与白消安合用诱导建立再障大鼠模型。将造模成功并存活的32只大鼠随机分为4组:模型对照组、龟版低剂量组、龟版中剂量组、龟版高剂量组,除模型对照组外,其它3组分别灌服不同剂量的龟版水煎液3周。观察所有实验动物的血常规动态变化、骨髓有核细胞数、病理形态、骨髓MSC体外培养及活性检测。
临床研究成果和结论
1.临床观察12例病人,随防8～22个月,缓解4例,明显进步6例,无效2例,总有效率83.33%。
2.12例病人经重用龟版补肾填精治疗2个疗程后,临床证候学积分明显下降,症状明显改善,外周血网织红细胞、白细胞计数、血红蛋白含量比治疗前明显升高,血小板亦略有上升,病人输血及血小板的例次明显减少。
结论:重用龟版补肾填精治疗肾阴虚型再障取得良好近期疗效。
Objective of the research:Aplastic anemia is due to chemical, physical and biological factors. Medication and other unknown elements are also to blame for this disease. It causes the damage of haemopoietic microenvironment of stem cells in the marrow. The red bone marrow is replaced by fat and whole blood red cell is on the decrease. Abnormality of haemopoietic microenvironment is an important factor of aplastic anemia. As an essential cell component, MSC has become the focus of stem cell research. Aplastic anemia, in the eyes of traditional Chinese medicine, falls into the category of "deficiency of fatigue", "blood disease" and "massive haemorrhage". Traditional Chinese medicine thinks "the kidney is in charge of the bone and gives birth to marrow and stores essential energy" and "Qi and blood are syngenetic" . The key of aplastic anemia lies in the deficiency of kidney Qi and the dryness of marrow. As a result, making up for the deficiency of kidney Qi and enriching the essential energy is considered the fundamental principle in curing aplastic anemia. My academic advisor, Professor Qiu Heming has done years of research in the field of aplastic anemia. He thinks that pathological mechanism of aplastic anemia is the deficiency of kidney Qi and blood stasis. He considers that the recovery of aplastic anemia follows the rule of changing from deficiency of Yin in the kidney to deficiency of Yang. Traditional Chinese medicinal compound administration also has a positive effect on the deficiency of Yang in kidney, but no effect on the deficiency of Yin in kidney. This embodies the deep-rooted idea in traditional Chinese medicine that "Deficiency of Yang heals easily but the deficiency of Yin is hard to deal with".The clinical research part of this project aims to observe the clinical effect of tortoise plastron as an enrichment of kidney on aplastic anemia which falls into the category of deficiency of Yin. By establishing model of aplastic anemia in rats, the experimental research part of the project observes the clinical effective influence of tortoise plastron on aplastic anemic rats, their pathological change, MSC and extracorporeal proliferation activity, explores the working mechanism, and provides
experimental evidence for the clinical application of tortoise plastron as a cure for aplastic anemia. Research method:Clinical study: A comparison of subjects themselves before and after the experiment is conducted. The method of observing the clinical effectiveness of tortoise plastron on aplastic anemia with deficiency of Yin in kidney is as follows: 12 patients are involved in the study who, in the eyes of western medicine, are aplastic anemic patients but in the eyes of traditional Chinese medicine, have symptoms of deficiency of Yin in kidney. After two courses of treatment by taking tortoise plastron as an enrichment of essential energy in kidney, a change before and after the experiment is observed in the clinical symptom index, improvement of symptoms, peripheral blood reticulated corpuscles, content of hemoglobin and platelet. A clinical effectiveness analysis is conducted.In the experiment, 50 clean female SD rats are classified into two groups at random. 10 rats are in a control group while 40 are in the modeling group. By a consecutive use of 5-FU and busulfan, these 40 rats are established as an aplastic anemic model group. 8 rats die as a result of the drugs and the surviving 32 rats are divided into 4 groups at random. 1: model control group; 2: low dose of tortoise plastron; 3: medium dose of tortoise plastron; 4: high dose of tortoise plastron. Then rats are fed different doses of tortoise plastron for 3 weeks, except the model control group. The dynamic change in the routine blood test in all the experimental animals is observed, together with the number of nucleated cells, pathological state, MSC, extracorporeal cultivation and the test of activity. Clinical research result and conclusion:1. Of the 12 patients, with 8 to 22 months of follow-up visits taken into consideration, there are 4 people showing signs of alleviation, 6 marked improvement, 2 with no effect at all. The general effectiveness is 83.33%.2. After two courses of high-dose in the 12 patients, the index of clinical symptoms have fallen. There is a marked improvement in the symptoms, reticulated corpuscles and the white blood cell count. The content of hemoglobin has shown a marked increase compared with the state before the experiment. The number of platelets has risen and the number of both blood transfusions and platelet transfusions has decreased.Conclusion: The near-term effectiveness of high-dose of tortoise plastron as an enrichment of kidney with the deficiency of Yin is good. Experimental result and conclusion:1. By using first 5-FU to gather stem cells and then busulfan to destroy the accumulated stem cells, a combination of these two medicines succeeds in establishing a model in which rats suffer aplastic anemia because of a failure of stem cells.
引文
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