降糖三黄片对高糖诱导的心肌细胞TGF-β1调控的影响及临床研究
|
摘要
第一部分文献研究
糖尿病(diabetes mellitus, DM)是由遗传和环境因素共同引起的一组以糖代谢紊乱为主要表现的临床综合征。全世界的糖尿病患病率迅速增加,发展中国家尤为明显,已经成为临床上的主要内分泌代谢病。据国际糖尿病联盟(IDF)估计,截至2025年,我国糖尿病患者人数将达到5930万[1]。糖尿病心脏病(Diabetic Cardiopathy, DC),是指糖尿病合并或伴发的心脏病变,此病名于1979年由Ledet首先提出,很快得到世界同行专家认可。该病包括冠状动脉粥样硬化性心脏病、糖尿病性心肌病、微血管病变和植物神经功能紊乱所致的心律失常等[2]。这几种病变常共同存在,互为因果,相互促进,形成恶性循环。大动脉的粥样硬化(颈动脉、主动脉、股动脉)增加了心肌梗塞、中风及截肢的危[3]。糖尿病心肌病(Diabetic cardiomyopathy)首先由Rubler等在解剖患有充血性心力衰竭的糖尿病患者尸体时提出的[4],其病理表现主要为心脏微血管病变、心肌代谢紊乱和心肌纤维化等所致的心肌广泛结构异常,最终引起左心室肥厚、舒张期和(或)收缩期功能障碍的一种疾病状态[5]。对于心肌细胞的探索,目前已成为糖尿病心肌病的研究热点,利用培养的心肌细胞,探索非血液动力学因素所致的心肌肥厚的调节,研究心肌细胞的生物力学、凋亡、受体下调、缺血和处理、信号通路,对作用于心脏的新药进行筛选并对其安全性进行评价,以及从培养的心肌细胞中提取有价值的生物因子,同药物诱导干细胞分化而成的心肌细胞作阳性对照等。
中医中药对糖尿病心脏病的治疗越来越受到学术界的重视,中药处方以其显著的疗效,低廉的成本,较少的副作用深受患者青睐。以熊曼琪教授为首的研究课题组在上世纪80年代围绕气阴两虚、瘀血阻滞为病机,以《伤寒论》中桃核承气汤为底方,研制成院内制剂——降糖三黄片,在临床及机理研究方面都取得了较大的进展,本实验在诸研究基础上进一步探讨降糖三黄片在分子细胞水平的生物学机制。
第二部分实验研究
目的：观察降糖三黄片对高糖诱导下心肌细胞凋亡率影响,观察心肌细胞TGF-β1表达,进一步从信号通路角度探讨降糖三黄片的作用机理。
方法：
1.提取含药血清。普通级雄性新西兰兔8只,随机分为正常组2只,降糖三黄片低剂量组2只、中剂量组2只、高剂量组2只,普通饲料适应性喂养3天,降糖三黄片低剂量组,予0.75g／kg灌胃；中剂量组予1.5g／kg灌胃；高剂量组予3g／kg灌胃,正常组灌入等量蒸馏水。灌服1周后,禁食12h,腹主动脉采血,提取含药血清。-70℃冰箱冻存。
2.心肌细胞培养及鉴定。Wistar雄性出生3天乳鼠,取心脏组织,0.25%EDTA消化过夜,15h后,予等量全培中止消化,予胶原消化酶Ⅱ消化,将心肌细胞与红细胞充分分离,将心肌细胞按5×105/mL移至12孔细胞培养板中。入37℃二氧化碳培养箱培养。24h后,可见心肌细胞自行跳动,且生长成片,予Desmin免疫组化鉴定,1:100Desmin组胞浆中有棕黄色颗粒的阳性细胞占总细胞数85%左右,磷酸缓冲液(PBS)阴性对照组未见阳性细胞,则成功培养心肌细胞。
3.流式细胞仪检测细胞凋亡。将700ul含葡萄糖(33mmol/L)的全培分别加入降糖三黄片低、中、高剂量组及模型组,再分别予不同剂量降糖三黄片血清300ul,模型组及高糖组加入全培,干预后入二氧化碳培养箱培养48h后,流式细胞仪检测各组心肌细胞凋亡率。
4.ELISA检测心肌细胞TGF-β1含量。将同检测细胞凋亡干预的心肌细胞上清液离心后,取上清,用ELISA法检测各组的OD值,并根据标准曲线,计算各组的TGF-β1含量。
结果：流式细胞仪观察心肌细胞凋亡率,正常组与模型组相比较具有显著差异(p<0.05),降糖三黄片低、中、高组间比较具有显著性差异(p<0.05),且其心肌细胞凋亡率随剂量增加而减少,降糖三黄片组与模型组具统计学差异(p<0.05),说明降糖三黄片可以抑制高糖诱导的心肌细胞凋亡,且随药物剂量增大,其抑制凋亡率程度越明显; ELISA检测心肌细胞TGF-β1示降糖三黄片低剂量组与高剂量组无统计学差异(p>0.05),中剂量组与高剂量、低剂量组有统计学差异(p<0.05),同时低剂量组、高剂量组与模型组相比较无统计学意义(p>0.05),中剂量组与模型组相比较具有统计学意义(p<0.05),说明降糖三黄片低、高剂量对于乳鼠心肌细胞TGF-β1的表达并无显著影响,而中剂量降糖三黄片组可显著下调TGF-β1的表达。
结论：1.中药降糖三黄片可有效改善高糖诱导的心肌细胞凋亡,随浓度增加,抑制凋亡率程度越强。2.中药降糖三黄片中剂量可以显著下调高糖诱导的心肌细胞TGF-β1的表达,其机制可能与上调系膜细胞的GLUT1表达,继而激活蛋白激酶C (PKC)氨基己糖通路有关。
第三部分临床研究
纳入观察病例82例,均明确诊断为糖尿病心脏病。采用随机分组表将患者分为降糖三黄片组和基础治疗组,经1周治疗,对照组患者治疗前后TGF-β1含量比较,经t检验,没有统计学意义(P值为0.94),即对照组患者经治疗后TGF-β1表达下降不明显。对照组患者治疗前后TGF-β1含量比较,经t检验,没有统计学意义(P值为0.94),即对照组患者经治疗后TGF-β1表达下降不明显。降糖三黄片组患者治疗前后TGF-β1含量比较,经t检验,有统计学意义(P值为0.00),即降糖三黄片组患者经治疗后TGF-β1表达下降。治疗前降糖三黄片组与基础治疗组TGF-β1含量比较,经t检验,两组均无统计学意义(P值为0.09),即两组具可比性。
对患者性别,年龄,体重指数,糖尿病病程,血压,血脂,糖化血红蛋白,证型,血压,胰岛素2h释放进行多元回归线性统计分析,发现诸混杂因素中,只有证型对服用降糖三黄片治疗患者血清TGF-β1的下调有影响,即在服用降糖三黄片的人群中,辨证为气阴两虚兼有瘀血证的患者TGF-β1的表达下调更明显。
Diabetes (diabetes mellitus, DM) is a group which causes together by the heredity and the environmental factor take the sugar metabolic disorder as the main performance clinical syndrome. The world diabetes prevalence rate increases rapidly, the developing country is especially obvious, already became clinical on the main endocrine metabolic disease. According to the international diabetes alliance (IDF) estimated that up to2025, our country diabetic population will achieve59,300,000t. Diabetes heart disease (Diabetic Cardiopathy, DC), refers to the heart pathological change which diabetes merge or the partner send, this sickness name first proposed in1979by Ledet, obtains the world colleague expert to approve very quickly. This sickness including coronal atherosclerosis heart disease, diabetes myocardiopathy, capillaries pathological change and nervus autonomici function disorder result arrythmia and so on[3]. These kind of sickness coexist abnormal, the cause and effect relationship, promotes, to form the vicious circle mutually. Aorta's gruel type hardening (carotid artery, aorta, femoral artery) increased myocardial infarction, the stroke and the amputation danger. Diabetes myocardiopathy (Diabetic cardiomyopathy) first by Rubler and so on when the dissection contracts the congested heart failure the diabetic corpse proposes, its pathology performance mainly for heart result and so on capillaries pathological change, cardiac muscle metabolic disorder and cardiac muscle fibrosis cardiac muscle widespread textural anomalies, causes the left ventricle to be plump finally, relaxing period and (or) systole function barrier one kind of disease condition. Regarding myocardial cell's exploration, has become diabetes myocardiopathy's research hot spot at present, the use raise myocardial cell, explores the non-blood dynamics factor result the cardiac muscle plump adjustment, research myocardial cell's biological mechanics, perishes, the acceptor to decline, the ischemia and processing, the signal passage weakly, to affects carries on screening in heart's new medicine and carries on the appraisal to its security, as well as withdraws the valuable biotic factor from the raise myocardial cell, becomes the myocardial cell with the medicine induction stem cell differentiation to make the masculine comparison and so on.
The Chinese medicine traditional Chinese medicine receives the academic circle more and more diabetes heart disease's treatment the value, the traditional Chinese medicine prescription by its remarkable curative effect, the inexpensive cost, the few side effect depth is favored the patient. Take Professor Xiong Man Qi as head's research topic-based group in the1980s regarding was mad that cloudy two empty, the ecchymosis hindered for the pathogenesis, the peach seed received by "Typhoid" was mad that Tang Weide the side, developed Cheng Yuannei the preparation-to fall the sugar three yellow pieces, has made the big progress in cl inical and the mechanism research aspect, this experiment further discussed in various research foundation falls sugar three Huang Pian in the molecular cell level biology mechanism. Second part of experimental study goal:The observation falls sugar three Huang Pian to the high sugar induction under the myocardial cell weak perishing rate influence, the observation myocardial cell TGF-β1expression, further falls the sugar three Huang Pian action mechanism from the signal passage angle discussion. Method:1. the extraction contains the medicine blood serum. Ordinary level male New Zealand rabbit8, divide into normal group2stochastically, falls sugar three Huang Piandi monitoring team2, monitoring team2, high monitoring team2, the ordinary feed compatibility feeds for3days, falls sugar three Huang Piandi the monitoring team, gives0.75g/kg to fill the stomach; The monitoring team gives1.5g/kg to fill the stomach; The high monitoring team gives3g/kg to fill the stomach, the normal group pours into the isometric distilled water. Fills the clothing1week later, fasts12h, the abdomen aorta picks the blood, the extraction contains the medicine blood serum.-70℃the refrigerator saves frozen.2. myocardial cell raise and appraisal. The Wistar male gender is born3day of young mouse, takes the heart organization, the0.25%EDTA digestion over night, after15h, gives the isometric to cultivate stops digesting, gives the collagen digestive ferment ⅡThe digestion, separates fully the myocardial cell and the red blood cell, according to5X105/mL moves to12cell culture board the myocardial cell. Enters37℃the carbon dioxide incubator raise. after24h, obviously the myocardial cell beats voluntarily, and grows Large expanse, gives the Desmin immunity group appraisal, in the1:100Desmin group cytoblastema has the yellowish brown pellet masculine cell to occupy the total cell several about85%, the phosphoric acid buffer solution (PBS) the negative control group has not seen the masculine cell, then success raise myocardial cell.3. the class type cell meter examination cell perishes weakly.700ul including glucose (30mmol/L) all cultivates joins separately falls the sugar three yellow piece of low, middle and high monitoring teams and the model group, gives the different dosage to fall sugar three yellow piece of blood serum300ul separately again, the model group and Gao Tangzu joins all cultivates, after the intervention, after entering the carbon dioxide incubator raises48h, flows the type cell meter to examine each group of myocardial cell weak perishing rate.4. ELISA examination myocardial cell TGF-β1content. Perishes weakly after the examination cell the intervention myocardial cell supernate offcenter, is accepted after passing an examination clear, examines each group with the ELISA law the OD value, and according to the specification curve, calculates each group TGF-β1content. Finally:Flows type cell meter observation myocardial cell weak perishing rate, the normal group and the model group compare have the remarkable difference (p<0.05), falls the sugar during three yellow piece of low, middle and high groups the comparison to have the significance difference (p<0.05), and his/her myocardial cell weak perishing rate increases along with the dosage reduces, falls sugar three Huang Pianzu and the model group statistics difference (p<0.05), explained that falls the sugar three yellow pieces to be possible to suppress the high sugar induction the myocardial cell to perish weakly, and increases along with the medicine dosage, it suppresses the weak perishing rate degree to be more obvious; The ELISA examination myocardial cell TGF-β1shows falls sugar three Huang Piandi the monitoring team and the high monitoring team non-statistics difference (p>0.05), the monitoring team and the high dose, the low monitoring team have statistics difference (p<0.05), simultaneously lowers the monitoring team, the high monitoring team and the model group compares the non-statistics significance (p>0.05), the monitoring team and the model group compare have statistics significance (p<0.05), explained that falls sugar three Huang Piandi, the high dose and not obviously affects regarding the young mouse myocardial cell TGF-β1expression, but the dosage falls sugar three Huang Pianzu to be possible obviously to decline TGF-β1expression. Conclusion:1. the traditional Chinese medicine falls sugar three Huang Pianke to improve the high sugar induction the myocardial cell to perish weakly effectively, increases along with the density, suppresses the weak perishing rate degree to be stronger.2. the traditional Chinese medicine falls in sugar three Huang Pian the dosage to be possible remarkable under to raise the sugar induction the myocardial cell TGF-β1expression, its mechanism is possible and the upward is the membrane cell's GLUT1expression, subsequently activates protein kinase C (PKC) the amino sugar circuit to concern.
糖尿病(diabetes mellitus, DM)是由遗传和环境因素共同引起的一组以糖代谢紊乱为主要表现的临床综合征。全世界的糖尿病患病率迅速增加,发展中国家尤为明显,已经成为临床上的主要内分泌代谢病。据国际糖尿病联盟(IDF)估计,截至2025年,我国糖尿病患者人数将达到5930万[1]。糖尿病心脏病(Diabetic Cardiopathy, DC),是指糖尿病合并或伴发的心脏病变,此病名于1979年由Ledet首先提出,很快得到世界同行专家认可。该病包括冠状动脉粥样硬化性心脏病、糖尿病性心肌病、微血管病变和植物神经功能紊乱所致的心律失常等[2]。这几种病变常共同存在,互为因果,相互促进,形成恶性循环。大动脉的粥样硬化(颈动脉、主动脉、股动脉)增加了心肌梗塞、中风及截肢的危[3]。糖尿病心肌病(Diabetic cardiomyopathy)首先由Rubler等在解剖患有充血性心力衰竭的糖尿病患者尸体时提出的[4],其病理表现主要为心脏微血管病变、心肌代谢紊乱和心肌纤维化等所致的心肌广泛结构异常,最终引起左心室肥厚、舒张期和(或)收缩期功能障碍的一种疾病状态[5]。对于心肌细胞的探索,目前已成为糖尿病心肌病的研究热点,利用培养的心肌细胞,探索非血液动力学因素所致的心肌肥厚的调节,研究心肌细胞的生物力学、凋亡、受体下调、缺血和处理、信号通路,对作用于心脏的新药进行筛选并对其安全性进行评价,以及从培养的心肌细胞中提取有价值的生物因子,同药物诱导干细胞分化而成的心肌细胞作阳性对照等。
中医中药对糖尿病心脏病的治疗越来越受到学术界的重视,中药处方以其显著的疗效,低廉的成本,较少的副作用深受患者青睐。以熊曼琪教授为首的研究课题组在上世纪80年代围绕气阴两虚、瘀血阻滞为病机,以《伤寒论》中桃核承气汤为底方,研制成院内制剂——降糖三黄片,在临床及机理研究方面都取得了较大的进展,本实验在诸研究基础上进一步探讨降糖三黄片在分子细胞水平的生物学机制。
第二部分实验研究
目的：观察降糖三黄片对高糖诱导下心肌细胞凋亡率影响,观察心肌细胞TGF-β1表达,进一步从信号通路角度探讨降糖三黄片的作用机理。
方法：
1.提取含药血清。普通级雄性新西兰兔8只,随机分为正常组2只,降糖三黄片低剂量组2只、中剂量组2只、高剂量组2只,普通饲料适应性喂养3天,降糖三黄片低剂量组,予0.75g／kg灌胃；中剂量组予1.5g／kg灌胃；高剂量组予3g／kg灌胃,正常组灌入等量蒸馏水。灌服1周后,禁食12h,腹主动脉采血,提取含药血清。-70℃冰箱冻存。
2.心肌细胞培养及鉴定。Wistar雄性出生3天乳鼠,取心脏组织,0.25%EDTA消化过夜,15h后,予等量全培中止消化,予胶原消化酶Ⅱ消化,将心肌细胞与红细胞充分分离,将心肌细胞按5×105/mL移至12孔细胞培养板中。入37℃二氧化碳培养箱培养。24h后,可见心肌细胞自行跳动,且生长成片,予Desmin免疫组化鉴定,1:100Desmin组胞浆中有棕黄色颗粒的阳性细胞占总细胞数85%左右,磷酸缓冲液(PBS)阴性对照组未见阳性细胞,则成功培养心肌细胞。
3.流式细胞仪检测细胞凋亡。将700ul含葡萄糖(33mmol/L)的全培分别加入降糖三黄片低、中、高剂量组及模型组,再分别予不同剂量降糖三黄片血清300ul,模型组及高糖组加入全培,干预后入二氧化碳培养箱培养48h后,流式细胞仪检测各组心肌细胞凋亡率。
4.ELISA检测心肌细胞TGF-β1含量。将同检测细胞凋亡干预的心肌细胞上清液离心后,取上清,用ELISA法检测各组的OD值,并根据标准曲线,计算各组的TGF-β1含量。
结果：流式细胞仪观察心肌细胞凋亡率,正常组与模型组相比较具有显著差异(p<0.05),降糖三黄片低、中、高组间比较具有显著性差异(p<0.05),且其心肌细胞凋亡率随剂量增加而减少,降糖三黄片组与模型组具统计学差异(p<0.05),说明降糖三黄片可以抑制高糖诱导的心肌细胞凋亡,且随药物剂量增大,其抑制凋亡率程度越明显; ELISA检测心肌细胞TGF-β1示降糖三黄片低剂量组与高剂量组无统计学差异(p>0.05),中剂量组与高剂量、低剂量组有统计学差异(p<0.05),同时低剂量组、高剂量组与模型组相比较无统计学意义(p>0.05),中剂量组与模型组相比较具有统计学意义(p<0.05),说明降糖三黄片低、高剂量对于乳鼠心肌细胞TGF-β1的表达并无显著影响,而中剂量降糖三黄片组可显著下调TGF-β1的表达。
结论：1.中药降糖三黄片可有效改善高糖诱导的心肌细胞凋亡,随浓度增加,抑制凋亡率程度越强。2.中药降糖三黄片中剂量可以显著下调高糖诱导的心肌细胞TGF-β1的表达,其机制可能与上调系膜细胞的GLUT1表达,继而激活蛋白激酶C (PKC)氨基己糖通路有关。
第三部分临床研究
纳入观察病例82例,均明确诊断为糖尿病心脏病。采用随机分组表将患者分为降糖三黄片组和基础治疗组,经1周治疗,对照组患者治疗前后TGF-β1含量比较,经t检验,没有统计学意义(P值为0.94),即对照组患者经治疗后TGF-β1表达下降不明显。对照组患者治疗前后TGF-β1含量比较,经t检验,没有统计学意义(P值为0.94),即对照组患者经治疗后TGF-β1表达下降不明显。降糖三黄片组患者治疗前后TGF-β1含量比较,经t检验,有统计学意义(P值为0.00),即降糖三黄片组患者经治疗后TGF-β1表达下降。治疗前降糖三黄片组与基础治疗组TGF-β1含量比较,经t检验,两组均无统计学意义(P值为0.09),即两组具可比性。
对患者性别,年龄,体重指数,糖尿病病程,血压,血脂,糖化血红蛋白,证型,血压,胰岛素2h释放进行多元回归线性统计分析,发现诸混杂因素中,只有证型对服用降糖三黄片治疗患者血清TGF-β1的下调有影响,即在服用降糖三黄片的人群中,辨证为气阴两虚兼有瘀血证的患者TGF-β1的表达下调更明显。
Diabetes (diabetes mellitus, DM) is a group which causes together by the heredity and the environmental factor take the sugar metabolic disorder as the main performance clinical syndrome. The world diabetes prevalence rate increases rapidly, the developing country is especially obvious, already became clinical on the main endocrine metabolic disease. According to the international diabetes alliance (IDF) estimated that up to2025, our country diabetic population will achieve59,300,000t. Diabetes heart disease (Diabetic Cardiopathy, DC), refers to the heart pathological change which diabetes merge or the partner send, this sickness name first proposed in1979by Ledet, obtains the world colleague expert to approve very quickly. This sickness including coronal atherosclerosis heart disease, diabetes myocardiopathy, capillaries pathological change and nervus autonomici function disorder result arrythmia and so on[3]. These kind of sickness coexist abnormal, the cause and effect relationship, promotes, to form the vicious circle mutually. Aorta's gruel type hardening (carotid artery, aorta, femoral artery) increased myocardial infarction, the stroke and the amputation danger. Diabetes myocardiopathy (Diabetic cardiomyopathy) first by Rubler and so on when the dissection contracts the congested heart failure the diabetic corpse proposes, its pathology performance mainly for heart result and so on capillaries pathological change, cardiac muscle metabolic disorder and cardiac muscle fibrosis cardiac muscle widespread textural anomalies, causes the left ventricle to be plump finally, relaxing period and (or) systole function barrier one kind of disease condition. Regarding myocardial cell's exploration, has become diabetes myocardiopathy's research hot spot at present, the use raise myocardial cell, explores the non-blood dynamics factor result the cardiac muscle plump adjustment, research myocardial cell's biological mechanics, perishes, the acceptor to decline, the ischemia and processing, the signal passage weakly, to affects carries on screening in heart's new medicine and carries on the appraisal to its security, as well as withdraws the valuable biotic factor from the raise myocardial cell, becomes the myocardial cell with the medicine induction stem cell differentiation to make the masculine comparison and so on.
The Chinese medicine traditional Chinese medicine receives the academic circle more and more diabetes heart disease's treatment the value, the traditional Chinese medicine prescription by its remarkable curative effect, the inexpensive cost, the few side effect depth is favored the patient. Take Professor Xiong Man Qi as head's research topic-based group in the1980s regarding was mad that cloudy two empty, the ecchymosis hindered for the pathogenesis, the peach seed received by "Typhoid" was mad that Tang Weide the side, developed Cheng Yuannei the preparation-to fall the sugar three yellow pieces, has made the big progress in cl inical and the mechanism research aspect, this experiment further discussed in various research foundation falls sugar three Huang Pian in the molecular cell level biology mechanism. Second part of experimental study goal:The observation falls sugar three Huang Pian to the high sugar induction under the myocardial cell weak perishing rate influence, the observation myocardial cell TGF-β1expression, further falls the sugar three Huang Pian action mechanism from the signal passage angle discussion. Method:1. the extraction contains the medicine blood serum. Ordinary level male New Zealand rabbit8, divide into normal group2stochastically, falls sugar three Huang Piandi monitoring team2, monitoring team2, high monitoring team2, the ordinary feed compatibility feeds for3days, falls sugar three Huang Piandi the monitoring team, gives0.75g/kg to fill the stomach; The monitoring team gives1.5g/kg to fill the stomach; The high monitoring team gives3g/kg to fill the stomach, the normal group pours into the isometric distilled water. Fills the clothing1week later, fasts12h, the abdomen aorta picks the blood, the extraction contains the medicine blood serum.-70℃the refrigerator saves frozen.2. myocardial cell raise and appraisal. The Wistar male gender is born3day of young mouse, takes the heart organization, the0.25%EDTA digestion over night, after15h, gives the isometric to cultivate stops digesting, gives the collagen digestive ferment ⅡThe digestion, separates fully the myocardial cell and the red blood cell, according to5X105/mL moves to12cell culture board the myocardial cell. Enters37℃the carbon dioxide incubator raise. after24h, obviously the myocardial cell beats voluntarily, and grows Large expanse, gives the Desmin immunity group appraisal, in the1:100Desmin group cytoblastema has the yellowish brown pellet masculine cell to occupy the total cell several about85%, the phosphoric acid buffer solution (PBS) the negative control group has not seen the masculine cell, then success raise myocardial cell.3. the class type cell meter examination cell perishes weakly.700ul including glucose (30mmol/L) all cultivates joins separately falls the sugar three yellow piece of low, middle and high monitoring teams and the model group, gives the different dosage to fall sugar three yellow piece of blood serum300ul separately again, the model group and Gao Tangzu joins all cultivates, after the intervention, after entering the carbon dioxide incubator raises48h, flows the type cell meter to examine each group of myocardial cell weak perishing rate.4. ELISA examination myocardial cell TGF-β1content. Perishes weakly after the examination cell the intervention myocardial cell supernate offcenter, is accepted after passing an examination clear, examines each group with the ELISA law the OD value, and according to the specification curve, calculates each group TGF-β1content. Finally:Flows type cell meter observation myocardial cell weak perishing rate, the normal group and the model group compare have the remarkable difference (p<0.05), falls the sugar during three yellow piece of low, middle and high groups the comparison to have the significance difference (p<0.05), and his/her myocardial cell weak perishing rate increases along with the dosage reduces, falls sugar three Huang Pianzu and the model group statistics difference (p<0.05), explained that falls the sugar three yellow pieces to be possible to suppress the high sugar induction the myocardial cell to perish weakly, and increases along with the medicine dosage, it suppresses the weak perishing rate degree to be more obvious; The ELISA examination myocardial cell TGF-β1shows falls sugar three Huang Piandi the monitoring team and the high monitoring team non-statistics difference (p>0.05), the monitoring team and the high dose, the low monitoring team have statistics difference (p<0.05), simultaneously lowers the monitoring team, the high monitoring team and the model group compares the non-statistics significance (p>0.05), the monitoring team and the model group compare have statistics significance (p<0.05), explained that falls sugar three Huang Piandi, the high dose and not obviously affects regarding the young mouse myocardial cell TGF-β1expression, but the dosage falls sugar three Huang Pianzu to be possible obviously to decline TGF-β1expression. Conclusion:1. the traditional Chinese medicine falls sugar three Huang Pianke to improve the high sugar induction the myocardial cell to perish weakly effectively, increases along with the density, suppresses the weak perishing rate degree to be stronger.2. the traditional Chinese medicine falls in sugar three Huang Pian the dosage to be possible remarkable under to raise the sugar induction the myocardial cell TGF-β1expression, its mechanism is possible and the upward is the membrane cell's GLUT1expression, subsequently activates protein kinase C (PKC) the amino sugar circuit to concern.
引文
[1]http://tech.qq.com/a/20071120/000288.htm 2007.11.20
[2]Rubler S, et al. New type of cardiomyopathy associated with diabetic glomerulosclerosis. Am J Cardiol,1972,30:595.
[3]RESLM, HULSMANN M, PAACHERR, et al. Heart failure in diabetes. Wien Med Wochenschr[J]. 2009,159(5-6):134-140.
[4]林乐迎,李鸿玮等.糖尿病心肌病心室重塑及心肌细胞凋亡的研究进展.四川解剖学杂志.2010,18(1)：20-21.
[5]Yamagishi SI, Nakamura K, Matsui T, et al. Role of postprandial hyperglycaemia in cardiovascular disease in diabetes. Int J Clin Pract,2007; 61(1):83-87
[6]胡东鹏,倪青.糖尿病心脏病中医诊疗思路与方法[J].中国中医药信息杂志.2006,13(1)：90-91.
[7]胡东鹏,倪青.巧定病性明标本中西合参论治疗——辨治糖尿病心脏病的经验[J].辽宁中医杂志.2007,7(7)：289-290.
[8]杨晓晖,吕仁和.试论络脉病变是早期糖尿病心脏病的病理基础[J].北京中医药大学学报.2005,28(3)：85-86.
[9]杨晓晖,吕仁和.糖尿病心脏病的中医分期辨治探讨[J].北京中医.2006,25(7)：403-405.
[10]杨晓晖.吕仁和教授运用加为四逆散治疗消渴病并发症经验.中医函授通讯.1995,14(4)：32-34
[11]王学良,朱章志,熊曼琪等.从心胃相关论治糖尿病心脏病[J.]新中医.2009,41(2)：5-6.
[12]熊曼琪.加味桃核承气汤(片)治疗糖尿病的临床疗效观察[J].新中医.2009,41(2)
[13]熊曼琪,梁柳文,林安钟等.加为桃核承气汤治疗2型糖尿病的临床与实验研究[J].中西医结合杂志,1997,17(3)：165-168.
[14]张瑞玲,赵丽,马亚丽.心钠素对老年心血管疾病气虚血瘀证诊断的意义[J].中国中医药信息杂志.2006,13(1)：90-91.
[15]消渴病中医辨证分型与诊疗方案[C].第三届中国中医药学会消渴病专业委员会学术交流论文汇编.1992 1
[16]胡继玲.消渴病心病证治规律的临床研究[J].北京中医药大学硕十研究生学位论文.1992
[17]徐莉嘉,凌在荣,王燕.糖尿病合并冠心病辨治体会[J].新中医,2007,39(12)：5-6.
[18]付春梅,郭力,王梅.甘露降糖散治疗糖尿病心脏病的临床观察[J].中医药信息,2004,19(4)：90-91.
[19]陈晓雯.益气活血方治疗糖尿病无症状心肌缺血性心脏病临床观察[J].实用中西医结合杂志.1994：,11(1)：21.
[20]黄建辉等.糖尿病慢性并发症的中医治疗与展望[J].福建中医药1991,22(6)：58.
[21]糖尿病中医防治指南中国中医药出版社.2007第一版
[22]上官海娟,徐江,官洪山等.当归对心肌梗死后心肌细胞凋亡和心室重构的影响[J].中国中西医结合急救杂志2008,15(1)：39-43.
[23]刘洪,许惠琴,时艳.山萸肉环烯醚萜总苷对2型糖尿病心脏病大鼠胰岛素抵抗及血脂含量 的影响[J].中药药理与临床,2007：17(8)：62.
[24]陈蔚,叶红英,俞茂华.黄芪多糖对糖尿病心肌胶原代谢的影响[J].上海交通大学学报,2007：27(10)：1206-1209.
[25]杜立建,吴俊喜.心室重构的中医单味药治疗研究进展[J].现代中西医结合杂志.2007,16(2)：282-283.
[26]葛敏,刘彤,关宿东等.绞股蓝总苷对糖尿病心肌病大鼠心脏功能的影响[J].沈阳药科大学学报,2007,24(6)：355-358.
[27]姜永涛.辽东楤木总苷对大鼠实验性心肌缺血的保护作用[J].中国药理学与毒理学杂志,1998.2(1)：20.
[28]周重楚.龙牙楤木总苷对变态反应的影响[J].中国药理学与毒理学杂志,1991,5(1)：30.
[29]邓汉武.辽东葱木总苷对大鼠实验性心肌缺血的保护作用[J].中国药理学与毒理学杂志,1998.2(1)：20.
[30]奚树刚,张学新,张文杰等.龙芽葱木总皂苷对大鼠早期糖尿病心肌病的保护作用及机制[J].吉林大学学报,2008：34(2)：289-290.
[31]Hamby RI, Zoneraish S, Sherman L, et al. Diabetic cardiomyopathy[J]. TAMA 1974, 229 (13):1749-1754.
[32]Fonarow GC, Srikanthan P. Diabetic Cardiomyopathy. Endocrinol Metab Clin North Am, 2006; 35(3):575-599
[33]Poornima IG, Parikh P, Shannon RP. Diabetic Cardiomyopathy:the search for a unifying hypothesis. Cir Res 2006;98(5):596-605.
[34]Ronghua Yang, Lili A. barouch Leptin Signaling and Obesity Cardiovascular Consequences. Circulation Research.2007;101:545-559.
[35]McGafin KR, et al. Leptin signaling reduces the severity of cardiac dysfunction and remodeling after chronic ischaemic injury. Cardiovascular Research 2008;77(1):54-63.
[36]Yang R, Barouch LA. Leptin signaling and obesity:cardiovascular consequences. Circ Res 2007;101(6):545-59.
[37]Dobrin JS, Lebeche D. Diabetic cardiomyopathy, causes and effects. Rev Endoer Meatb Disord.2010; 11(1):31-9.
[38]Scherer PE. Adipose tissue:from lipid storage compartment to endocrine organ. Diabetes 2006;55(6):1537-45.
[39]Kadowaki T, et al. Adiponectin and adiponectin receptors in insulin resistance, diabetes,and the metabolic syndrome. J Clin Invest 2006;116(7):1784-92.
[40]Wang ZV, Scherer PE. Adiponectin, cardiovascular function, and hypertension. Hypertension 2008:51(1):8-14.
[41]Shibata R, et al. Adiponectin-mediated modulation of hypertrophic signals in the heart. Nat Med 2004;10(12):1384-9.
[42]Shibata R, et al. Adiponectin protects against myocardial ischemia-reperfusion injury through AMPK- and COX-2-dependent mechanisms. Nat Med 2005:11(10):1096-103.
[43]李秀典,李显着.蛋白激酶C激活与糖尿病心脏病[J].中国现代临床医学杂志,2008,5(7)：48-52.
[44]Olivares-Reyes JA, Arellano-Placncarte A, Castillo-Hernandez JR. Angiotensin II and the development of insulin resistance:implications for diabetes. Mol Cell Endocrinol.2009;302:128-39.
[45]Zheng M, Reynolds C, Jo SH, et al, Intracellularacidos-activated p38 MAPK signaling and its essential role cardiomyocyte hypoxic injury [J]. FASEB J,2005,1(1):109-111.
[46]Horowitz JD, Chirkov YY, kennedy JA, SverdlovAL. Modulation of myocardial metabolism: an emerging therapeutic principle. Curr Opin Cardiol.2010:25:329-34.
[47]·Kim AS, Miller EJ, Young LH. AMP-activated protein kinase:a core signaling pathway in the heart. Acta Physiol (Oxf) 2009; 196(1):37-53.
[48]Massague J. The TGF-β family of growth and differentiation factors [J]. Cell, 1987,49:437.
[49]Bottinger E P, Bizer M. TGF-β signaling in renal disease [J]. Jam Soc Nephrol, 2002,1310:2600.
[50]Sharm K, Ziyadeh F N, Alzahabi B, et al. Increased renal production of transforming growth factor-beta 1 in patients with type 2 diabetes [J]. Diabetes,1997,46:854.
[51][Laviades C, Varo N,Diez J. Transforming growth factor beta in hypertensives with cardiorenal damage[J]. Hypertension,2000,36 (4):517-522.]
[52][Zhong H, Fan JM, Li Z, et al. Effects of hepatocyte growth factor on TGF-betal triggered tubular epithelial-myofibroblast transdifierendation by CTGF in vitro [J]. Sichuan DaXue Xue BaoYi Xue Ban,2005,36(5):653-656.]
[53]Parving HH, Jacobser P. Rossing K, et al. Rossing benefits of long-term antihypertensive treatment on prognosis in diabetic Dephropathy. Kidney Int,1996,49:1778-1782.
[54]Gaml6.Gambaro G, Ceol M,Del prete D, et al.GLUT-1 and TGF beta:the link between hyperglycaemia and diabetic nephropathy. Nephrol Dial Transplant,2000,15(9):1476-1477.
[55]Li JP, Petrov V, Rumilla K, et al. Transforming growth factor betal promotes contraction of collagen gel by cardie fibroblasts through their differentiation into myofibroblasts[J]. Methods Find EXP Clin Pharmacol,2003,25(2):79-86.
[56][Paulus WJ. Cytokines and heart failure[J]. Heart Fail Monit,2000,1 (2):50-56]
[57][刘向群,刘传亮.血清TNF-α, STNFRI, IL-10, TGF-β1水平与充血性心力衰竭心功能的相关性研究[J].山东大学学报,2005,43(1)：73-74]
[58]孙静平脾胃与冠心病关系初探[J].中华内科杂志,1983,2212：737-738
[59]胡东鹏,倪青.糖尿病心脏病中医诊疗思路与方法[J].中国中医药信息杂志.2006,13(1)：90-91.
[60]糖尿病中医防治指南中国中医药出版社.2007第一版
[61]王慧瑞.降糖三黄片干预糖尿病心肌病早期心室重构的作用机理研究.博士论文.广州中医药大学,42,2010.
[62]孙静平.脾胃与冠心病关系初探[J].中华内科杂志,1983,22(12)：737-738.
[63]方剑锋,李赛美,林士毅,王保华降糖三黄片对热瘀互结型代谢综合征的影响[J].中医杂 志,2010，51(7)：607-610.
[64]王慧睿,李赛美,王保华等降糖三黄片对糖尿病心肌病大鼠心室重构的电镜观察[J].中国中医基础医学杂志2011,17(7)：749-751.
[65]阮氏心顺,王保华,李赛美三黄降糖片治疗糖尿病心脏病临床观察.仲景医学求真.
[66]陈云,李赛美,王保华等.降糖三黄片对糖尿病大鼠肾脏转化生长因-β1表达的影响[J].广州中医药大学学报.2012,29(2)：154-157.
[67]Hiroko Iwama, Sakae Amagaya and Yukio Ogihara. Effect of shosaikoto a Japanese and Chinese traditional herbal medicinal mixture, on the mitogenic activity of lipopolysaccharide:a new pharm acological testing method [J]. Journal of Ethnopharm acology,1987,21:45-53.
[68]王国佐,葛金文.血清药理学方法在中药研究中的进展[J].湖南中医药大学学报,2007,6(27)：78-80.
[69]薛岚.近年中药药理研究进展[J].中药药理与临床,2000,16(1)：46-48.
[70]崔晓兰,贺玉琢,高英杰.中药药理研究的新思路——中药血清药理学.中国中医药科技,1997，4(4)：239-250.
[71]蒙一纯,丁霞,贲长恩等.中药血清药理学应用研究展望[J].北京中医药大学学报,1999,7(22)：42-43.
[72]米永杰,李健.中药血清药理学研究概述[J].四川解剖学杂志,2006,14(4)：34-35.
[73]刘烨.关于中药血清药理学实验方法的讨论[J].贵阳中医学院学报,2004,26(1)：53-55.
[74]Lu H J. The several approach on study of serum pharmacology[J]. Primary Journal of Chinese Materia Medica.2000,14(3):51-52.
[75]Wang N S, LEI Yan, LIU Ping, et al. Consideration on the serum pharmacology [J]. Chin J Integrated Tradit West Med,1999,19(5):263.
[76]王力倩.用血清药理学方法研究中药苦参、仙鹤草的抗肿瘤作用[J].中国中医药科技,1995,3(5)： 19-21.
[77]李仪奎.中药血清药理学实验方法的若干问题[J].中药新药与临床药理,1999,10(2)：95-98.
[78]王筠,张军平.中药血清药理学实验方法研究概况[J].天津中医药,2005,2(22)：86-88.
[79]李振光,王净净.关于中药血清药理学方法的思考[J].中国中医药信息杂志,2002,9(2)：5.
[80]刘倩,马学盛.化浊行血汤含药血清血管内皮保护作用及其体内给药时效差异的研究[J].时珍国医国药,2009,20(10)：2501.
[81]魏雅芹,汤道权,高媛媛,等.银杏叶提取物血清药理学方法的建立及验证[J].中药药理与临床,2009,25(3)：74.
[82]李秀荣,张丹,齐元富,等.消瘤平移合剂含药血清诱导肝癌H-7402细胞凋亡的实验研究[J].中国中西医结合杂志,2001,21(9)：684.
[83]黄臣虎,陆茵,高骁君等.中药血清药理学研究进展[J].中国方剂实验学杂志,2011,5(10)：266-271.
[84]殷飞,姚树坤,吴新满,等.清肝化瘀方药含药血清对肝癌细胞SMMC-7721增殖和凋亡的影响[J].中药药理与临床.
[85]罗琳,窦志华,丁安伟,等.大鼠灌胃复方五仁醇胶囊后血清和血浆中五味子醇甲和五味子 乙素的比较[J].中草药,2006,37(10)：1486.
[86]Matoba S, Tat sum i T, Keira N, et al. Cardio protective effect of ang iotensin-converting enzyme inhibition against hypoxia/reoxygenation injury incultred rat cardiac myocytes[J]. Circulation,1999,99 (6):817-822.
[87]王晓明,臧益民,高峰等Staurosporine诱导小鼠心肌细胞凋亡模型的构建[J].心脏杂志,2004,16(1)：4-10.
[88]梁家亮,余细勇,单志新等.高糖诱导H9c2心肌细胞凋亡并激活JNK通路的研究[J].实用医学杂志,2010,26(12)：2094-2097.
[89]潘春留,闫雷,娜仁图娜拉等.流式细胞术研究细胞凋亡的方法与技术[J].现代生物医学进展.2008，9(4)：790-793·
[90]Bertoluci MC, Schmid H, Lachat JJ, et al. Transforming growth factor-beta in the development of rat diabetic nephropathy. A 10-month study with insulin-treated rats[J].Nephron,1996,74(1):189-196
[91]World Health Organization. Definition,diagnosis and classification of diabetes mellitus and its complication, report of a WHO consultation[R]. Geneva, World Health Organization,1999,1.
[92]第一届全国内科学术会议心血管病组.关于冠状动脉性心脏病命名及诊断标准的建议[J].中华心血管病杂志,1981,9(1)：75-76.
[93]李易崇.糖尿病心脏病临床证治规律研究.硕士论文.广州中医药大学,37,2006.
[94]李赛美.糖尿病湿热证及糖尿病心脏病证治规律理论与临床研究.博士论文.广州中医药大学.58,2005
[2]Rubler S, et al. New type of cardiomyopathy associated with diabetic glomerulosclerosis. Am J Cardiol,1972,30:595.
[3]RESLM, HULSMANN M, PAACHERR, et al. Heart failure in diabetes. Wien Med Wochenschr[J]. 2009,159(5-6):134-140.
[4]林乐迎,李鸿玮等.糖尿病心肌病心室重塑及心肌细胞凋亡的研究进展.四川解剖学杂志.2010,18(1)：20-21.
[5]Yamagishi SI, Nakamura K, Matsui T, et al. Role of postprandial hyperglycaemia in cardiovascular disease in diabetes. Int J Clin Pract,2007; 61(1):83-87
[6]胡东鹏,倪青.糖尿病心脏病中医诊疗思路与方法[J].中国中医药信息杂志.2006,13(1)：90-91.
[7]胡东鹏,倪青.巧定病性明标本中西合参论治疗——辨治糖尿病心脏病的经验[J].辽宁中医杂志.2007,7(7)：289-290.
[8]杨晓晖,吕仁和.试论络脉病变是早期糖尿病心脏病的病理基础[J].北京中医药大学学报.2005,28(3)：85-86.
[9]杨晓晖,吕仁和.糖尿病心脏病的中医分期辨治探讨[J].北京中医.2006,25(7)：403-405.
[10]杨晓晖.吕仁和教授运用加为四逆散治疗消渴病并发症经验.中医函授通讯.1995,14(4)：32-34
[11]王学良,朱章志,熊曼琪等.从心胃相关论治糖尿病心脏病[J.]新中医.2009,41(2)：5-6.
[12]熊曼琪.加味桃核承气汤(片)治疗糖尿病的临床疗效观察[J].新中医.2009,41(2)
[13]熊曼琪,梁柳文,林安钟等.加为桃核承气汤治疗2型糖尿病的临床与实验研究[J].中西医结合杂志,1997,17(3)：165-168.
[14]张瑞玲,赵丽,马亚丽.心钠素对老年心血管疾病气虚血瘀证诊断的意义[J].中国中医药信息杂志.2006,13(1)：90-91.
[15]消渴病中医辨证分型与诊疗方案[C].第三届中国中医药学会消渴病专业委员会学术交流论文汇编.1992 1
[16]胡继玲.消渴病心病证治规律的临床研究[J].北京中医药大学硕十研究生学位论文.1992
[17]徐莉嘉,凌在荣,王燕.糖尿病合并冠心病辨治体会[J].新中医,2007,39(12)：5-6.
[18]付春梅,郭力,王梅.甘露降糖散治疗糖尿病心脏病的临床观察[J].中医药信息,2004,19(4)：90-91.
[19]陈晓雯.益气活血方治疗糖尿病无症状心肌缺血性心脏病临床观察[J].实用中西医结合杂志.1994：,11(1)：21.
[20]黄建辉等.糖尿病慢性并发症的中医治疗与展望[J].福建中医药1991,22(6)：58.
[21]糖尿病中医防治指南中国中医药出版社.2007第一版
[22]上官海娟,徐江,官洪山等.当归对心肌梗死后心肌细胞凋亡和心室重构的影响[J].中国中西医结合急救杂志2008,15(1)：39-43.
[23]刘洪,许惠琴,时艳.山萸肉环烯醚萜总苷对2型糖尿病心脏病大鼠胰岛素抵抗及血脂含量 的影响[J].中药药理与临床,2007：17(8)：62.
[24]陈蔚,叶红英,俞茂华.黄芪多糖对糖尿病心肌胶原代谢的影响[J].上海交通大学学报,2007：27(10)：1206-1209.
[25]杜立建,吴俊喜.心室重构的中医单味药治疗研究进展[J].现代中西医结合杂志.2007,16(2)：282-283.
[26]葛敏,刘彤,关宿东等.绞股蓝总苷对糖尿病心肌病大鼠心脏功能的影响[J].沈阳药科大学学报,2007,24(6)：355-358.
[27]姜永涛.辽东楤木总苷对大鼠实验性心肌缺血的保护作用[J].中国药理学与毒理学杂志,1998.2(1)：20.
[28]周重楚.龙牙楤木总苷对变态反应的影响[J].中国药理学与毒理学杂志,1991,5(1)：30.
[29]邓汉武.辽东葱木总苷对大鼠实验性心肌缺血的保护作用[J].中国药理学与毒理学杂志,1998.2(1)：20.
[30]奚树刚,张学新,张文杰等.龙芽葱木总皂苷对大鼠早期糖尿病心肌病的保护作用及机制[J].吉林大学学报,2008：34(2)：289-290.
[31]Hamby RI, Zoneraish S, Sherman L, et al. Diabetic cardiomyopathy[J]. TAMA 1974, 229 (13):1749-1754.
[32]Fonarow GC, Srikanthan P. Diabetic Cardiomyopathy. Endocrinol Metab Clin North Am, 2006; 35(3):575-599
[33]Poornima IG, Parikh P, Shannon RP. Diabetic Cardiomyopathy:the search for a unifying hypothesis. Cir Res 2006;98(5):596-605.
[34]Ronghua Yang, Lili A. barouch Leptin Signaling and Obesity Cardiovascular Consequences. Circulation Research.2007;101:545-559.
[35]McGafin KR, et al. Leptin signaling reduces the severity of cardiac dysfunction and remodeling after chronic ischaemic injury. Cardiovascular Research 2008;77(1):54-63.
[36]Yang R, Barouch LA. Leptin signaling and obesity:cardiovascular consequences. Circ Res 2007;101(6):545-59.
[37]Dobrin JS, Lebeche D. Diabetic cardiomyopathy, causes and effects. Rev Endoer Meatb Disord.2010; 11(1):31-9.
[38]Scherer PE. Adipose tissue:from lipid storage compartment to endocrine organ. Diabetes 2006;55(6):1537-45.
[39]Kadowaki T, et al. Adiponectin and adiponectin receptors in insulin resistance, diabetes,and the metabolic syndrome. J Clin Invest 2006;116(7):1784-92.
[40]Wang ZV, Scherer PE. Adiponectin, cardiovascular function, and hypertension. Hypertension 2008:51(1):8-14.
[41]Shibata R, et al. Adiponectin-mediated modulation of hypertrophic signals in the heart. Nat Med 2004;10(12):1384-9.
[42]Shibata R, et al. Adiponectin protects against myocardial ischemia-reperfusion injury through AMPK- and COX-2-dependent mechanisms. Nat Med 2005:11(10):1096-103.
[43]李秀典,李显着.蛋白激酶C激活与糖尿病心脏病[J].中国现代临床医学杂志,2008,5(7)：48-52.
[44]Olivares-Reyes JA, Arellano-Placncarte A, Castillo-Hernandez JR. Angiotensin II and the development of insulin resistance:implications for diabetes. Mol Cell Endocrinol.2009;302:128-39.
[45]Zheng M, Reynolds C, Jo SH, et al, Intracellularacidos-activated p38 MAPK signaling and its essential role cardiomyocyte hypoxic injury [J]. FASEB J,2005,1(1):109-111.
[46]Horowitz JD, Chirkov YY, kennedy JA, SverdlovAL. Modulation of myocardial metabolism: an emerging therapeutic principle. Curr Opin Cardiol.2010:25:329-34.
[47]·Kim AS, Miller EJ, Young LH. AMP-activated protein kinase:a core signaling pathway in the heart. Acta Physiol (Oxf) 2009; 196(1):37-53.
[48]Massague J. The TGF-β family of growth and differentiation factors [J]. Cell, 1987,49:437.
[49]Bottinger E P, Bizer M. TGF-β signaling in renal disease [J]. Jam Soc Nephrol, 2002,1310:2600.
[50]Sharm K, Ziyadeh F N, Alzahabi B, et al. Increased renal production of transforming growth factor-beta 1 in patients with type 2 diabetes [J]. Diabetes,1997,46:854.
[51][Laviades C, Varo N,Diez J. Transforming growth factor beta in hypertensives with cardiorenal damage[J]. Hypertension,2000,36 (4):517-522.]
[52][Zhong H, Fan JM, Li Z, et al. Effects of hepatocyte growth factor on TGF-betal triggered tubular epithelial-myofibroblast transdifierendation by CTGF in vitro [J]. Sichuan DaXue Xue BaoYi Xue Ban,2005,36(5):653-656.]
[53]Parving HH, Jacobser P. Rossing K, et al. Rossing benefits of long-term antihypertensive treatment on prognosis in diabetic Dephropathy. Kidney Int,1996,49:1778-1782.
[54]Gaml6.Gambaro G, Ceol M,Del prete D, et al.GLUT-1 and TGF beta:the link between hyperglycaemia and diabetic nephropathy. Nephrol Dial Transplant,2000,15(9):1476-1477.
[55]Li JP, Petrov V, Rumilla K, et al. Transforming growth factor betal promotes contraction of collagen gel by cardie fibroblasts through their differentiation into myofibroblasts[J]. Methods Find EXP Clin Pharmacol,2003,25(2):79-86.
[56][Paulus WJ. Cytokines and heart failure[J]. Heart Fail Monit,2000,1 (2):50-56]
[57][刘向群,刘传亮.血清TNF-α, STNFRI, IL-10, TGF-β1水平与充血性心力衰竭心功能的相关性研究[J].山东大学学报,2005,43(1)：73-74]
[58]孙静平脾胃与冠心病关系初探[J].中华内科杂志,1983,2212：737-738
[59]胡东鹏,倪青.糖尿病心脏病中医诊疗思路与方法[J].中国中医药信息杂志.2006,13(1)：90-91.
[60]糖尿病中医防治指南中国中医药出版社.2007第一版
[61]王慧瑞.降糖三黄片干预糖尿病心肌病早期心室重构的作用机理研究.博士论文.广州中医药大学,42,2010.
[62]孙静平.脾胃与冠心病关系初探[J].中华内科杂志,1983,22(12)：737-738.
[63]方剑锋,李赛美,林士毅,王保华降糖三黄片对热瘀互结型代谢综合征的影响[J].中医杂 志,2010，51(7)：607-610.
[64]王慧睿,李赛美,王保华等降糖三黄片对糖尿病心肌病大鼠心室重构的电镜观察[J].中国中医基础医学杂志2011,17(7)：749-751.
[65]阮氏心顺,王保华,李赛美三黄降糖片治疗糖尿病心脏病临床观察.仲景医学求真.
[66]陈云,李赛美,王保华等.降糖三黄片对糖尿病大鼠肾脏转化生长因-β1表达的影响[J].广州中医药大学学报.2012,29(2)：154-157.
[67]Hiroko Iwama, Sakae Amagaya and Yukio Ogihara. Effect of shosaikoto a Japanese and Chinese traditional herbal medicinal mixture, on the mitogenic activity of lipopolysaccharide:a new pharm acological testing method [J]. Journal of Ethnopharm acology,1987,21:45-53.
[68]王国佐,葛金文.血清药理学方法在中药研究中的进展[J].湖南中医药大学学报,2007,6(27)：78-80.
[69]薛岚.近年中药药理研究进展[J].中药药理与临床,2000,16(1)：46-48.
[70]崔晓兰,贺玉琢,高英杰.中药药理研究的新思路——中药血清药理学.中国中医药科技,1997，4(4)：239-250.
[71]蒙一纯,丁霞,贲长恩等.中药血清药理学应用研究展望[J].北京中医药大学学报,1999,7(22)：42-43.
[72]米永杰,李健.中药血清药理学研究概述[J].四川解剖学杂志,2006,14(4)：34-35.
[73]刘烨.关于中药血清药理学实验方法的讨论[J].贵阳中医学院学报,2004,26(1)：53-55.
[74]Lu H J. The several approach on study of serum pharmacology[J]. Primary Journal of Chinese Materia Medica.2000,14(3):51-52.
[75]Wang N S, LEI Yan, LIU Ping, et al. Consideration on the serum pharmacology [J]. Chin J Integrated Tradit West Med,1999,19(5):263.
[76]王力倩.用血清药理学方法研究中药苦参、仙鹤草的抗肿瘤作用[J].中国中医药科技,1995,3(5)： 19-21.
[77]李仪奎.中药血清药理学实验方法的若干问题[J].中药新药与临床药理,1999,10(2)：95-98.
[78]王筠,张军平.中药血清药理学实验方法研究概况[J].天津中医药,2005,2(22)：86-88.
[79]李振光,王净净.关于中药血清药理学方法的思考[J].中国中医药信息杂志,2002,9(2)：5.
[80]刘倩,马学盛.化浊行血汤含药血清血管内皮保护作用及其体内给药时效差异的研究[J].时珍国医国药,2009,20(10)：2501.
[81]魏雅芹,汤道权,高媛媛,等.银杏叶提取物血清药理学方法的建立及验证[J].中药药理与临床,2009,25(3)：74.
[82]李秀荣,张丹,齐元富,等.消瘤平移合剂含药血清诱导肝癌H-7402细胞凋亡的实验研究[J].中国中西医结合杂志,2001,21(9)：684.
[83]黄臣虎,陆茵,高骁君等.中药血清药理学研究进展[J].中国方剂实验学杂志,2011,5(10)：266-271.
[84]殷飞,姚树坤,吴新满,等.清肝化瘀方药含药血清对肝癌细胞SMMC-7721增殖和凋亡的影响[J].中药药理与临床.
[85]罗琳,窦志华,丁安伟,等.大鼠灌胃复方五仁醇胶囊后血清和血浆中五味子醇甲和五味子 乙素的比较[J].中草药,2006,37(10)：1486.
[86]Matoba S, Tat sum i T, Keira N, et al. Cardio protective effect of ang iotensin-converting enzyme inhibition against hypoxia/reoxygenation injury incultred rat cardiac myocytes[J]. Circulation,1999,99 (6):817-822.
[87]王晓明,臧益民,高峰等Staurosporine诱导小鼠心肌细胞凋亡模型的构建[J].心脏杂志,2004,16(1)：4-10.
[88]梁家亮,余细勇,单志新等.高糖诱导H9c2心肌细胞凋亡并激活JNK通路的研究[J].实用医学杂志,2010,26(12)：2094-2097.
[89]潘春留,闫雷,娜仁图娜拉等.流式细胞术研究细胞凋亡的方法与技术[J].现代生物医学进展.2008，9(4)：790-793·
[90]Bertoluci MC, Schmid H, Lachat JJ, et al. Transforming growth factor-beta in the development of rat diabetic nephropathy. A 10-month study with insulin-treated rats[J].Nephron,1996,74(1):189-196
[91]World Health Organization. Definition,diagnosis and classification of diabetes mellitus and its complication, report of a WHO consultation[R]. Geneva, World Health Organization,1999,1.
[92]第一届全国内科学术会议心血管病组.关于冠状动脉性心脏病命名及诊断标准的建议[J].中华心血管病杂志,1981,9(1)：75-76.
[93]李易崇.糖尿病心脏病临床证治规律研究.硕士论文.广州中医药大学,37,2006.
[94]李赛美.糖尿病湿热证及糖尿病心脏病证治规律理论与临床研究.博士论文.广州中医药大学.58,2005