普罗布考对动脉粥样硬化大鼠平滑肌细胞凋亡及p53和Fas蛋白表达的影响

目的探讨普罗布考防治动脉粥样硬化（AS）的机制。方法选用雄性大鼠,复制大鼠AS模型,随机分为动脉粥样硬化模型组、普罗布考组和正常对照组。大鼠造模成功后给予普罗布考治疗,6周后处死大鼠,采用流式细胞术检测平滑肌细胞凋亡率及凋亡相关基因p53和Fas蛋白的表达。结果模型组大鼠血管平滑肌细胞凋亡率明显高于对照组（P〈0．05）,p53和Fas蛋白的表达增强（P％0．05）,主动脉壁可肉眼观测典型斑块。普罗布考组大鼠平滑肌细胞的凋亡率明显低于模型组（P〈0．05）,p53和Fas蛋白表达下调（P〈0．05）,主动脉斑块面积较模型组减小明显。结论普罗布考通过调节p53和Fas蛋白表达来调节AS大鼠平滑肌细胞的凋亡。     

大鼠心肌肥厚时左心室心房钠尿肽基因的转录

本工作运用ANF放射免疫测定和ANF核酸分子杂交的方法,测定了大鼠腹主动脉部分结扎引起心肌肥厚时血浆ANF及左心室ANF mRNA水平的变化。结果表明,心肌肥厚大鼠的血浆ANF及左心室ANF mRNA水平都明显升高,提示大鼠心肌肥厚可以促进其左心室ANF基因的转录和表达。此外,本工作还证明细胞内钙离子调节剂牛磺酸和血管松弛剂肼苯哒嗪可以分别促进和抑制大鼠心肌肥厚诱导的左心室ANF基因的转录和表达。     

围术期应用肼苯哒嗪预防疝修补术后慢性疼痛的研究

目的：探讨肼苯哒嗪对疝修补术后患者慢性疼痛发生率的影响。方法：选择单侧腹股沟疝患者48人,随机分为实验组（T组）和对照组（c组）各24例,在术前3天至术后4天共7天内接受药物或安慰荆治疗。其中T组每日两次,每次10mg肼苯哒嗪,C组同样方式给予安慰剂。测量患者术后4天内的平均动脉）Z（MAP）、心率（HR）、NRS急性疼痛评分以及术后3月、6月S—LANSS慢性疼痛评分。结果：T组患者术后MAP明显低于C组（P〈0．05）,HR两组相比无差异（P〉0．05）;两组患者术后急性疼痛程度相比无统计学差别（P〉0．05）;T组患者在术后3月和6月慢性疼痛发生率明显低于C组（P〈0．05）。结论：常规剂量肼苯哒嗪可以用于术后患者,对疝修补术后急性疼痛无影响,但可以明显降低术后慢性疼痛的发生率。     

NO前体对高肺血流时肺动脉胱硫醚-γ-裂解酶基因表达的调节作用

目的：研究一氧化氮（NO）前体L-精氨酸（L—Arg）对高肺血流时肺动脉胱硫醚-γ-裂解酶（内源性硫化氢生成酶）的调节作用,以探讨NO体系对高肺血流肺动脉高压及肺血管结构重建调节作用的机制。方法：30只雄性SD大鼠随机分为对照组,分流组和分流＋L—Arg组。对后两组大鼠行腹主动脉-下腔静脉分流术。观察术后11周大鼠肺动脉平均压（mPAP）、右心室肥厚和肺动脉相对中膜面积的改变,用竞争逆转录聚合酶链反应（RT-PCR）对肺组织CSEmRNA表达进行定量分析,同时用化学法测定肺组织硫化氢产出率。结果：分流组大鼠mPAP及肺动脉相对中膜面积明显高于对照组（P〈0．01）,而分流＋L—Arg组大鼠mPAP及肺动脉相对中膜面积明显低于分流组（P〈0．01）。分流组CSEmRNA表达与对照组相比明显降低（P〈0．01）,而分流组＋L—Arg组CSEmRNA表达又明显高于分流组（P〈0．05）：分流组大鼠肺组织硫化氢产出率明显低于对照组（P〈0．01）,而分流组＋L—Arg组大鼠肺组织硫化氢产出率及血浆硫化氢含量明显高于分流组（P〈0．01）。结论：高肺血流可致肺动脉CSEmRNA下调,外源性NO能够缓解CSEmRNA的改变,从而对高肺血流所致肺血管结构重建和肺动脉高压起调节作用。     

筛选大鼠血管钙化消退差异表达基因及初步分析

目的：探讨大鼠在血管钙化消退过程中基因表达的变化。方法：选取6周龄SPF级雄性SD大鼠24只,随机分成3组（n=8）,分别为对照组、钙化组和消退组。钙化组和消退组制作血管钙化模型（vitamin D3 plus nieotine,VDN）,对照组生理盐水和花生油灌胃。钙化组和对照组于实验第8周处死,消退组继续饲养至16周处死,测定各组大鼠动脉组织钙含量并作病理检查。采用抑制性消减杂交方法将消退组和钙化组大鼠血管cDNA作差减杂交,分离消退组较钙化组高表达或低表达基因的cDNA片段,建立消退组和钙化组的差异表达文库,扩增、鉴定文库并测序阳性克隆,BLAST比对测序序列。随机挑选4个基因进行RT-PCR验证和DNA条带半定量分析。结果：①血管组织钙含量测定显示钙化组（（15．34±2．51）mg／g）较对照组（（5．20±0．75）mg／g）血管组织钙含量明显升高（P〈0．01）;消退组（（12．73±1．89）mg／g）较钙化组降低（P〈0．05）;②构建了血管钙化的差减文库,对所获阳性克隆进行测序和BLAST比对分析,获得28个表达上调基因和22个表达下调基因。RT-PCR验证示Prdx3,Ank2,Ror2,Abcel等基因在消退组和钙化组间差异表达,消退组较钙化组表达增高,平均约为后者的1．7倍。结论：VDN模型诱导的大鼠血管钙化可以发生主动消退。钙化消退过程中焦磷酸合成相关基因、谷氨酸信号相关基因、还原及凋亡调节基因表达上调,同时见较多骨化相关基因及氧化活性等基因表达下调。钙化抑制基因的表达增多而钙化促进基因表达的下降可能是钙化发生主动消退的内源性机制。     

运动训练对大鼠主动脉应力和NF-κB及c-fos表达的影响

目的：探讨不同强度运动训练对大鼠主动脉应力和NF-κB及c-fos表达的影响。方法：采用跑台训练方式,建立大鼠有氧运动和疲劳运动模型,运用免疫组织化学SABC法研究不同强度运动训练对大鼠主动脉VEC和VSMC中NF-κB及c-fos表达的影响。结果：胸主动脉血压的变化与对照组比较,有氧训练和疲劳训练均显著性升高（P〈0．05）,但有氧训练与疲劳训练之间差异不显著。与对照组比较,有氧训练大鼠VEC和VSMC中NF-κB和c-fos均显著性下调表达,胸主动脉张开角显著性升高（P〈0．05）,疲劳训练大鼠VEC和VSMC中NF-κB和c-fos均显著性上调表达（P〈0．05）。与有氧运动组比较,疲劳运动大鼠VEC和VSMC中NF-κB和c-fos表达与胸主动脉张开角升高均具有显著性差异（P〈0．05）。表明不同强度运动大鼠主动脉VEC中NF-κB和c-fos表达变化趋势不同,疲劳训练组表达的增加趋势更显著。结论：运动可引起大鼠主动脉VEC和VSMC NF-κB及c-fos表达的变化,且与运动强度关系密切。有氧训练引起的慢性剪切应力变化可使主动脉VEC和VSMC NF-κB加及c-fos显著性下调表达,与VEC和VSMC维持血管功能的稳态有关;疲劳训练可导致壁面摩擦剪切力、周向应力增加,过度的剪切力作用可引起主动脉VEC和VSMC NF-κB及c-fos显著性上调表达,血管张开角显著增加,血管发生非均匀生长,引起血管结构与功能的重塑。     

跑台运动和营养补充对大鼠血红素代谢限速酶和珠蛋白mRNA表达与活性水平的影响

目的：研究血红素代谢限速酶和珠蛋白代谢在运动性贫血发生机理中的功能和作用,及营养补充对运动性贫血防治效果的作用机制。方法：本实验对30只雄性Wistar大鼠进行等量随机分为3组（n=10）：对照组（C）、运动组（P）和运动＋营养组（G）。30m／min、0％坡度、每次1min为起始训练方式,前5周和后4周时训练时间的加速度为每次2min,训练频率为每天2次（前两周例外）。11周的跑台运动结束后应用RT-PCR和免疫组织化学的方法测试骨髓每氨基-γ酮戊酸合成酶（ALAs）、铁螯合酶（ferrochelatase）、α-珠蛋白、β-珠蛋白的基因表达和肝脏血红素氧舍酶-1（HO-1）的活性。结果：11周跑台运动可以增加大鼠肝脏HO-1的活性和骨髓β-珠蛋白的基因表达（P〈0．01,P〈0．05）,抗运动性贫血复合剂补充并不能改变大鼠运动后血红素代谢限速酶和珠蛋白基因表达和活性,且运动＋营养组大鼠肝脏HO-1活性水平显著高于对照组（P〈0．01）,即递增负荷跑台运动不能影响大鼠骨髓血红素合成酶和α-珠蛋白的基因表达,但能够影响大鼠肝脏血红素分解酶的活性水平和骨髓β-珠蛋白的基因表达。结论：肝脏HO-1活性水平的升高可能是运动性贫血表现出低Hb、RBC和Hct水平的原因之一。     

蒺藜皂苷对动脉粥样硬化大鼠动脉壁ICAM-1、VCAM-1、PPARα、PPARγ基因表达的影响

观察全草蒺藜皂苷（tribu saponin from Tribulus terrestris,STT）对实验性动脉粥样硬化（atherosclerosis,AS）大鼠动脉壁中ICAM-1、VCAM-1、PPARα和PPARγ基因表达的影响,以探讨STT抗AS的机制。应用高脂饲料饮食配合注射维生素D,建立SD大鼠AS模型,并设立正常组、模型组、辛伐他汀组和蒺藜皂苷低、中、高剂量组。采用半定量RT-PCR的方法检测各组动物动脉壁中ICAM-1、VCAM-1、PPARα和PPARγ基因的表达,分析造模及各给药大鼠ICAM-1、VCAM-1、PPARα和PPARγ基因表达的变化。与正常组相比,模型组ICAM-1和VCAM-1基因的表达量明显增加（P〈0．01）,而PPARα和PPARγ基因的表达量明显降低（P〈0．01）;与模型组相比,辛伐他汀及各STT药均能降低ICAM-1和VCAM-1基因的表达量（P〈0．01～P〈0．05）,并能增加PPARα和PPARγ基因的表达量（P〈0．01）。提示STT能下调实验性AS大鼠动脉壁ICAM-1和VCAM-1基因的表达及上调PPARα和PPARγ基因表达,这可能是STT抗AS的作用机制之一。     

缺血后处理对肺缺血／再灌注损伤的保护作用及其机制

目的：探讨缺血后处理（聃）是否通过抑制P38丝裂原活化蛋白激酶（P38MAPK）活化来减轻再灌注损伤肺细胞的凋亡。方法：雄性SD大鼠40只,随机分成5组（n=8）,即对照组（C组）、肺缺血／再灌注组（I／R组）、肺缺血／再灌注＋缺血后处理组（IPO组）、缺血后处理＋溶剂对照组（D组）、缺血后处理＋SB203580组（SB组）。各组分别于再灌注2h留取左肺组织,检测肺组织湿／干重比（W／D）和总肺含水量（TLW）;光镜观察肺组织形态学结构改变并进行肺组织损伤定量评估（IQA）;原住末端标记法（TUNEL）检测肺细胞凋亡情况并计算凋亡指数（AI）;RT-PCR和免疫组化法测定Bax、Bcl-2基因和蛋白的表达。结果：与C组相比,I／R组W／D、TLW、IQA和AI均显著升高（P〈0．05,P〈0．01）,肺组织结构发生明显损伤;Bcl-2、Bcl-2／Bax基因及蛋白表达明显降低,Bax基因及蛋白表达明显升高（P〈0．05,P〈0．01）;IPO组、D组、SB组与I／R组相比,w／D、TLW、IQA和AI均显著降低（P〈0．05,P〈0．01）,肺组织结构损伤情况有所改善;Bcl-2、Bcl-2／Bax基因及蛋白表达明显升高,Bax基因及蛋白表达明显降低（P〈0．05,P〈0．01）;D组与IPO组比较各项指标均无明显差异（均P〉0．05）;SB组与IPO组相比,肺组织W／D、TLW、IQA和AI均显著降低（P〈0．05,P〈0．01）,肺组织结构未见明显损伤;Bcl-2、Bcl-2／Bax基因及蛋白表达明显升高,Bax基因及蛋白表达明显降低（P〈0．05,P〈0．01）。结论：I／R通过激活P38MAPK导致大鼠肺泡结构严重破坏,肺内细胞大量凋亡;IPO可能是通过抑制P38MAPK通路的激活而减轻L／R损伤。     

NO和HIF-1α在大鼠低氧性肺动脉高压中的作用及相互关系

目的：探讨低氧诱导因子1α（HIF-1α）在慢性低氧高二氧化碳性大鼠肺动脉高压中的变化及其与一氧化氮（NO）的关系。方法：SD大鼠40只随机分为正常对照组（NC）、低氧高二氧化碳组（HH）、低氧高二氧化碳加L-精氨酸（L-Arg）脂质体组（HP）、低氧高二氧化碳加L-硝基-精氨酸甲酯（L-NAME）组（HM）。测定平均肺动脉压（mPAP）、右室／（左室＋室间隔）重量比[RV／（LV＋S）]、血浆和肺组织匀浆一氧化氮（NO）含量。免疫组织化学和原位杂交法检测肺细小动脉HIF-1α及HIF-1αmRNA、内皮结构型一氧化氮合酶（ecNCS）蛋白及其mRNA的表达。结果：①HH组mPAP、RV／（LV＋S）高于NC组（P〈0．05）,HP组低于HH组（P〈0．01）;HM组mPAP高于HH组（P〈0．05）,RV／（LV＋S）与HH组差异无显著性。②HH组血浆及肺组织匀浆NO含量低于NC组（P〈0．01）,HP组高于HH组（P〈0．01）;HM组血浆、肺组织匀浆NO含量与HH组差异无显著性。③HH组肺细小动脉HIF-1α蛋白及HIF-1αmRNA表达高于NC组（P〈0．01）,ecNOSmRNA的表达低于NC组（P〈0．01）;HP组肺细小动脉HIF-1α蛋白及HIF-1αmRNA表达低于HH（P〈0．01）,ecNOS蛋白和ecNOSmRNA的表达高于HH组（P〈0．01）;HM组肺细小动脉HIR-1α蛋白及mRNA表达高于HH组（P〈0．05）,ecNOS蛋白和mRNA的表达低于HH组（P〈0．05）。结论：HIF-1α参与了慢性低氧高二氧化碳性大鼠肺动脉高压的形成,NO可能部分通过影响HIF-1α的表达和／或活性而实现其肺血管保护效应。     

Mineralocorticoid receptor antagonism protects the aorta from vascular smooth muscle cell proliferation and collagen deposition in a rat model of adrenal aldosterone-producing adenoma

The number of patients with adrenal aldosterone-producing adenomas (APAs) has gradually increased. However, even after adenoma resection, some patients still suffer from high systolic blood pressure (SBP), which is possibly due to great arterial remodeling. Moreover, mineralocorticoid receptors (MRs) were found to be expressed in vascular smooth muscle cells (VSMCs). This study aims to determine whether MR antagonism protects the aorta from aldosterone-induced aortic remolding. Male rats were subcutaneously implanted with an osmotic minipumps and randomly divided into four groups: control; aldosterone (1 μg/h); aldosterone plus a specific MR antagonist, eplerenone (100 mg/kg/day); and aldosterone plus a vasodilator, hydralazine (25 mg/kg/day). After 8 weeks of infusion, aortic smooth muscle cell proliferation and collagen deposition, as well as the MDM2 and TGF-β1 expression levels in the aorta, were examined. Model rats with APAs were successfully constructed. Compared with the control rats, the model rats exhibited (1) marked SBP elevation, (2) no significant alteration in aortic morphology, (3) increased VSMC proliferation and MDM2 expression in the aorta, and (4) enhanced total collagen and collagen III depositions in the aorta, accompanied with up-regulated expression of TGF-β1. These effects were significantly inhibited by co-administration with eplerenone but not with hydralazine. These findings suggested that specific MR antagonism protects the aorta from aldosterone-induced VSMC proliferation and collagen deposition.     

Aldosterone induces a vascular inflammatory phenotype in the rat heart

Vascular inflammation was examined as a potential mechanism of aldosterone-mediated myocardial injury in uninephrectomized rats receiving 1% NaCl-0.3% KCl to drink for 1, 2, or 4 wk and 1) vehicle, 2) aldosterone infusion (0.75 microg/h), or 3) aldosterone infusion (0.75 microg/h) plus the selective aldosterone blocker eplerenone (100 mg. kg(-1). day(-1)). Aldosterone induced severe hypertension at 4 wk [systolic blood pressure (SBP), 210 +/- 3 mmHg vs. vehicle, 131 +/- 2 mmHg, P < 0.001], which was partially attenuated by eplerenone (SBP, 180 +/- 7 mmHg; P < 0.001 vs. aldosterone alone and vehicle). No significant increases in myocardial interstitial collagen fraction or hydroxyproline concentration were detected throughout the study. However, histopathological analysis of the heart revealed severe coronary inflammatory lesions, which were characterized by monocyte/macrophage infiltration and resulted in focal ischemic and necrotic changes. The histological evidence of coronary lesions was preceded by and associated with the elevation of cyclooxygenase-2 (up to approximately 4-fold), macrophage chemoattractant protein-1 (up to approximately 4-fold), and osteopontin (up to approximately 13-fold) mRNA expression. Eplerenone attenuated proinflammatory molecule expression in the rat heart and subsequent vascular and myocardial damage. Thus aldosterone and salt treatment in uninephrectomized rats led to severe hypertension and the development of a vascular inflammatory phenotype in the heart, which may represent one mechanism by which aldosterone contributes to myocardial disease.     

Reversal of elevated cardiac expression of TGFbeta1 and endothelin-1 in OLETF diabetic rats by long-acting calcium antagonist

The effects of calcium channel blockers (CCBs) on complications associated with diabetes mellitus (DM) have been well studied in clinical and basic science investigations. Cardiovascular complications are a common feature of type 2 DM, and insulin resistance is an early clinical manifestation of type 2 DM. CCBs are widely used to treat cardiovascular diseases in patients with DM. In this study, we used a spontaneous type 2 diabetic rat model, Otsuka Long-Evans Tokushima Fatty (OLETF) rats, at a highly insulin-resistant stage with modest hyperglycemia. We examined cardiac expression of transforming growth factor-beta(1) (TGFbeta(1)) and endothelin-1 (ET-1) in male OLETF rats. At 8 weeks of age, OLETF rats were treated for 12 weeks with the long-acting CCB benidipine (1 mg/kg/day or 3 mg/kg/day, po, n = 12), with hydralazine hydrochloride (3 mg/kg/day, po, n = 12), or with vehicle (OLETF, n = 12), and male age-matched genetic control Long-Evans Tokushima Otsuka (LETO, n = 12) rats were used. Blood pressure was significantly higher in OLETF rats than in LETO rats, and benidipine treatment at both dosages in OLETF rats for 12 weeks did not significantly reduce blood pressure, whereas hydralazine treatment significantly lowered blood pressure in OLETF rats. Hydralazine and both dosages of benidipine significantly reduced upregulated cardiac ET-1 levels in OLETF rats. Plasma and cardiac TGFbeta1 levels were remarkably higher in OLETF rats compared with LETO rats and were normalized by treatment with benidipine (3 mg/kg/day). Our results suggest that CCBs are effective in normalizing upregulated cardiac TGFbeta1 and ET-1 levels at the insulin-resistant stage in OLETF rats, which may improve cardiac morphology and function in this rat model without altering blood pressure and plasma glucose levels. In contrast, hydralazine treatment also normalizes cardiac ET-1 levels while significantly reducing blood pressure.     

Eplerenone inhibits the intracrine and extracellular actions of angiotensin II on the inward calcium current in the failing heart. On the presence of an intracrine renin angiotensin aldosterone system

The influence of chronic administration of eplerenone on the intracrine as well as on the extracellular action of angiotensin II (Ang II) on L-type inward calcium current was investigated in the failing heart of cardiomyopathic hamsters (TO-2).For this, eplerenone (200 mg/kg/day) was administered orally to 2 month-old cardiomyopathic hamsters for a period of 3 months. Measurements of the peak inward calcium current (I(Ca)) was performed in single cells under voltage clamp using the whole cell configuration. The results indicated that eplerenone suppressed the intracrine action of Ang II (10(-)(8) M) on peak I(Ca) density. Moreover, the intracellular dialysis of the peptide did not change the time course of I(Ca) inactivation in animals treated chronically with eplerenone. The extracellular administration of Ang II (10(-)(8) M) incremented the peak I(Ca) density by only 20+/-8% (n=30) compared with 38+/-4% (n=35) (P<0.05) obtained in age-matched cardiomyopathic hamsters not exposed to eplerenone. Interestingly, the inhibitory of eplerenone (10(-7) M) on the intracrine action of Ang II was also found, in vitro, but required an incubation period of, at least, 24 h. The inhibitory action of eplerenone on the intracellular action of Ang II was partially reversed by exposing the eplerenone-treated cells to aldosterone (10 nM) for a period of 24 h what supports the view that: a) the mineralocorticoid receptor(MR) was involved in the modulation of the intracrine action of the peptide; b) the effect of eplerenone on the intracrine as well as on the extracellular action of Ang II was related ,in part, to a decreased expression of membrane-bound and intracellular AT1 receptors. In conclusion: a) eplerenone inhibits the intracrine action of Ang II on inward calcium current and reduces drastically the effect of extracellular Ang II on I(Ca); b) aldosterone is able to revert the effect of eplerenone; c) the mineralocorticoid receptor is an essential component of the intracrine renin angiotensin aldosterone system.     

人参皂甙Rd抑制大鼠局灶性脑缺血后趋化因子CXCL1和γ-干扰素的蛋白表达

目的：研究人参皂甙Rd（Ginsenoside-Rd,GS-Rd）在大鼠局灶性脑缺血后对炎症趋化因子CXCL1和γ-干扰素（Interferon-γ,IFN-γ）的影响。方法：将SD大鼠随机分为5组：正常组（n=5）,假手术组（n=5）,GS-Rd对照组（n=5）,大脑中动脉栓塞模型（MCAO）组（n=20）,MCAO＋GS-Rd组（n=20）。正常组不做任何处理;假手术组进行大脑中动脉栓塞手术,但不插入栓线;GS-Rd对照组给予腹腔注射10 mg/Kg GS-Rd,不进行手术;MCAO组（n=20）和MCAO＋GS-Rd组（n=20）进行大脑中动脉栓塞手术,术后2小时拔出栓线,MCAO＋GS-Rd组在术前15分钟腹腔注射10 mg/Kg GS-Rd。在12小时、1天、3天、7天四个时间点分别提取脑组织蛋白,通过液相芯片技术检测CXCL1,IFN-γ含量。结果：正常组,假手术组和GS-Rd对照组组间CXCL1,IFN-γ含量无统计学差异;与三个对照组相比,MCAO组和MCAO＋GS-Rd组中CXCL1,IFN-γ蛋白含量均有明显增加（P〈0.05）;而与MCAO组相比,MCAO＋GS-Rd组CXCL1,IFN-γ的生成明显减少（P〈0.05）。结论：10 mg/Kg GS-Rd预处理可有效抑制大鼠短暂性脑缺血后CXCL1,IFN-γ的生成;通过抑制炎症反应,GS-Rd可能在神经保护中发挥重要的作用。     

罗格列酮对肺纤维化大鼠肺动脉壁结缔组织生长因子表达的影响

目的:观察过氧化物酶体增殖活化受体γ(PPAR-γ)激动剂罗格列酮(RSG)对肺纤维化大鼠肺动脉壁结缔组织生长因子(CTGF)上调、Ⅰ型和Ⅲ型胶原沉积的影响。方法:48只雄性SD大鼠,随机分为以下4组:博莱霉素(BLM)+生理盐水(NS)组(n=21)、BLM+RSG组(n=9)、NS+NS组(n=9)和NS+RSG组(n=9)。气管内一次性滴注BLM(5mg/kgbw),RSG灌胃(3mg/(kg.d),14d)。整体实验,气管滴注后第14天观察;离体实验,气管滴注BLM后第14天,分离大鼠的肺动脉,并用RSG培养液和单纯培养液孵育(37℃,5%CO2,24h)。结果:在整体水平,与对照大鼠相比,BLM模型大鼠肺动脉壁的CTGF免疫阳性表达增强,CTGF蛋白含量、Ⅰ型和Ⅲ型胶原含量、Ⅰ/Ⅲ胶原比值均增高(均P0.05);RSG能阻止上述指标的异常变化(均P0.05);在离体水平,RSG能阻止BLM模型大鼠肺动脉壁CTGF的上调(P0.05),但对Ⅰ型和Ⅲ型胶原沉积无明显影响(P0.05)。结论:RSG能直接作用于肺动脉壁,阻止肺纤维化大鼠肺动脉壁CTGF的上调,这可能是其减轻动脉壁结构重塑的机制之一。     

Angiotensin II AT1 receptors regulate ACE2 and angiotensin-(1-7) expression in the aorta of spontaneously hypertensive rats

When increased in vascular tissues, angiotensin-converting enzyme 2 (ACE2), a carboxypeptidase that hydrolyzes angiotensin II to angiotensin-(1-7), may augment the growth inhibitory and vasodilatory effects of the heptapeptide. We investigated the regulation of ACE2 and angiotensin-(1-7) expression in aortas and carotid arteries of 12-wk-old male spontaneously hypertensive rats (SHR) by determining the effect of sustained angiotensin type 1 (AT(1)) receptor blockade with olmesartan (10 mg.kg(-1).day(-1), n = 13) compared with those that received atenolol (30 mg.kg(-1).day(-1), n = 13), hydralazine (10 mg.kg(-1).day(-1), n = 13), or vehicle (n = 21). Systolic blood pressures were approximately 30% lower (P < 0.05) in rats treated for 2 wk with olmesartan compared with vehicle-treated rats. Both atenolol and hydralazine produced similar decreases in systolic blood pressure. ACE2 mRNA in the thoracic aorta of olmesartan-treated rats (n = 8) was fivefold greater (P < 0.05) than that in vehicle-treated rats (n = 16), whereas atenolol (n = 8) or hydralazine (n = 8) had no effect. Immunostaining intensities in rats treated with olmesartan (n = 5) were also associated with increased (P < 0.05) ACE2 and angiotensin-(1-7) in thoracic aorta media compared with vehicle-treated rats. In contrast, immunostaining intensities for both ACE2 and angiotensin-(1-7) were not different from vehicle (n = 5) in carotid arteries of SHR medicated with either atenolol (n = 5) or hydralazine (n = 5). A comparison of vessel wall dimensions showed that olmesartan selectively reduced the thoracic aorta media-to-lumen ratio (P < 0.05) and media thickness (P < 0.05) without an effect on carotid artery morphometry. Compared with vehicle-treated SHR, vascular hypertrophy determined from media and lumen measurements was not changed in SHR given either atenolol or hydralazine. These data represent the first report of ACE2 and angiotensin-(1-7) expression in the aorta and carotid arteries of SHR. Increased ACE2 and angiotensin-(1-7) in association with altered dimensions of the thoracic aorta but not carotid arteries in response to olmesartan treatment provides evidence that this pathway is regulated by AT(1) receptors and may be important in mediating the pressure-independent vascular remodeling effects of angiotensin peptides.     

靶向C—Raf-1基因的反义核酸抗乙型肝炎病毒活性研究

目的：通过体内外实验验证靶向c-Raf-1基因的反义核酸是否具有抑制乙型肝炎病毒（HBV）的活性。方法：设计靶向c-Raf-1基因的反义核酸,并在细胞水平进行体外抗HBV活性筛选,通过RT-PCR检测c-Raf-1基因mRNA水平的变化,通过体内药效学实验进一步验证反义核酸的抗HBV效果。结果：经体外筛选,靶向c-Raf-1基因的反义核酸Raf-3145具有相对明显的抑制HBV表面抗原（HBsAg）的作用,并可剂量依赖性地抑制c-Raf-1基因的表达;体内药效学结果显示,反义核酸Raf-3145在30 mg/kg剂量下对HBsAg的表达具有一定的抑制作用。结论：经体内外活性评价,初步确定了靶向宿主基因c-Raf-1的反义核酸具有一定的抑制HBsAg表达的活性,也进一步验证了c-Raf-1基因可以作为抗HBV药物设计的候选靶点。     

海藻色素糖蛋白对肝癌细胞Caspase-3和Bax蛋白表达的影响

目的：研究麒麟菜海藻色素糖蛋白（SPG）对肝癌细胞Caspase-3和Bax蛋白表达的影响。方法：将50只皮下接种H22肝癌细胞株的小鼠随机分为5组,每组10只。高、中、低剂量组分别每天经口灌胃给予100、50、10mg/kg的SPG,肿瘤对照组灌胃生理盐水,连续10d。环磷酰胺组隔天腹腔注射环磷酰胺20mg/kg.bw。取肝癌组织用MTT法测定各组肝癌细胞增殖活性,免疫组化法检测各组肝癌组织Caspase-3和Bax蛋白表达水平。结果：高剂量组Caspase-3和Bax蛋白表达率分别为40.20%和38.10%,而肿瘤对照组分别为5.00%和4.68%,差异均有显著性（P〈0.05）。高剂量SPG组和肿瘤对照组的肝癌细胞增殖活性分别为0.711±0.028和1.135±0.032,差别有显著性（P〈0.05）。结论：SPG可促进肝癌细胞Caspase-3和Bax蛋白表达,诱发肝癌细胞凋亡。     

蝮蛇毒介导小鼠急性心肌损伤及心肌组织TNF-α表达的研究

目的探讨蝮蛇毒介导小鼠心肌组织中TNF-α蛋白表达变化与不同蝮蛇毒剂量、中毒时间的关系。方法确定LD50蝮蛇毒注射剂量。将96只小鼠随机分为空白对照组、蛇毒低剂量(0.5mg/kg)组、蛇毒高剂量(1.0mg/kg)组,并按中毒时间设4水平组,即3、6、24、48h组,采集小鼠心肌标本,免疫组化检测心肌组织中TNF-α蛋白的表达情况。结果蛇毒低剂量组及蛇毒高剂量组小鼠心肌TNF-α蛋白表达明显高于空白对照组(均P0.05);而蛇毒高剂量组明显高于低剂量组(P0.05);6h组明显高于3h组(P0.05);24h组高于6h组(P0.05);48h组高于24h组(P0.05)。随着蝮蛇毒剂量的增加、中毒时间的延长,心肌TNF-α蛋白的表达越高。结论蝮蛇毒导致小鼠急性心肌损伤,TNF-α蛋白表达增高,且随着蝮蛇毒剂量的增加、中毒时间的延长,心肌TNF-α蛋白的表达越高,呈一定的时间、剂量效应趋势,提示TNF-α蛋白过度表达可能是蝮蛇毒导致小鼠急性心肌损伤的机制之一。