顿抑心肌组织降钙素基因相关肽的免疫组织化学研究

采用大鼠心肌顿抑模型,用免疫组织化学方法观察了顿抑心肌组织中降钙素基因相关肽（ＣＧＲＰ）的改变,结果发现心脏中存在ＣＧＲＰ阳性免疫神经纤维,尤其是心房,冠脉周围,乳头肌以及心室肌中,顿抑心肌中ＣＧＲＰ纤维密度显著减少。     

降钙素基因相关肽对顿抑心肌心功能的影响

建立大鼠心肌顿抑模型,观察降钙素基因相关肽（ＣＧＲＰ）对顿抑心肌心功能的影响。发现再灌注前给予ＣＧＲＰ,可显著减轻心肌顿抑的程度,加快缺血后心功能障碍的恢复。说明ＣＧＲＰ可部分预防心肌顿抑的发生     

降钙素基因相关肽对顿抑心肌心功能的影响

建立大鼠心肌顿抑模型,观察降钙素基因相关肽（ＣＧＲＰ）对顿抑心肌心功能的影响。发现再灌注前给予ＣＧＰＲ,可显著减轻心肌顿抑的程度,加快缺血后心功能障碍的恢复。说明ＣＧＲＰ可部分预防心肌顿抑的发生。     

正常及顿抑心肌组织CGRPmRNA原位杂交研究

在大鼠心肌顿抑模型上采用原位杂交技术观察了正常及顿抑心肌组织降钙素基因相关肽ｍＲＮＡ表达的情况。结果发现正常心房及心室肌组织中存在ＣＧＲＰｍＲＮＡ,心脏的血管周围也有分布。顿抑心肌组织中ＣＧＲＰｍＲＮＡ阳性表达显著减弱。     

正常及顿抑心肌组织CGRPmRNA原位杂交研究

在大鼠心肌顿抑模型上采用原位杂交技术观察了正常及顿抑心肌组织降钙素基因相关肽（ Ｃ Ｇ Ｒ Ｐ）ｍ Ｒ Ｎ Ａ 表达的情况。结果发现正常心房及心室肌组织中存在 Ｃ Ｇ Ｒ Ｐｍ Ｒ Ｎ Ａ,心脏的血管周围也有分布。顿抑心肌组织中 Ｃ Ｇ Ｒ Ｐ ｍ Ｒ Ｎ Ａ 阳性表达显著减弱。     

内皮素和降钙素基因相关肽对异丙肾上腺素致心肌损伤的影响

本实验在大鼠异丙肾上腺素（ＩＳＯ）心肌损伤模型上，观察了内皮素（ＥＴ）及降钙素基因相关肽（ＣＧＲＰ）对ＩＳＯ损伤心肌作用的影响．结果表明：ＥＴ明显加重ＩＳＯ对心肌的损伤，ＣＧＲＰ则能减轻ＩＳＯ对心肌的损伤及ＩＳＯ与ＥＴ对心肌的协同损伤．提示心肌病理情况下，ＥＴ是一种促损伤因素，而ＣＧＲＰ是抗损伤因素．     

大鼠肠系膜动脉床和脊髓降钙素基因相关肽的老年性改变

采用２月龄、１４月龄和１８月龄三个不同年龄阶段的大鼠，在离体灌流的阻力血管组织—肠系膜动脉床和离体温育的脊髓切片上，观察了降钙素基因相关肽（ＣＧＲＰ）在感觉神经外周端和中枢端释放的老年性改变。发现１８月龄的大鼠基础释放及辣椒素刺激后的释放均较２月龄大鼠显著为低；又用免疫组织化学方法对比观察了不同年龄大鼠血管组织中ＣＧＲＰ的分布，发现１４月龄大鼠阻力血管组织中ＣＧＲＰ能神经分布减少；进一步用半定量ＲＴ－ＰＣＲ方法观察到老年大鼠ＣＧＲＰ在合成部位—脊髓背根神经节的ｍＲＮＡ水平增高。证实ＣＧＲＰ组织含量和释放的老年性降低并不是发生于ＣＧＲＰ转录环节。我们推测ＣＧＲＰ的这种改变与老年时血管张力调节失衡，血管阻力增高有关，可能是老年时高血压等心血管疾患发病率增高的机制之一。     

胃泌素释放肽前体对小细胞肺癌诊断作用的初探

胃泌素释放肽 (ＧＲＰ)是脑肠激素的一种。肺癌与ＧＲＰ的关系是 1978年在人胚胎肺的内分泌细胞中发现蛙皮素(ｂｏｍｂｅｓｉｎ)样免疫活性之后开始认识的[1] 。其后的研究表明 ,ＧＲＰ是小细胞肺癌 (ＳＣＬＣ)的重要产物 ,血ＧＲＰ水平是ＳＣＬＣ重要的标志物[2 ] ,但其测定方法繁琐 ,难以普及。研究表明 ,测定胃泌素释放肽前体 (ＰｒｏＧＲＰ)作为ＳＣＬＣ标志物是完全可行的[3 ,4 ] 。ＰｒｏＧＲＰ因部分氨基酸残基的不同 ,存在 3种分子型。利用针对 3种分子的共同序列 ,即ＰｒｏＧＲＰ(31 98)的酶免疫分析药盒 ,测定了 16 2例血清标本 ,…     

降钙素基因相关肽拮抗垂体后叶素引起心肌缺血性电生理改变的观察

用浮置微电极技术观察垂体后叶素（Ｐｉｔ）对兔心肌细胞的异常电生理学效应及降钙素基因相关肽（ＣＧＲＰ）的保护作用。结果表明：静脉注射Ｐｉｔ后，心率减慢，心外膜电图ＳＴ段抬高，Ｔ波倒置，心室肌细胞跨膜动作电位（ＴＭＰ）的静息电位（ＲＰ）及动作电位振幅（ＡＰＡ）明显降低（Ｐ均＜０．０１）。复极化至３０％、５０％和９０％的动作电位时程（ＡＰＤ３０、ＡＰＤ５０、ＡＰＤ９０）亦明显缩短（Ｐ均＜０．０５）。静脉注射Ｐｉｔ同时注射ＣＧＲＰ，可见ＣＧＲＰ明显拮抗以上异常改变，从而起到保护心肌和稳定心肌的电生理作用。     

降钙素基因相关肽对心肌缺血时心脏功能的影响

冠脉内注射降钙素基因相关肽（ＣＧＲＰ）０．３μｇ／ｋｇ，对正常及不同程度冠脉狭窄犬的心功能产生不同影响。结果表明，正常犬冠脉内注射ＣＧＲＰ后，平均动脉压（ＭＡＰ）下降１．２ｋＰａ（Ｐ＜０．０５），同时，心率（ＨＲ）、心输出量（ＣＯ）、左室收缩压峰值（ＬＶＳＰ）均不同程度增加；左室舒张末压（ＬＶＥＤＰ）轻度降低。在中度狭窄３０ｍｉｎ后冠脉内注射ＣＧＲＰ对ＨＲ、ＭＡＰ无明显影响；而重度狭窄后注射ＣＧＲＰ，ＭＡＰ由狭窄时降低逐渐增高，ＨＲ由增快而变慢。ＣＯ、ＬＶＳＰ均显著增高，ＬＶＥＤＰ降低，此作用较冠脉狭窄前更为明显。提示ＣＧＲＰ能扩张冠状动脉，增加冠脉血流量和心排血量，增强心肌收缩力，对缺血心脏功能有保护作用     

Collagen loss in the stunned myocardium.

BACKGROUND. This study was performed to biochemically assess and quantify the previously observed ultrastructural alterations in the collagen matrix of stunned myocardium. METHODS AND RESULTS. The stunned myocardium was produced in 13 mongrel dogs by a series of 12 coronary artery occlusions of 5 minutes followed by 10-minute reperfusion periods, with a final reperfusion period of 90 minutes. Regional systolic function in the stunned myocardium was 17% of control. Relative end-diastolic length in the stunned region increased up to 8%. There was a nonuniform transmural loss of collagen. Hydroxyproline in the stunned endocardium was not different from control. The stunned midwall and epicardium demonstrated 12.5% (p less than 0.05) and 14.6% (p less than 0.005) decreases, respectively. All transmural layers in the stunned myocardium had significant increases in collagenase activity before procollagenase activation, averaging a 73.6% increase (p less than 0.025). Complete activation of all procollagenase forms with aminophenylmercuric acetate revealed no differences in fully activated collagenase between the stunned and normal regions. The lysosomal enzymes, elastase and cathepsin G, were not different between stunned and normal zone tissue. These results would tend to exclude exogenous sources of protease in the stunned myocardium at the 90-minute final reperfusion time frame. Collagen fibers were isolated from the stunned and normal zone tissue and underwent dansyl chloride reaction. Stunned collagen fibers had 9% greater dansyl labeling, suggesting greater numbers of exposed N-terminal amino acid residues on the fiber and compatible with greater enzymatic cleavage activity on the stunned collagen matrix. Tissue water content was consistently greater in the stunned region compared to the normal: a uniform transmural increase of approximately 1.7%. CONCLUSIONS. The stunned myocardium is characterized by both systolic dysfunction and diastolic expansion or dilatation. Endogenous procollagenase is activated by the ischemic process leading to degradation of the extracellular matrix. The underlying mechanisms may be relevant in ischemic enlargement of the heart and cardiomyopathy.     

Innervation of an Esophageal Ectatic Submucosal Blood Vessel in Achalasia and a Comparison with Normals

Achalasia is a disease of the esophagus characterized by incomplete relaxation of the lower esophageal sphincter, resulting in obstruction. Aperistalsis and dilation of the esophageal hody occurs later, contributing to the esophageal dysfunction. Gastrointestinal bleeding in acbalasia is an infrequent complication usually caused by stasis ulcer, esophageal varices, carcinoma, or pneumatic dilation of the sphincter. We describe here a patient with longstanding achalasia who bled vigorously from a proximal esopbageal site that can be identified as arterial bleeding by endoscopy. Subsequent esopbageal resection allowed detailed histological and immunohistochemical examination, which revealed a vascular ectasia. This lesion was associated with an unusually rich network of nerve fibers containing calcitonin generelated peptide. Neuropeptide Y- and substance P- containing fibers were found to be decreased in this lesion as compared with controls. On the other hand vasoactive intestinal peptide- and nitric oxide synthasecontaining fibers appeared quantitatively similar to those of controls. Calcitonin gene-related peptide is known to be involved in angiogenesis and may bave played a causative role in the development of this lesion. Vascular ectasia may represent a hitherto unreported complication of achalasia.     

卡托普利对扩张型心肌病降钙素基因相关肽及淋巴细胞钙离子的影响

目的 :探讨卡托普利对扩张型心肌病 (DCM)患者血浆降钙素基因相关肽 (CGRP)及淋巴细胞 Ca2 的影响。方法 :采用放射免疫法和原子吸收光谱法分别测定卡托普利治疗 DCM患者前后的血浆 CGRP水平及淋巴细胞 Ca2 浓度。结果 :DCM患者血浆 CGRP水平及淋巴细胞 Ca2 浓度明显高于对照组 (均 P <0 .0 0 1 ) ,淋巴细胞 Ca2 浓度与射血分数呈负相关 (r=- 0 .51 ,P <0 .0 5) ,与血浆 CGRP水平呈正相关 (r =0 .58,P <0 .0 0 1 ) ,卡托普利组治疗后血浆 CGRP水平及淋巴细胞 Ca2 浓度明显降低 ,与治疗前比较 ,均 P <0 .0 0 1 ;而常规治疗组治疗前后心功能、血浆 CGRP水平及淋巴细胞 Ca2 浓度无明显变化。结论 :心肌细胞 Ca2 超负荷是 DCM心功能损害的重要原因 ,抑制心肌细胞 Ca2 超负荷可能是卡托普利治疗 DCM重要机制之一     

Calcitonin gene-related peptide in the hepatic and splanchnic vascular systems of the rat

The distribution and origin of calcitonin gene-related peptide immunoreactive structures in hepatic and splanchnic vasculature of the rat were investigated by the immunofluorescent technique. Calcitonin gene-related peptide immunoreactive fibers were dissociated from compact thick calcitonin gene-related peptide immunoreactive fiber bundles located around the hepatic and splanchnic vasculature, and reached the tunica adventitia of the vasculature. Some fibers penetrated further into the tunica media of the vasculature. In the tunica adventitia of the vasculature, calcitonin gene-related peptide immunoreactive fibers formed a fine meshwork which could be traced to the branches and arterioles. In the liver, calcitonin gene-related peptide immunoreactive innervation extended from porta hepatis to portal triads, running with branches of the hepatic artery and portal vein. Some calcitonin gene-related peptide immunoreactive fibers separated from thinner calcitonin gene-related peptide immunoreactive fiber bundles in portal triads and coursed into the immediately adjacent parenchyma. In the dorsal spinal ganglia (T8-10, L1), calcitonin gene-related peptide immunoreactive cells accounted for about 60% of the total number of ganglion cells. Bilateral vagotomy just below the diaphragm or bilateral transection of the greater splanchnic nerve resulted in a marked decrease of the number of calcitonin gene-related peptide immunoreactive fibers in the vasculature, whereas a combined operation was followed by the complete depletion of calcitonin gene-related peptide immunoreactive fibers. These results indicate that calcitonin gene-related peptide immunoreactive innervation in hepatic and splanchnic vasculature may have a dual origin. In conclusion, the widespread distribution of calcitonin gene-related peptide immunoreactive fibers in the hepatic and splanchnic vasculature suggest that calcitonin gene-related peptide innervation may be involved in sensory transmission.     

降钙素基因相关肽免疫反应纤维在大鼠心肌内的分布和密度

目的 观察降钙素基因相关肽免疫反应(CGRP-IR)纤维在大鼠心肌内的分布和密度，为心肌病的研究奠定形态学基础。方法 用免疫组织化学法显示大鼠心肌内的CGRP-IR纤维，计算其分布密度。结果在心房肌内和心室肌内均可见CGRP-IR纤维分布于心肌纤维之间，心房肌内CGRP-IR纤维的面积密度和数量密度均高于心室肌。结论 CGRP-IR纤维在大鼠心肌内分布广泛，其分布密度存在部位差异。     

Distribution and characterization of calcitonin gene-related peptide immunoreactivity in the digestive system of normal and capsaicin-treated rats

The distribution and characterization of calcitonin gene-related peptide immunoreactivity in the digestive system of normal, capsaicin-treated, and littermate control rats were studied by radioimmunoassay, chromatography, and immunohistochemistry. The highest concentrations of calcitonin gene-related peptide immunoreactivity were found in the stomach (45 +/- 2.8 pmol/g wet wt, nonsecretory region; 38.7 +/- 4.4 pmol/g wet wt, secretory region) and rectum (30.9 +/- 1.6 pmol/g wet wt). Significant amounts of peptide were also found in the other regions of the gut and in the pancreas. Neonatal treatment with capsaicin, which causes a permanent degeneration of most of the small-diameter sensory neurons, reduced calcitonin gene-related peptide content by greater than 95% in the esophagus and stomach, by 60% in the pancreas, and by less than 50% in the intestine, when compared with littermate controls. Separation of extracts from the gut, pancreas, and brain by chromatography gave major peaks corresponding to the predicted rat calcitonin gene-related peptide and small unidentified peaks, which presumably arise from metabolism of the peptide. Immunohistochemical studies demonstrated that in the esophagus and stomach, calcitonin gene-related peptide immunoreactivity is restricted to nerve fibers, whereas in the intestine it is localized in both nerve fibers and enteric ganglion cells. In capsaicin-treated rats there was a virtually complete elimination of calcitonin gene-related peptide immunoreactive fibers innervating the esophagus and stomach, whereas in the small and large intestine there was a dramatic reduction and often a complete elimination of those associated with blood vessels and a slighter reduction of the nonvascular immunoreactive fibers. The results of this study indicate that calcitonin gene-related peptide immunoreactive nerve fibers innervating the rat digestive system originate from both intrinsic (enteric) and extrinsic (presumably sensory) sources and that both the intrinsic and extrinsic components appear to contain a substance that corresponds to the predicted calcitonin gene-related peptide.     

Effects of lipids on the functional and metabolic recovery from global myocardial stunning in isolated rabbit hearts

OBJECTIVES: High concentrations of free fatty acids may increase myocardial ischaemic damage. However, the administration of lipid emulsions during reperfusion improves the functional recovery of stunned myocardium. From this apparent controversy we hypothesise that the effect of lipids is related to the time of its administration: we compared the effects of pre- and post-ischaemic administration of Intralipid((R)) on stunned myocardium. We also examined the role of fatty acids and phospholipids, respectively, in the effect of lipid emulsions on stunned myocardium. METHODS: Myocardial stunning was produced by 15 min of ischaemia and 90 min of reperfusion in isolated blood perfused rabbit hearts. Intralipid((R)) was administered either prior to ischaemia or during reperfusion. Left ventricular pressure (LVP) and its first derivative (LVdP/dt) were measured to assess functional recovery. High energy phosphates were measured with HPLC. The effects of linoleic acid, phosphatidylcholine and their combination were also studied. RESULTS: Only when Intralipid((R)) was administered during reperfusion, it improved recovery from contractile function and increased high energy phosphate content in globally stunned myocardium. Both linoleic acid and phosphatidylcholine significantly improved myocardial function in stunned myocardium. CONCLUSIONS: The effect of lipids on the contractile performance and metabolic state of stunned myocardium depends mainly on the timing of its administration with regard to the ischaemia/reperfusion event. Both free fatty acids and phospholipids contribute to the beneficial effect of lipid emulsions on functional recovery of stunned myocardium.     

Pathophysiology and pathogenesis of contractile failure in stunned myocardium

To investigate excitation-contraction coupling in stunned myocardium, intracellular free calcium concentration [( Ca2+]i) was measured before and after ischemia in perfused hearts using gated 19F NMR and the Ca2+ indicator 5F-BAPTA. Maximal Ca(2+)-activated force was also measured in parallel experiments. Stunned myocardium was created by reperfusion after 15 min global ischemia at 37 degrees C in isolated ferret hearts. In stunned myocardium, peak [Ca2+]i was paradoxically higher than that in control, but maximal Ca(2+)-activated pressure was lower in stunned hearts. These results indicate that contractile failure in stunned myocardium is due to a decrease in the myofilament sensitivity to Ca2+ as well as to a decrease in maximal Ca(2+)-activated force; failure of activator Ca2+ delivery cannot be implicated. The role of intracellular calcium overload in the pathogenesis of stunned myocardium was also investigated. Time-averaged 19F NMR measurements directly revealed the increase in [Ca2+]i during ischemia and in the early phase of reperfusion. The strategies to prevent Ca overload during reperfusion with modified reperfusate succeeded in preserving contractile function. Transient Ca overload without ischemia induced by different causes, i.e., high [Ca]0 perfusion, ventricular fibrillation or treatment with adriamycin, also produced contractile dysfunction that outlasted the interventions themselves. Thus, we propose that transient Ca overload during ischemia and early reperfusion initiates long-lasting contractile dysfunction in stunned myocardium.     

Stunned myocardium and sympathetic denervation--clinical assessment using MIBG scintigraphy

To evaluate the clinical relationship between stunned myocardium and the sympathetic nervous system, 6 patients who had stunned myocardium accompanied by T wave inversion underwent simultaneous 123I-metaiodobenzyl guanidine (MIBG) scintigraphy and thallium scintigraphy. All patients showed abnormal filling defects on the MIBG scintigrams in the areas with stunned myocardium but the thallium scintigrams were almost normal. The extent of the defects in these 6 patients was determined on the MIBG scintigrams using a Bull's eye display. The defects were found to be larger than those in 4 patients with angina pectoris, and those in 4 patients who had previously shown T wave inversion but had a normal electrocardiogram at the time of examination. Thus, it is suggested that sympathetic denervation is one of the mechanisms causing stunned myocardium.