醛糖还原酶拮抗剂治疗大鼠血管再狭窄

背景醛糖还原酶增强氧化应激,促进细胞分裂增殖,激活转录因子、核因子кB(NF-кB),但在血管再狭窄中起的作用报告不多 .目的 研究醛糖还原酶(AR)在大鼠再狭窄血管中的表达,探讨AR表达与内膜增生的关系,AR拮抗剂依帕司他对再狭窄血管中AR的表达及AR的抑制作用.方法 健康雄性SD大鼠24只,随机分成对照组(A组)、手术未干预组(B组)及手术 依帕司他组(C组),每组8只.用通气-干燥法损伤B组及C组大鼠右侧颈总动脉,左侧颈总动脉作为对照.分别于7 d及14 d后处死大鼠,HE染色以观察内膜及中膜增生情况,并计算内膜、中膜面积及其比值,FISH原位杂交及免疫组化检测AR及NF-кB的表达.结果 1)术后第7天损伤侧血管内膜增生明显,第14天后内膜增生进一步加重,对照血管无明显增生.2)依帕司他明显抑制血管内皮损伤后血管内膜增生、血管内膜面积、血管内膜与中膜面积比值明显低于未干预组(P＜0.05或P＜0.01).3)AR及NF-кB 在损伤侧血管中的表达明显强于对侧.依帕司他明显抑制AR及NF-кB 的表达量(P＜0.05或P＜0.01).结论 依帕司他对血管内膜的增生及再狭窄的形成有抑制作用,伴有AR及NF-кB 的表达受抑.     

阿托伐他汀对大鼠再狭窄血管醛糖还原酶表达及内膜增生的抑制作用

目的研究醛糖还原酶在大鼠再狭窄血管中的表达,探讨醛糖还原酶表达与内膜增生之间的关系,以及阿托伐他汀对醛糖还原酶表达及内膜增生的抑制作用。方法24只健康雄性SD大鼠,随机分成手术未干预组、假手术组及阿托伐他汀组,每组8只。用通气—干燥法损伤手术未干预组及阿托伐他汀组中的右侧颈总动脉,左侧颈总动脉作为对照。分别于第7天及14天处死动物,HE染色以观察内膜及中膜增生情况,并计算内膜和中膜面积及其比值,FISH原位杂交及免疫组织化学检测醛糖还原酶及核因子κB的表达。结果术后第7天损伤侧血管内膜增生明显,第14天内膜增生进一步加重,对照血管无明显增生。阿托伐他汀组血管内膜面积、内膜与中膜面积比值明显低于手术未干预组(P<0.05或P<0.01)。醛糖还原酶及核因子κB在损伤侧血管中的表达明显强于对照侧。阿托伐他汀组醛糖还原酶及核因子κB的表达明显低于手术未干预组(P<0.05或P<0.01)。结论醛糖还原酶可能参与了大鼠颈动脉损伤后内膜增生及再狭窄的形成。阿托伐他汀可抑制血管内膜增生及再狭窄的形成。阿托伐他汀可能是通过抑制醛糖还原酶及核因子κB而抑制再狭窄的形成。     

依帕司他对醛糖还原酶和核转录因子-κB表达及血管平滑肌细胞增殖的影响

目的研究依帕司他对醛糖还原酶(AR)、核转录因子(NF)-κB表达及血管平滑肌细胞(VSMC)增殖的影响,探讨依帕司他抗细胞增殖的可能机制。方法取大鼠胸主动脉段行VSMC培养,用高浓度葡萄糖(22.5mmol/L)诱导AR基因表达,然后加入不同浓度的依帕司他,培养3d后行VSMC计数,并用免疫组化、RT-PCR及原位杂交的方法检测NF-κB及AR的表达。结果①随依帕司他浓度的增加,VSMC计数逐渐减少。②高浓度葡萄糖时NF-κB表达强阳性,加入不同浓度的依帕司他后,NF-κB表达逐渐减弱。③高浓度葡萄糖可明显诱导AR基因表达,依帕司他对其表达有抑制作用,且呈剂量依赖性。结论①高浓度葡萄糖可促进VSMC增殖,依帕司他可抑制这一增殖作用,且具有剂量依赖性。②依帕司他可抑制AR及NF-κB的表达。③依帕司他可能是通过抑制AR及NF-κB的表达继而抑制VSMC的增殖。     

黄芩苷对糖尿病大鼠组织醛糖还原酶活性及肾脏细胞凋亡的影响

目的研究醛糖还原酶(AR)、凋亡相关蛋白Bcl-2和Bax与糖尿病肾病(DN)的关系,探讨醛糖还原酶抑制剂(ARIs)对DN的治疗作用。方法雄性Wistar大鼠40只,随机分为正常对照组、糖尿病组、依帕司他组、黄芩苷组,每组10只。除正常对照组外,其余各组大鼠腹腔注射STZ(60mg/kg),72h后建立糖尿病大鼠模型。依帕司他组灌胃依帕司他10mg.kg-1.d-1,黄芩苷组灌胃黄芩苷150mg.kg-1.d-1。16w后处死大鼠,测定晶体、肾脏组织AR活性,观察Bcl-2、Bax的表达,取部分肾皮质病理观察、电镜观察细胞凋亡的形态学变化。结果与正常对照组相比,糖尿病组晶体、肾脏皮质AR活性明显升高(P<0.001),两治疗组AR活性明显较糖尿病组降低(P<0.01),而血糖无明显变化。糖尿病组肾脏Bcl-2、Bax蛋白表达增加,治疗组的Bcl-2蛋白表达较糖尿病组增多,而Bax蛋白表达较糖尿病组减少。透射电镜下见糖尿病组肾脏肾小管上皮细胞呈典型的凋亡形态学改变,治疗组大鼠肾组织细胞凋亡改变明显减轻。糖尿病组肾小球基底膜增厚,系膜区域扩大,治疗组病变减轻。结论ARIs通过抑制AR活性,调节Bax、Bcl-2的表达抑制细胞凋亡,延缓DN的发展。黄芩苷对AR抑制作用与依帕司他相似。     

血管内放射治疗对血管成形术后再狭窄及核因子-кB活性的影响

目的:观察32P液体球囊血管近距离照射预防血管成形术后再狭窄的量效关系及其对核转录因子核因子-кB (NF-кB)活性的影响,以探讨其防治再狭窄的可能作用机制.方法:24只大耳白兔,建立兔双侧髂动脉粥样硬化狭窄模型,随机选择一侧髂动脉血管成形术并分别给予9.1Gy组、21.8Gy组和33.4Gy 组32P液体球囊血管照射治疗(每组n=8),另一侧髂动脉灌注造影剂,作为自身对照(对照组,n=24).术后5周行血管造影并取材进行光镜观察、NF-кB、胰岛素样生长因子-1(IGF-1)免疫组织化学染色,用计算机图像分析测量新生内膜面积、中膜面积、管腔面积及免疫组化染色阳性面积百分比.结果:①光镜观察: 9.1Gy组:与对照组比病变无明显差异;21.8Gy组:内弹力膜完整无明显断裂;管腔轻度狭窄,新生内膜面积明显减小,内膜层泡沫样细胞及脂质沉积均不明显;中膜平滑肌呈轻度增厚, 排列轻度紊乱;33.4Gy组:内弹力膜破坏,管腔明显狭窄,内膜可见大量泡沫细胞及脂质沉积,大量炎性细胞浸润及细胞外基质不定形物质沉积,中膜平滑肌明显萎缩变薄,其中4例血管腔内可见血栓形成.②免疫组化染色:9.1Gy组:NF-кB、IGF-1蛋白表达与对照组无明显差异(P>0.05);21.8Gy组:NF-кB、IGF-1蛋白表达量中等,呈灶状散在分布于内皮细胞、平滑肌细胞及泡沫细胞,与对照组相比有明显差异(P<0.01);③33.4Gy组:NF-кB、IGF-1蛋白少量表达,呈散在性分布,明显低于9.1Gy组和21.8Gy组(P<0.01).结论:32P液体球囊在一定的吸收剂量范围内确可安全有效地防止血管成形术后再狭窄形成,其机制可能为抑制NF-кB及其靶基因的活化,从而抑制血管平滑肌细胞的增殖、迁移.     

罗格列酮对大鼠血管再狭窄的作用及其发生机制

目的探讨罗格列酮在体内对斯普拉-道来(Sprague-Dawley,SD)大鼠血管再狭窄的影响及其发生机制。方法 40只SD大鼠随机分为2组:治疗组喂罗格列酮[(5mg·kg-1·d-1)溶于饮水中]4周;对照组喂等量的0.9%氯化钠溶液4周。4周后建立大鼠颈总动脉再狭窄模型,继续上述喂养2周后取材,免疫组织化学法检测核因子-кB(nuclearfactor-кB,NF-кB),TUNEL法检测细胞凋亡率;用形态测量法检测血管内膜厚度和面积。结果术后2周,与对照组比较,治疗组内膜厚度变薄,内膜面积减少,差异有统计学意义[(12.451±4.593)μmvs.(32.455±9.092)μm,P0.05;(0.017±0.005)mm2vs.(0.052±0.013)mm2,P0.05]。与对照组比较,治疗组NF-кB的水平明显受抑制,细胞凋亡率显著升高,差异有统计学意义(0.0814±0.0021vs.0.1015±0.0038,P0.01;32.58%±1.75%vs.24.36%±2.46%,P0.01)。结论罗格列酮可阻止大鼠血管平滑肌细胞增殖及内膜增生。     

醛糖还原酶抑制剂依帕司他改善大鼠糖尿病膀胱病变组织的功能

目的 研究醛糖还原酶(AR)、一氧化氮(NO)与糖尿病膀胱病(DCP)的关系,探讨醛糖还原酶抑制剂(ARIs)依帕司他对DCP的治疗作用. 方法 雄性SD大鼠60只分三组;正常组、糖尿病组和治疗组.测定膀胱组织AR活性,NO含量及一氧化氮合酶(NOS)活性. 结果与正常组相比,糖尿病组膀胱组织内AR活性、NO含量和NOS活性升高(P<0.01),治疗组AR活性、NO含量和NOS活性均低于糖尿病组(P<0.05). 结论 依帕司他通过抑制AR活性,调节NO含量变化及NOS活性,减轻氧化应激反应,改善膀胱组织的功能.     

川芎嗪对糖尿病鼠醛糖还原酶活性及视网膜细胞凋亡影响

雄性Wistar大鼠40只,随机平均分为4组,正常对照组、糖尿病组、糖尿病依帕司他(10mg/kg/d)治疗组、糖尿病川芎嗪(100mg/kg/d)治疗组,腹腔注射链脲佐菌素(65mg/kg)诱发糖尿病.16周后,处死大鼠,分离晶体,测定组织AR活性,观察视网膜Bcl-2、Bax的表达.结果1.与正常对照组相比,糖尿病组AR活性明显升高(P＜0.001),依帕司他治疗组、川芎嗪治疗组AR活性较糖尿病组明显降低(P＜0.01),而血糖无明显变化.2.糖尿病组视网膜Bcl-2、Bax蛋白表达明显增加,依帕司他治疗组、川芎嗪治疗组的Bcl-2、Bax蛋白表达较糖尿病组明显减少.结论1.AR过度激活通过促进Bcl-2、Bax蛋白的表达诱导细胞凋亡,而参与DR的发生与发展.2.ARts通过抑制醛糖还原酶活性,调节Bax、Bcl-2的表达抑制细胞凋亡,延缓DR的发展.3.川芎嗪对AR抑制作用与典型ARIs依帕司他相似,且价格低、副作用少,值得临床推荐应用.     

阿托伐他汀对高葡萄糖诱导的血管平滑肌细胞增殖的抑制作用

目的研究阿托伐他汀对醛糖还原酶(AR)和核因子NF-κB的表达及血管平滑肌细胞增殖的影响,探讨阿托伐他汀抗细胞增殖的可能机制.方法用高浓度葡萄糖诱导大鼠血管平滑肌细胞(VSMC)AR基因表达,然后加入不同浓度的阿托伐他汀,培养3 d后用台盼蓝染色行细胞计数.并采用RT-PCR、免疫组化、原位杂交等方法,观察阿托伐他汀对NF-κB和AR基因表达及VSMC增殖的影响.结果 1)随着阿托伐他汀浓度的提高,VSMC计数逐渐减少.2)正常浓度葡萄糖(5.6 mmol/L)时, NF-κB表达不明显,高浓度葡萄糖(22.5 mmol/L)时,NF-κB表达强阳性,阿托伐他汀则可明显抑制这一表达,0.1 μmol/L阿托伐他汀时, NF-κB表达即有降低,10 μmol/L阿托伐他汀几乎完全抑制NF-κB的表达.3)高浓度葡萄糖可明显诱导AR基因的表达,阿托伐他汀对其表达有抑制作用,且呈剂量依赖性.与高浓度葡萄糖组相比,阿托伐他汀0.1、1、10 μmol/L分别使VSMC的AR mRNA下调12%、45%和80%(P均＜0.05).结论 1)阿托伐他汀可抑制VSMC增殖.2)阿托伐他汀可抑制AR及NF-κB的表达.3)阿托伐他汀可能是通过抑制AR及NF-κB的表达继而抑制VSMC的增殖.     

高脂血症对内皮剥脱术后血管炎症反应的影响

目的探讨高脂血症对内皮剥脱术后血管炎症反应的影响。方法建立大鼠主动脉内皮剥脱后内膜增生模型,利用免疫组化及Western blot方法检测血管壁NF-κB和iNOS的表达变化,并结合形态学分析研究高脂血症大鼠内皮剥脱后血管炎症反应及新生内膜增生情况。结果主动脉内皮剥脱后内膜呈弥漫性增厚,管腔变小,增厚的内膜以VSMC为主,中膜VSMC排列紊乱。高脂血症明显促进了增生的内膜中NF-κB和iNOS的表达。结论高脂血症通过促进NF-κB表达进而诱导iNOS的表达,加剧了内皮剥脱后的血管炎症反应。     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.     

N-acetyl-L-cysteine combined with mesalamine in the treatment of ulcerative colitis： Randomized,placebo-controlled pilot study

AIM： To evaluate the effectiveness and safety of oral N-acetyl-L-cysteine （NAC） co-administration with mesalamine in ulcerative colitis （UC） patients.
METHODS： Thirty seven patients with mild to moderate UC were randomized to receive a four-wk course of oral mesalamine （2.4 g/d） plus N-acetyl-L-cysteine （0.8 g/d） （group A） or mesalamine plus placebo （group B）. Patients were monitored using the Modified Truelove-Witts Severity Index （MTWSI）. The primary endpoint was clinical remission （MTWSI ≤ 2） at 4 wk. Secondary endpoints were clinical response （defined as a reduction from baseline in the MTWSI of ≥ 2 points） and drug safety. The serum TNF-α, interleukin-6, interleukin-8 and MCP-1 were evaluated at baseline and at 4 wk of treatment. RESULTS： Analysis per-protocol criteria showed clinical remission rates of 63% and 50% after 4 wk treatment with mesalamine plus N-acetyl-L-cysteine （group A） and mesalamine plus placebo （group B） respectively （OR = 1.71; 95% CI： 0.46 to 6.36; P = 0.19; NNT = 7.7）. Analysis of variance （ANOVA） of data indicated a significant reduction of MTWSI in group A （P = 0.046） with respect to basal condition without significant changes in the group B （P = 0.735） during treatment. Clinical responses were 66% （group A） vs 44% （group B） after 4 wk of treatment （OR = 2.5; 95% CI： 0.64 to 9.65; P = 0.11; NNT = 4.5）. Clinical improvement in group A correlated with a decrease of IL-8 and MCP-1. Rates of adverse events did not differ significantly between both groups.
CONCLUSION： In group A （oral NAC combined with mesalamine） contrarily to group B （mesalamine alone）, the clinical improvement correlates with a decrease of chemokines such as MCP-1 and IL-8. NAC addition not produced any side effects.     

Delorme's transrectal excision for internal rectal prolapse

Surgical therapy of functional outlet obstruction in patients with internal rectal intussusception may include abdominal, perineal, or transrectal procedures. Because abdominal procedures often result in significant physiologic impact but unrelieved constipation, the authors have elected Delorme's transrectal excision for management of these patients. Since a short-term placebo effect attends many therapies, this report describes results of transrectal excision only after a threeyear postoperative period. Delorme's transrectal excision of internal intussusception accomplished sustained symptomatic relief in over 70 percent of otherwise refractory constipated patients. The association of internal intussusception with other abnormalities underscores the importance of defining both anatomic and functional components when selecting patients whose constipation may require surgical therapy. Critical technical elements, surgical pitfalls, and potential complications of the procedure are discussed.Poster presentation at the meeting of The American Society of Colon and Rectal Surgeons, Toronto, Canada, June 11 to 16, 1989.     

The oral glucose tolerance test: A comparison of the time points on the basis of limit values,normal dispersion,and reproducibility

Summary Time points in the glucose tolerance test (GTT) are compared on the basis of limit values, dispersion within a reference population, and reproducibility. We suggest using the distance between a limit value and the median reference value as a measure of the magnitude of abnormality. The distance between 140 mg/100 ml and the median fasting plasma glucose value is chosen as a standard distance and limits for other points in the GTT are calculated to equal this standard distance of abnormality. We suggest that the probability of correctly interpreting an inividual result is directly related to the reproducibility of the test and inversely related to the percentage of the total range of values which is dispersed among the normal population. The ratio of reproducibility to percentage normal dispersion is proposed as an index of the probability of correctly interpreting an individual result. According to this index, the probability of correct interpretation varies in order: fasting plasma glucose concentration>3-h>2-h>0.5-h>1-h plasma glucose concentration.