心虚证大鼠心肌荧光组织化学及超微结构的实验研究

目的:观察心虚证对心肌细胞微细结构及超微结构的影响。方法:选用健康大鼠随机分为造模组和对照组,于造模48h、72h分别取心肌组织作冰冻切片、超薄切片,采用荧光组织化学及电镜技术,观察心肌细胞内糖原和DNA的分布、含量及超微结构的变化。结果:心虚证造模组大鼠的心肌细胞在48h后过碘酸-Schiff反应(荧光PAS反应)明显减弱,72h后DNA荧光反应阳性物明显低于对照组;电镜观察心肌细胞内线粒体肿胀、变形,嵴断裂,数量减少.基质密度降低,肌原纤维有断裂,糖原减少。经立体学定量分析,线粒体体密度与对照组相比差异有显著性(P<0.01)。结论:心虚证可导致心肌细胞的损伤,影响心肌代谢及细胞结构。本研究为心虚证的病因病机学提供一定的实验依据,可指导临床治疗。

大鼠心脏腺苷预处理心肌组化和细胞化学变化

目的：应用图象分析和电镜细胞化学方法，观察大鼠心脏腺苷预处理心肌琥珀酸脱氢酶、细胞色素氧化酶及Ｃａ２＋Ｍｇ２＋－ＡＴＰａｓｅ活性变化。方法：采用ＳＤ雄性大鼠３２只分４组，即正常对照组（ＮＣ），缺血／再灌组（Ｉ／Ｒ），缺血预处理组，腺苷预处理组。用图象分析检测琥珀酸脱氢酶、细胞色素氧化酶和Ｃａ２＋Ｍｇ２＋－ＡＴＰａｓｅ 活性的变化；用电镜细胞化学方法检测Ｃａ２＋Ｍｇ２＋－ＡＴＰａｓｅ活性。结果：在缺血预处理组及腺苷预处理组琥珀酸脱氢酶、细胞色素氧化酶、Ｃａ２＋Ｍｇ２＋－ＡＴＰａｓｅ活性均高于 Ｉ／Ｒ组。结论：大鼠心脏腺苷预处理能提高心肌琥珀酸脱氢酶、细胞色素氧化酶Ｃａ２＋Ｍｇ２＋－ＡＴＰａｓｅ活性。

碱性成纤维细胞生长因子基因对心肌梗死后冠状血管新生及血管结构的影响

为探讨心肌内注射碱性成纤维细胞生长因子基因对心肌梗死后冠状血管新生和血管结构的影响。以碱裂解法大量制备质粒 ;采用开胸结扎兔冠状动脉左前室间支法 ,建立兔急性前壁心肌梗死模型。模型制备成功后将动物分为治疗组 (n =1 9)和对照组 (n =1 8) ,并于心肌内分别注射pcDNA3 碱性成纤维细胞生长因子 1 0 0 μg和pcDNA31 0 0 μg,饲养至第 2、6和 1 2周末处死 ;免疫组织化学法观察蛋白表达 ;病理切片观察梗死心肌组织学变化、缺血心肌内血管新生和血管管壁及管腔变化情况。结果发现 :(1 )实验中描记的心电图证实兔急性心肌梗死模型制作成功。 (2 )免疫组织化学观察注射pcDNA3 碱性成纤维细胞生长因子处心肌组织在 6周内有碱性成纤维细胞生长因子蛋白的表达。 (3)病理切片行图象分析计算血管密度发现 :治疗组毛细血管密度和小动脉密度显著高于对照组。 (4 )实验第 6周治疗组与对照组动脉平均管壁厚度分别为 1 9.8± 9.9μm比 1 8.9± 9.6 μm (P >0 .0 5 ) ;1 2周分别为 2 8.3± 1 1 .5 μm比 2 4 .1± 1 1 .3μm (P <0 .0 1 ) ;6周比值分别为 0 .31± 0 .1 6比 0 .2 4± 0 .1 2 (P <0 .0 1 ) ;1 2周分别为 0 .34± 0 .1 5比 0 .2 5± 0 .0 9(P <0 .0 1 )。实验结果提示 ,心肌内注射碱性成纤维细胞生长?

Ultrastructure and histochemistry of rat myocardial capillary endothelial cells in response to diabetes and hypertension

Insufficient growth and rarefaction of capillaries, followed by endothelial dysfunction may represent one of the most critical mechanisms involved in heart damage. In this study we examined histochemical and ultrastructural changes in myocardial capillary endothelium in two models of heart failure streptozotocin-induced diabetes mellitus （STZ） and NO-deficient hypertension in male Wistar rats. Diabetes was induced by a single i.v. dose of STZ （45 mg/kg） and chronic 9-week stage was analysed. To induce NO-deficient hypertension, animals were treated with inhibitor of NO synthase Lnitroarginine methylester （L-NAME） （40 mg/kg） for 4 weeks. Left ventricular tissue was processed for enzyme catalytic histochemistry of capillary alkaline phosphatase （A1Ph）, dipeptidyl peptidase IV （DPP IV）, and endothelial NO synthase/NADPH-diaphorase （NOS） and for ultrastructural analysis. In diabetic and hypertensive rats, lower/absent A1Ph and DPP IV activities were found in focal micro-areas. NOS activity was significantly reduced and persisted only locally. Quantitative evaluation demonstrated reduction of reaction product intensity of A1Ph, DPP and NOS by 49.50%,74.36%, 20.05% in diabetic and 62.93%, 82.71%, 37.65% in hypertensive rats. Subcellular alterations of endothelial cells were found in heart of both groups suggesting injury of capillary function as well as compensatory processes. Endothelial injury was more significant in diabetic animals, in contrast the adaptation was more evident in hypertensive ones. Concluding： both STZ-induced diabetes- and NO-deficient hypertension-related cardiomyopathy were accompanied by similar features of structural remodelling of cardiac capillary network manifested as angiogenesis and angiopathy. The latter was however, predominant and may accelerate disappearance of capillary endothelium contributing to myocardial dysfunction.

Predictors of long-term outcome in patients with biopsy proven inflammatory cardiomyopathy

Background The objective of this study was to identify prognostic indicators in patients with inflammatory cardiomyopathy （iCM） on endomyocardial biopsy （EMB）. Methods and results Between 2007 and 2011 all consecutive patients with diagnosed with iCM at EMB were retrospectively analyzed. The combined primary endpoint （EP） （1E°P） was cardiac death, aborted sudden cardiac death/appropriate implantable cardioverter defibrillator （ICD） shock, progressive heart failure requiring left venWicular assist device （LVAD） implantation and heart transplantation. 503 patients （mean age 58 ° 12 years, 73% male） were available for analysis. Genomes of cardiotrophic viruses were detected in 396 patients （79%） and immuno-histochemical signs of inflammation were present in 223 individuals （44%）. After 3.6 ° 2.4 years of follow-up, cardiac mortality was 3.0% （n = 14） and a total of 8.6% （n = 40） reached the primary endpoint. Independent predictors for the 1 °EP were： age 〉 50 years, presence and duration （〈 28 days） of symptomatic heart failure. A risk stratification approach based on the results of the multivariate analysis demonstrated that absence of signs and/or symptoms of congestive heart failure in younger （〈 50 years） patients with longer （〉 28 days） duration of disease appear to have an excellent prognosis with 100% survival and no events during follow-up The presence of all above mentioned independent risk factors results in an 1°EP occurrence of 35.9%. Conclusions Symptoms of heart fail- ure, short duration of disease, and older age are indicators of poor outcome in patients with iCM.

Therapeutic Effect of Zygophyllum cornutum on Metabolic Disturbances, Oxidative Stress in Heart Tissue and Histological Changes in Myocardium of Streptozotocin-induced Diabetic Rats

The present study depicts the therapeutic effect of Zygophyllum cornutum methanolic extract （ZCME） on metabolic disturbances, oxidative stress in heart and myocardium histological changes in streptozotocin （STZ）-induced diabetic rats. Three days after diabetes induction, ZCME was administered orally for six weeks （700 mg/kg bw/day）. Serum glucose and lipid profile were evaluated. Reduced glutathione （GSH）, catalase （CAT） and thiobarbituric acid reactive substances （TBARS） were measured in heart tissue. The results showed increased levels of blood glucose, total cholesterol （TC）, LDL cholesterol （LDL-C） and triglyceride （TG） in the diabetic rats. On the other hand, the level of HDL cholesterol （HDL-C） decreased. Compared to the control normal rats, the level of TBARS in heart tissue was markedly increased while GSH and CAT were significantly modified in the diabetic rats. Oral administration of ZCME has normalized serum glucose and lipid profile. TBARS were significantly reduced in heart while CAT and GSH were markedly normalized. Myocardium sections showed the absence of histological changes observed in the diabetic rats. The study suggests that Zygophyllum cornutum may provide a useful therapeutic option in the reversal of metabolic disturbances and oxidative stress-induced cardiac dysfunction in diabetes mellitus.

Effects of Panax Notoginseng Saponins on Homing of C-kit^+ Bone Mesenchymal Stem Cells to the Infarction Heart in Rats

Objective:To investigate the effects of panax notoginseng saponins(PNS) on homing of C-kit+ bone mesenchymal stem cells(BMSCs) to the infarction heart.Methods:The acute myocardial infraction(AMI) model was established in 140 Wistar rats,105 model rats survived after operation,and the model rats were randomly divided into five groups,21 rats in each group:Western medicine group mobilized by subcutaneous injection of human granuloctye colony stimulating factor(G-CSF) 50 μg·kg-1·d-1;sham operation group and a model group treated by subcutaneous injection of normal saline 50 μg·kg-1·d-1;Chinese medicine group mobilized by intraperitoneal injection of Xuesaitong(血塞通)(ingredients of PNS) 150 mg·kg-1·d-1;integrative medicine group mobilized by subcutaneous injection of G-CSF 50 μg·kg-1·d-1 and intraperitoneal injection of Xuesaitong 150 mg·kg-1·d-1.Except for the sham-operated group,each group was divided into three sub-groups by three time points of 1 d,7 d and 14 d.G-CSF was injected once a day for 7 d.Xuesaitong was injected once a day until the rats were killed.The flow cytometry was used for detection of C-kit + cells in the peripheral blood in different time points,and immunohistochemical method was used for detection of the changes of C-kit + cell and Ki-67+ cell numbers in the marginal zone of AMI.Results:Twenty-four hours after the operation,C-kit + cells had a slight increase in the model group compared with the sham operation group(P>0.05).The peripheral blood C-kit+ cells in the integrative group increased significantly compared with the other groups on 7 d and 14 d(all P<0.05).Meanwhile the expression of C-kit + cells and Ki-67+ cells in the marginal zone of AMI in the integrative group increased significantly compared with the Chinese medicine group,the western medicine group and the model group on 1 d,7 d and 14 d(all P<0.05),and the cells in the integrative group decreased significantly on 14 d compared with that on 7 d(P<0.05).Conclusion:PNS can cooperate with G-CSF to mobilize C-kit+ BMSCs from the marrow into the peripheral blood and promote them "homing" to the infarction heart.