神经肽Y对大鼠血管平滑肌细胞内钙离子浓度的影响

目的 应用激光共聚焦显微镜动态观察神经肽Y(NPY)受体激动剂[Leu31, Pro34]-NPY干预大鼠血管平滑肌细胞(VSMC)产生的细胞内钙离子([Ca2 ]i)浓度变化,以确定NPY对大鼠VSMC[Ca2 ]i浓度的影响.方法 用分散细胞培养法培养VSMC.用10 μmol/L的Fluo-3孵育细胞.使其与细胞内的游离[Ca2 ]i相结合,从而能够在525 nm的激发光激发下产生荧光.分别用正常细胞外液、无钙细胞外液稀释的10-6 mol/L[Leu31, Pro34 ]-NPY、1 μmol/L硝苯地平、10-6 mol/L[Leu31, Pro34 ]-NPY 1 μmol/L硝苯地平灌流细胞,同时用激光共聚焦显微镜扫描、记录图像.用Leica图像分析系统分析图像,定量分析不同情况下的[Ca2 ]i荧光强度.结果 正常细胞外液灌流时[Ca2 ]i变化不明显,最高值为57.3±3.1,最低值为53.7±2.9,无明显差异(P＞0.05). [Leu31, Pro34]-NPY干预细胞时,[Ca2 ]i明显地升高,随后又开始下降.最高值为86.4±2.7,最低值为58.1±3.0,有非常显著差异(P＜0.01).无钙细胞外液 [Leu31, Pro34 ]-NPY灌流VSMC时[Ca2 ]i浓度变化不明显,最高值为52.5±3.5,最低值为48.0±1.2(P＞0.05);[Leu31, Pro34 ]-NPY 硝苯地平组[Ca2 ]i变化不明显,最高值为52.6±2.1,最低值为51.7±2.7,差异无统计学意义(P＞0.05).硝苯地平组[Ca2 ]i呈下降趋势,最高值为52.8±2.2,最低值为37.5±2.1,有非常显著差异(P＜0.01).结论 NPY能够通过NPY-Y1受体使细胞内的[Ca2 ]i浓度升高,这种升高是依赖于细胞外的[Ca2 ]i由L型[Ca2 ]i通道进入细胞的.     

自发性高血压大鼠神经肽Y-Y1受体在脑血管平滑肌细胞增殖中的作用

目的探讨神经肽Y-Y1受体激动剂[Leu31,Pro34]-NPY对体外培养的自发性高血压大鼠(SHR)脑血管平滑肌细胞促增殖作用的影响.方法应用植块法体外培养SHR大脑动脉血管平滑肌细胞,分别用正常细胞外液、10-5或10-8NPY,10-6硝苯地平,10-6硝苯地平 10-6mol/L NPY加入培养液培养细胞.用免疫荧光法检测平滑肌培养细胞中增殖核抗原(PCNA)的表达,用流式细胞仪对平滑肌细胞的S期细胞进行检测分析.结果随着[Leu31,Pro34]-NPY浓度的增加,平滑肌细胞PCNA阳性率递增,与对照组相比其阳性率有明显差别(P＜0.001).硝苯地平明显抑制PCNA阳性率.与硝苯地平 10-6 mol/L NPY相比,PCNA阳性率有显著差异(5.1±0.5 vs 17.7±2.5,P=0.009).随着NPY浓度增加,血管平滑肌细胞S期百分数也是增加的,10-5、10-6mol/L NPY组与对照组相比有差别(P＜0.05).硝苯地平组与硝苯地平 10-6mol/L NPY比较,S期细胞百分数有显著差异(P=0.0023).结论NPY-Y1受体激动剂对血管平滑肌细胞具有浓度依赖性的PCNA表达增强作用,能够提高血管平滑肌细胞S期比例.L型钙离子拮抗剂能够减弱NPY-Y1受体激动剂的促平滑肌细胞增殖作用.     

自发性高血压大鼠神经肽Y-Y1受体在脑血管平滑肌细胞增殖中的作用

目的探讨神经肽Y-Y1受体激动剂[Leu~(31),Pro~(34)]-NPY对体外培养的自发性高血压大鼠(SHR)脑血管平滑肌细胞促增殖作用的影响。方法应用植块法体外培养SHR大脑动脉血管平滑肌细胞,分别用正常细胞外液、10~(-5)或10~(-8)NPY,10~(-6)硝苯地平,10~(-6)硝苯地平 10~(-6)mol/L NPY加入培养液培养细胞。用免疫荧光法检测平滑肌培养细胞中增殖核抗原(PCNA)的表达,用流式细胞仪对平滑肌细胞的S期细胞进行检测分析。结果随着[Leu~(31),Pro~(34)]-NPY浓度的增加,平滑肌细胞PCNA阳性率递增,与对照组相比其阳性率有明显差别(P<0·001)。硝苯地平明显抑制PCNA阳性率。与硝苯地平 10~(-6)mol/L NPY相比,PCNA阳性率有显著差异(5·1±0·5vs17·7±2·5,P=0·009)。随着NPY浓度增加,血管平滑肌细胞S期百分数也是增加的,10~(-5)、10~(-6)mol/L NPY组与对照组相比有差别(P<0·05)。硝苯地平组与硝苯地平 10~(-6)mol/L NPY比较,S期细胞百分数有显著差异(P=0·0023)。结论NPY-Y1受体激动剂对血管平滑肌细胞具有浓度依赖性的PCNA表达增强作用,能够提高血管平滑肌细胞S期比例。L型钙离子拮抗剂能够减弱NPY-Y1受体激动剂的促平滑肌细胞增殖作用。     

黄芪抑制去甲肾上腺素诱导大鼠心肌细胞内游离钙含量升高的作用

目的 :研究黄芪对去甲肾上腺素 (NE)诱导大鼠心肌细胞内游离钙 [Ca2 + ]i 含量升高的作用。方法 :应用AR CM MIC阳离子检测系统 ,采用Ca2 + 指示剂Fura 2 /AM检测培养新生大鼠单个心肌细胞内游离钙的浓度 ,并观察Am对NE诱导大鼠心肌细胞内 [Ca2 + ]i 升高的影响。结果 :当细胞外Ca2 + 浓度为 1 .3mmol/L时 ,大鼠心肌细胞 [Ca2 + ]i 为 ( 97.1 1 +8.2 1 )nmol/L ,给予 1 0 -5mol/L的NE ,可使大鼠心肌细胞 [Ca2 + ]i 增至 ( 2 91 .73± 1 5 .0 3 )nmol/L,而经黄芪处理的大鼠心肌细胞的 [Ca2 + ]i则由 ( 95 .98±8.1 4)nmol/L增至 ( 1 1 4.2 8± 9.2 9)nmol/L。结论 :NE能诱导大鼠心肌细胞内游离钙含量明显升高 ,黄芪对心肌细胞静息 [Ca2 + ]i无明显影响 ,但能抑制NE诱导大鼠心肌细胞 [Ca2 + ]i 升高     

降钙素基因相关肽对血管升压素缩血管效应的影响

目的 :探讨降钙素基因相关肽 （CGRP）对血管升压素 （AVP）缩血管效应的影响。方法 :培养大鼠主动脉血管平滑肌细胞 （VSMC） ,给予 AVP和 CGRP干预 ,检测 VSMC的细胞内游离钙离子浓度 （[Ca2 + ] i）。结果 :AVP增加VSMC的 [Ca2 + ] i,而 CGRP明显抑制 AVP的这一作用。结论 :CGRP可能通过抑制 VSMC的 [Ca2 + ] i的升高而发挥其对 AVP所致缩血管效应的抑制作用     

急性缺氧对环匹阿尼酸诱导的大鼠远端肺静脉平滑肌细胞内钙浓度升高的影响

目的:研究急性缺氧对Ca2+-ATPase抑制剂——环匹阿尼酸(CPA)诱导的大鼠远端肺静脉平滑肌细胞(PVSMC)内钙浓度([Ca2+]i)升高的影响及机制。方法:选用6只雄性SD大鼠(体重200~250 g),培养大鼠PVSMC,运用荧光显微镜和细胞内钙浓度检测系统观测CPA及急性缺氧(4%O2)对PVSMC的[Ca2+]i影响及钙池操纵性钙通道(SOCC)阻断剂氯化镍(Ni Cl2)和SKF96365的干预作用。结果:含5μmol/L硝苯地平(电压依赖钙通道阻断剂)的无钙Krebs溶液孵育PVSMC,10μmol/L CPA使PVSMC的[Ca2+]i小幅度升高,急性缺氧能使[Ca2+]i升高幅度增加;恢复细胞外Ca2+至2.5 mmol/L后,10μmol/L CPA使[Ca2+]i显著升高,急性缺氧可导致CPA诱导的[Ca2+]i升高显著增强;SOCC阻断剂Ni Cl2(500μmol/L)和SKF96365(50μmol/L)均能明显抑制急性缺氧条件下CPA诱导的[Ca2+]i升高,但对高钾(60 mmol/L KCl)Krebs溶液引起的[Ca2+]i反应无影响。结论:急性缺氧能够使CPA诱导的大鼠远端PVSMC的[Ca2+]i升高增强,[Ca2+]i升高可被SOCC阻断剂阻断,提示急性缺氧能够增强SOCC活性,使细胞外Ca2+通过SOCC内流增强,从而导致大鼠远端PVSMC的[Ca2+]i升高。     

Fura-2/AM荧光法测定日本血吸虫培养细胞内游离钙离子浓度

目的　研究用钙荧光探剂 (Fura- 2 / AM)检测静息状态下日本血吸虫培养细胞内的游离钙离子浓度 ([Ca2 + ]i) ,以及β-巯基乙醇对培养细胞 [Ca2 + ]i的影响。方法　将 18d虫龄的日本血吸虫童虫制成细胞悬液 ,贴壁法接种于 30 ml培养瓶中 ,培养液为 RPMI- 16 4 0含 2 0 %小牛血清及常量抗生素。在培养的 0 - 3d,制备日本血吸虫细胞悬液 ,采用 Fura- 2 / AM钙荧光探剂测定正常静息状态下及加入β-巯基乙醇后日本血吸虫培养细胞内的 [Ca2 + ]i。结果　正常静息状态下 ,培养 0 d的日本血吸虫培养细胞内的 [Ca2 + ]i为 188.2 nmol/ L ;培养 1- 3d的 [Ca2 + ]i两两比较差异无显著性(P>0 .0 5 ) ,它们的平均值为 (187.0± 10 .7) nm ol/ L ,与培养 0 d的 [Ca2 + ]i比较 ,差异也无显著性(P>0 .0 5 )。β-巯基乙醇可使日本血吸虫培养细胞内 [Ca2 + ]i明显升高 (P<0 .0 1) ,并随浓度的增加而升高。结论　培养 1- 3d,静息状态下日本血吸虫培养细胞内的 [Ca2 + ]i比较稳定 ,β-巯基乙醇能影响日本血吸虫培养细胞内的 [Ca2 + ]i     

Leptin transiently antagonizes ghrelin and long-lastingly orexin in regulation of Ca2＋ signaling in neuropeptide Y neurons of the arcuate nucleus

AIM： To explore the mechanism for interactions of leptin with ghrelin and orexin in the arcuate nucleus （ARC） activating neuropeptide Y （NPY） neurons during physiological regulation of feeding, METHODS： Single neurons from ARC of adult rats with matured feeding function were isolated. [Ca2＋]i was measured to monitore their activities. The time course of leptin effects on ghrelin-induced versus orexin-induced [Ca2＋]i increases in NPY neurons was studied. RESULTS： Administration of ghrelin or orexin-A at 101~ mol/L increased cytosolic Ca2~ concentration （[Ca2＋]~） in NPY neurons isolated from the ARC of adult rats. Upon administration of leptin at 10^-14-10^-12 mol/L, ghrelin-induced [Ca2＋]i increases were initially （〈 10 min） inhibited but later restored, exhibiting a transient pattern of inhibition. In contrast, orexin-induced [Ca2＋]i increases were inhibited by leptin in a long- lasting manner. Furthermore, a prior administration of leptin inhibited orexin action but not ghrelin action to increase ICa 2＋li, CONCLUSION： Leptin counteracted ghrelin effects transiently and orexin effects long-lastingly in NPY neurons. The transient property with which leptin counteracts ghrelin action in NPY neurons may allow the fasting-associated increase in ghrelin levels to activate NPY neurons in the presence of physiological leptin and to stimulate feeding.     

营养性低钙与慢性氟中毒大鼠细胞内钙超载相关性研究

目的探讨营养性低钙与慢性氟中毒细胞内钙超载的相关性。方法采用饮水加入氟化钠进行大鼠染毒实验,配以低钙饮食,应用Fura-2／AM荧光指示剂标记单细胞,显微荧光标记测定慢性氟中毒大鼠肝、肾、脑细胞内游离钙([Ca2 ]i)的变化。结果氟可刺激细胞内[Ca2 ]i水平增高,正常饮食投氟组肝、肾、脑细胞内[Ca2 ]i水平分别为(124．11±11.35)、(127．20±14．40)、(136．75±15．91)nmol／L,与正常饮食对照组(86．37±27．55)、(95．65±8．75)、(99．20±12．89)nmol／L比较,差异有统计学意义(t=4．001、4．706、4．953,P< 0．05);低钙饮食 氟组肝、肾、脑细胞内[Ca2 ]i水平[(151．57±19．61)、(162．62±31．41)、(158．79±27．65) nmol／L]与低钙饮食对照组[(103．17±14．29)、(103．59±11．40)、(115．26±30．91)nmol／L]比较,差异有统计学意义(t=4．789,P<0．05;t=8．581、6．003,P<0．01);低钙饮食 氟组骨病变明显:细胞内[Ca2 ]i水平与血清离子钙(i[Ca])、总钙(t[Ca])呈负相关(r=-0．59)。结论慢性氟中毒可致机体组织细胞钙超载,低钙可加重氟中毒时的细胞内钙超载,提示细胞内钙超载可能参与氟骨症发病机制并起着重要作用。     

钙神经素信号途径在前列腺素F2α诱导乳鼠心肌细胞肥大中的作用

目的:探讨Ca2 /钙调蛋白(CAM)依赖的钙神经素(CAN)信号途径在前列腺索F2α(PGF2α)诱导心肌细胞肥大中的作用.方法:用PGF2α(1 μmol/L,0.1μmol/L,0.01 μmol/L,0.001μmol/L)刺激Wistar乳鼠心肌细胞,并用CaN特异性抑制剂环胞素A(CsA)加以干预.以心肌细胞总蛋白含量和心肌细胞面积作为反映心肌肥大的指标,用免疫印迹(Western-blot)测定心肌细胞内CaN-α蛋白表达,用Fura 2/AM为荧光指示剂测定心肌细胞内游离钙浓度([Ca2 ]i).结果:除0.001μmol/L浓度的PGF2α外,1.0μmol/L,0.1μmol/L,0.01μmol/L的PGF2α均可明显提高心肌细胞内蛋白质浓度,而其中0.1μmol/L PGF2α作用最显著,增加了(67.78±11.19)%,并且使乳鼠心肌细胞面积增加了(68.94±1.52)%.0.1μmol/L PGF2α刺激的心肌细胞CaN-α蛋白表达增加和[Ca2 ]i提高,并且加入CsA可阻断0.1μmol/L PGF2α诱导的乳鼠心肌细胞肥大效应.结论:PGF2α可能通过Ca2 /CaM-CaN信号途径刺激乳鼠心肌细胞发生肥大.     

青少年高血压的研究进展

随着人们生活和行为方式的改变,高血压发病明显呈年轻化趋势。在青少年时期识别高血压病高危人群有助于早期进行有效干预和治疗,降低未来高血压的发生率及其严重性。现试从青少年高血压的诊断、发病因素、特点、治疗策略等方面的研究进展作一综述。     

Morbidity in cardiovascular diseases in immigrants in Sweden

INTRODUCTION: Although immigration to Sweden has increased in the last few decades, the incidence rates of cardiovascular disease and coronary heart disease in immigrants are unknown. The aim of the present study is to estimate whether place of birth affects the incidence rates of cardiovascular disease and coronary heart disease. MATERIAL AND METHODS: The study was designed as a follow-up study on morbidity in cardiovascular disease and coronary heart disease between 1 January 1997 and 31 December 1998, including three and a half million persons with age range 35-64 years, of whom 550 000 were born abroad, from the database MigMed consisting of the whole Swedish population. Incidence rates and relative risks were estimated by indirect standardization and a proportional hazard model. RESULTS: The age-adjusted risk of coronary heart disease was higher in most foreign-born groups than in Swedes. For example, in nine of 12 male groups, the relative risks varied between 1.1 and 2.2, and in seven of 12 female groups, the relative risks varied between 1.4 and 2.5. When also adjusting for level of education and employment status, the risks were still high, but on a lower level. CONCLUSIONS: Foreign-born people possess an over-risk of cardiovascular or coronary heart disease(CVD/CHD) compared with Swedish-born persons, also when level of education and employment status are taken into account.     

C·肉毒杀鼠素(冻干剂)应用研究

目的为研究C·肉毒杀鼠索对杀灭达乌尔黄鼠(简称黄鼠)的大面积应用情况和对家畜、家禽的毒害作用,进行了C·肉毒杀鼠素的应用研究.方法大面积投毒采用ES-2药饵撒播机[1],间隔约80m进行条投.羊、鸡采用直接灌胃.结果大面积应用的灭鼠率为83.72%.对羊、鸡最高剂量分别为500万MLD、150万MLD,均未出现中毒现象.结论 C·肉毒杀鼠素是较为理想的草原大面积杀灭黄鼠的理想、首选药物.     

高龄老年高血压的临床研究进展

随着社会老龄化进程的加快,80岁以上高龄老人的绝对数量以及占总人口的比例均在增加,如何提高高龄老年高血压的防治水平备受关注。现试从高龄老年高血压的临床特点、治疗策略等方面的临床研究进展作一综述。     

中国老年性耳聋的研究与干预进展

老年性耳聋已成为影响我国老年人生活质量的最主要的慢性病之一。助听器是目前帮助听力损失的老年人克服交流障碍的主要手段。在我国,数字助听器已逐渐取代模拟助听器并且体现出了更好的效果。但是老年听力损失患者中使用助听器的比例仍然很小。人工耳蜗植入也已被应用在老年患者中。我国针对老年人的听力康复服务还有较长的路要走。     

Sex Differences in Ethanol-induced Dopamine Release in Nucleus Accumbens and in Ethanol Consumption in Rats

In vivo microdialysis was used to examine changes in nucleus accumbens and striatal dopamine, dihydrophenylacetic acid (DO-PAC), and homovanillic acid (HVA) following acute administration of ethanol (0.0, 0.25, 0.5, 1.0, or 2.0 g/kg) in male and female Long-Evans rats. Following dialysis, rats were trained to bar-press for oral ethanol reinforcement. In nucleus accumbens, females showed significant increases in extracellular dopamine following 0.25 or 0.5 g/ kg ethanol, but did not show significant increases over baseline at the higher doses. Males showed slight increases in dopamine at the lower doses and decreased dopamine at 2.0 g/kg. In striatum, both sexes showed increased dopamine at the lower doses and decreased dopamine at 2.0 g/kg. There were slight increases in nucleus accumbens DOPAC and HVA at some doses in both sexes, but no changes in striatal metabolite levels. In addition to showing increased responsiveness to ethanol-induced mesolimbic dopamine stimulation, females consumed more ethanol than males during behavioral testing. The pattern of both greater ethanol-induced nucleus accumbens dopamine release and greater ethanol consumption in females supports the hypothesis that ethanol reward is mediated, at least in part, by the mesolimbic dopamine system.     

Epstein-Barr virus in malignancies in renal transplant recipients in Japan

A role for Epstein-Barr virus (EBV) in the development of malignancies including lymphomas, and carcinoma of the stomach, nasopharynx, thymus and salivary gland is suggested. It is indicated that EBV evokes polyclonal-B-cell-proliferative diseases in immunocompromised hosts, such as transplant patients, which results in monoclonal malignant lymphomas. The suppression of immune functions in these patients is thought to lead to incomplete elimination of the cells expressing EBV latent infection genes. To examine the etiological role of EBV in the development of malignancies following renal transplant in Japan, 42 malignancies in 1744 cases of renal transplant were studied for the presence and type of EBV. The polymerase chain reaction revealed that 5 malignancies were positive for EBV, all type A: 2 of 2 cases of non-Hodgkin's lymphoma (NHL), 2 of 8 cases of gastric adenocarcinoma of the common type, and 1 of 2 cases of gastric plasmacytoma. In situ hybridization revealed positive signals in the nucleus of tumor cells in 2 cases of NHL and 1 of plasmacytoma. Positive signals were found in the small lymphoid cells but not in the tumor cells in 2 cases of gastric carcinoma. On the basis of these findings, a role for EBV in the development of malignancies in renal transplant patients is unlikely except for lymphoid neoplasias.Abbreviations PCB polymerase chain reaction - EBV Epstein-Barr virus - NHL non-Hodgkin's lymphoma     

糖尿病大鼠心室肌细胞钾通道的改变

目的研究糖尿病大鼠心室肌细胞钾通道的改变及增加葡萄糖代谢对其的作用。方法取体重150～200g的雄性SpragueDawley大鼠,腹腔注射链脲菌素(STZ)建立糖尿病模型,采用酶解法获得单个心室肌细胞,应用膜片钳全细胞记录技术记录钾电流。结果糖尿病大鼠心室肌细胞瞬间外向性钾流(Ito)密度较对照组显著降低[ 60mV时,分别为(15.90±1.19)pA/pF(n=25)和(28.55±0.97)pA/pF(n=12),P<0.001];分别用100nmol/L胰岛素及1.5mmol/L二氯乙酸在体外预处理心室肌细胞4～5h和3～4h使Ito密度恢复至对照组水平[ 60mV时,分别为(29.40±0.38)pA/pF(n=20)和(27.35±0.97)pA/pF(n=12)]。结论糖尿病大鼠心室肌细胞钾通道功能发生改变,增加葡萄糖代谢可逆转这一改变,提示葡萄糖代谢与Ito功能间存在一定关系。