TGF-β1/p38MAPK通路在介导单侧输尿管梗阻大鼠肾间质纤维化过程中的作用

目的 探讨TGF-β1/p38MAPK信号传导通路在老年单侧输尿管梗阻(UUO)大鼠肾间质纤维化过程中的作用.方法 将老年SD大鼠随机分为假手术组、UUO模型组、SB203580治疗组.分别至UUO术后第3、7、14天处死,取梗阻侧肾脏,观察病理改变;采用免疫组织化学方法检测各组大鼠肾小管上皮细胞中转化生长因子(TGF)-β1、p-p38、α-平滑肌肌动蛋白(SMA)的表达情况.结果 与假手术组比较,模型组及治疗组TGF-β1、p-p38MAPK 、α-SMA的表达显著升高(P＜0.05);而与模型组相比较,治疗组TGF-β1、p-p38MAPK、α-SMA的表达显著降低(P＜0 05).TGF-β1、p-p38MAPK和α-SMA三者的表达呈正相关(TGF-β1与p38MAPK之间r1=0.745,P＜0.05;p-p38MAPK与α-SMA之间r2=0.659,P＜0.05;TGF-β1与α-SMA之间r3=0.770,P＜0.05).结论 TGF-β1/ p38MAPK通路的激活可上调肾小管上皮细胞α-SMA的表达,促进上皮细胞转分化,并可能是调控UUO大鼠肾间质纤维化主要信号通路之一.     

去甲斑蝥素抑制单侧输尿管梗阻大鼠肾小管上皮-间充质转分化的实验研究

目的:利用单侧输尿管梗阻(UUO)大鼠模型,观察去甲斑蝥素(norcantharidin,NCTD)对肾小管上皮细胞间质转分化的影响。方法:SD大鼠随机分为假手术组(Sham组)、模型组(UUO组)和UUO+NCTD治疗组(NCTD组),每组6只。采用单侧(左)输尿管结扎方法,建立梗阻性肾小管间质纤维化大鼠模型,Sham组只游离输尿管而不结扎。术后14d处死动物,取结扎侧肾脏,同时处死Sham组大鼠,取出肾组织。采用免疫组化、Western Blot及RT-PCR检测α平滑肌肌动蛋白(α-SMA)、E钙黏蛋白(E-cadherin)与转化生长因子β1(TGF-β1)蛋白的mRNA表达的变化。结果:免疫组化、Western Blot及RT-PCR均显示,与Sham组比较,UUO组肾组织α-SMA和TGF-β1的mRNA表达升高、E-cadherin蛋白和mRNA表达降低(P<0.05);NCTD能抑制UUO大鼠肾组织α-SMA和TGF-β1表达的上调,上调E-cadherin的表达(P<0.05)。结论:NCTD具有抑制UUO大鼠肾小管上皮-间充质转化的作用,该作用可能与抑制TGF-β1的过表达有关。     

红花对肾间质纤维化实验大鼠肾小管上皮细胞表型转化的抑制作用

目的 目的观察红花对结扎单侧输尿管诱导的肾间质纤维化实验大鼠肾小管上皮细胞表型转化的抑制作用.方法 结扎单侧输尿管方式复制肾间质纤维化实验动物模型,分别给以红花4.0 g*kg-1*d-1、缬沙坦10 mg*kg-1*d-1经口灌服,采用免疫组化、原位杂交方法检测大鼠肾脏肾小管上皮细胞表型转化标志物α-平滑肌肌动蛋白(α-SMA)及Ⅲ型胶原的表达.结果 单侧输尿管结扎大鼠肾脏α-SMA动蛋白和Ⅲ型胶原的阳性面积及积分光密度较假手术组明显增强,红花及缬沙坦可显著性抑制其表达.结论 红花可以通过抑制肾小管上皮细胞的表型转化拮抗肾间质纤维化.     

肾组织核心蛋白聚糖和转化生长因子-β1的表达与肾间质纤维化的关系

目的探讨转化生长因子β 1(TGF-β1)及核心蛋白聚糖(DCN)表达与肾间质纤维化之间的关系,及α- 平滑肌肌动蛋白(α-SMA)和单核巨噬细胞浸润在肾脏疾病中的变化. 方法收集30例不同肾脏病肾活检患者的临床和病理资料,应用免疫组织化学染色方法,检测TGF-β1及DCN在肾间质内的表达,以及α-SMA和CD 68的表达.并分析它们的表达与肾间质纤维化之间的关系. 结果肾间质病变严重程度与血清肌酐、肾内TGF-β1和DCN表达程度成正相关,与内生肌酐清除率成负相关.肾间质浸润的巨噬细胞主要见于间质大量炎性细胞浸润的部位及纤维化间质和病变肾小球周围;α-SMA表达可见于血管壁、纤维化间质、增厚的肾小球囊壁或病变肾小球周围以及变性萎缩的肾小管管周,它们的表达与肾间质纤维化程度呈正相关. 结论随着肾间质纤维化程度的加重,DCN和 TGF-β1的表达逐渐增加,同时伴有间质单核巨噬细胞的浸润和间质细胞的激活,参与肾间质纤维化的进程.     

吗替麦考酚酯对大鼠肾小管上皮-间充质转化的作用及其机制探讨

目的 探讨吗替麦考酚酯(MMF)对腺嘌呤致慢性肾衰竭大鼠肾间质纤维化及肾小管上皮-间充质转化(EMT)的作用,以及该作用同肾组织血管内皮生长因子(VEGF)和分化抑制因子(Id2、Id3)变化的关系.方法 雄性Wistar大鼠64只,分对照组(16只)、慢性肾衰竭组(24只,腺嘌呤125 mg·kg-1·d-1灌胃)、MMF干预组(24只,腺嘌呤125 mg·kg-1·d-1+MMF 15 mg·kg-1·d-1灌胃).分别于第2、4、6、8周处死动物,收集血、尿标本测血肌酐、尿蛋白.取肾组织,Masson染色作肾间质纤维化程度评分,免疫组化法检测肾组织α-平滑肌肌动蛋白(α-SMA)、转化生长因子β1(TGFβ1)表达,RT-PCR测肾组织TGFβ1、VEGF mRNA表达,Western blot测肾组织α-SMA、VEGF及Id2、Id3蛋白表达.结果 (1)慢性肾衰竭组、MMF干预组血肌酐均显著高于对照组,但慢性肾衰竭组、MMF干预组之间无显著差异;MMF干预组第6、8周时24 h尿蛋白显著低于慢性肾衰竭组.(2)慢性肾衰竭组、MMF十预组在第6、8周肾间质纤维化程度显著重于对照组,但MMF干预组间质纤维化程度显著低于慢性肾衰竭组.(3)慢性肾衰竭组、MMF干预组在第6、8周肾组织(α-SMA、TGFβ1表达较对照组明显增强,但MMF干预组肾组织α-SMA表达在第6、8周明显低于慢性肾衰竭组,TGFβ1表达在第2、4、6周亦显著低于慢性肾衰竭组.(4)慢性肾衰竭组第8周肾组织VEGF蛋白表达低于对照组,MMF干预组肾组织VEGF表达第6、8周显著高于慢性肾衰竭组.(5)MMF干预组肾组织Id2、Id3蛋白表达高于慢性肾衰竭组,在第4、6周差异显著.结论 MMF具有减轻慢性肾衰竭大鼠肾间质纤维化、抑制肾小管上皮细胞EMT的作用,该作用可能与MMF下调肾组织TGFβ1表达及上调VEGF、Id2、Id3表达有关,其确切机制需进一步探讨.     

缬沙坦和螺内酯抑制自发性高血压大鼠心肌纤维化的对比研究

目的比较缬沙坦和螺内酯对自发性高血压大鼠(SHR)心肌纤维化的抑制作用及二者对SHR心肌整合素β1和纤维黏连蛋白(FN)表达的影响,探讨SHR心肌纤维化的机制。方法将18只6周龄SHR随机均分成3组缬沙坦组(缬沙坦30mg·kg-1·d-1),螺内酯组(螺内酯20mg·kg-1·d-1)和SHR组,并与雄性同周龄WKY6组只比较,实验期14周。比较血压、左室重量/体重(LVM/BW)、胶原容积分数(CVF)和血管周围胶原面积(PVCA),并用免疫组化法检测4组大鼠心肌整合素β1和FN的表达。结果(1)缬沙坦组和螺内酯组血压〔(1125±88)mmHg和(1316±51)mmHg〕、LVM/BW〔(284±014)×10-3和(322±015)×10-3〕、CVF〔(321±022)%和(400±028)%〕、PVCA〔(062±015)%和(094±056)%〕均显著低于SHR组(P<005),缬沙坦组与螺内酯组比较,差异有显著性(P<005);(2)SHR组心肌整合素β1和FN的表达明显高于WKY组〔其积分吸光度值(A)分别为(15687±2033)、(5430±800)和(45665±4127)、(15718±2830),P<001,而缬沙坦组和螺内酯组FN的沉积较SHR组(45665±4127)均显著减少〔分别为(20047±1680)和(21358±800),P<005〕;缬沙坦组整合素β1的表达(9892±135)较SHR组下降(P<005),而螺内酯组整合素β1的表达与SHR组差异无显著性(P>005)。结论缬沙坦和螺内酯不仅可以良好控     

表没食子儿茶素没食子酸酯对肾间质纤维化大鼠的保护作用

目的 探讨表没食子儿茶素没食子酸酯[(-)-epigallocatechin-3-gallate,EGCG]对单侧输尿管梗阻(UUO)大鼠肾间质纤维化的保护作用.方法 将30只健康、雄性SD大鼠随机分为3组:分别为UUO组、假手术组(SOR)、和UUO+EGCG组,术前分别给UUO组和SOR组生理盐水灌胃,UUO+EGCG组EGCG 20 mg·kg-1·d-1灌胃,连续3 d.术后第8天分别处死各组大鼠用免疫组织化学方法测定转化生长因子β(TGF-β) 和P53的表达情况.行HE和Masson染色,动态观察肾间质形态学改变.结果 UUO+EGCG组P53的表达明显低于UUO组;UUO+EGCG组TGF-β的表达量(1.92±0.35)与UUO组(3.27±0.56)比较有明显的下降(P<0.01);能减轻胶原在肾间质的沉积,改善了肾脏病理改变.结论 EGCG对UUO所致的大鼠肾脏纤维化的形成有明显的抑制作用,其作用可能跟降低TGF-β的表达阻止细胞损伤有关.     

别嘌醇对2型糖尿病小鼠肾小管间质细胞表型改变的保护作用与机制

目的探讨别嘌醇对2型糖尿病db/db小鼠肾小管间质转分化的抑制作用及其可能机制。方法将db/m小鼠作为正常对照组(db/m组),将2型糖尿病db/db小鼠随机分为db/db小鼠对照组(db/db组)和db/db小鼠别嘌醇50 mg·kg-1·d-1干预组(db/db+A组),共干预12 w。疗程结束后检测小鼠血糖、血尿酸、血尿素氮、血肌酐、血甘油三酯和24 h尿蛋白水平;采用PAS及Masson染色观察肾脏组织病理学变化;免疫组化染色观察α-平滑肌肌动蛋白(α-SMA)、E-钙黏素(E-cadherin)、转化生长因子β(TGF-β)、抗增殖因子(prohibitin)的表达分布;Western印迹检测肾皮质TGF-β、prohibitin的蛋白表达水平。结果与正常对照组比较,db/db组血糖、血尿酸、血尿素氮、血肌酐、血甘油三酯和24 h尿蛋白水平均显著增高(P<0.01);肾组织TGF-β和α-SMA蛋白表达水平明显上调(P<0.01);而prohibitin、E-cadherin蛋白表达明显下降(P<0.01)。db/db+A组血尿酸、血尿素氮、24 h尿蛋白水平均较db/db组明显降低(P<0.01)。肾组织TGF-β和α-SMA蛋白表达水平也较db/db组有所下降;prohibitin、E-cadherin蛋白表达有所升高(P<0.01)。肾组织病理检查显示:db/db组肾小管上皮细胞空泡样变、肾小管管腔扩张、肾间质纤维增生等肾小管间质损伤明显,db/db+A组上述病变程度较db/db组减轻。结论别嘌醇对2型糖尿病小鼠肾小管间质损伤具有保护作用,其机制可能与恢复肾组织prohibitin含量、抑制TGF-β过度表达、进而抑制肾小管间质细胞转分化有关。     

过氧化物酶体增殖物激活受体γ激动剂对TGF-β1诱导肾成纤维细胞细胞外基质表达的影响

目的:研究过氧化物酶体增殖物激活受体γ(PPARγ)激动剂对转化生长因子β1(TGF-β1)致肾间质纤维化作用的影响,探讨其抗肾间质纤维化的潜在作用.方法:体外培养大鼠肾成纤维细胞株(NRK/49F),利用逆转录-聚合酶链反应(RT-PCR)观察PPARγ配体15d-PGJ2及其激动剂曲格列酮和齐格列酮对TGF-β1诱导的α-平滑肌肌动蛋白(α-SMA)、结缔组织生长因子(CTGF)、纤维连结蛋白(FN)和Ⅲ型胶原(ColⅢ)mRNA表达的影响,利用Western Blot方法观察PPARγ激动剂对TGF-β1诱导的FN蛋白表达的影响.结果:TGF-β1能显著增加NRK/49F细胞CTGF、FN和ColⅢmRNA表达,并呈剂量依赖和时间依赖效应.与TGF-β1刺激组相比,10 μM 15d-PGJ2、曲格列酮和齐格列酮预处理组α-SMA、CTGF、FN和ColⅢmRNA表达量显著减少,FN蛋白表达量显著下降.结论:PPARγ激动剂可抑制TGF-β1诱导的肾间质成纤维细胞CTGF表达和细胞外基质(ECM)合成.     

α-平滑肌肌动蛋白及转化生长因子-β1在高尿酸血症大鼠肾小管间质纤维化中的表达

目的观察α-平滑肌肌动蛋白(α-SMA)及转化生长因子-β1(TGF-β1)在高尿酸血症(HUA)大鼠肾小管间质纤维化(RIF)过程中的表达及意义。方法 SD大鼠36只,随机分为对照组(Con)、模型组及非布司他组,每组12只。氧嗪酸钾(OP)溶于0.5%的羧甲基纤维素钠(CMC)溶液,模型组和非布司他组用OP灌胃,1次/d,制备HUA模型,非布司他组另予非布司他灌胃,Con组仅予等量0.5%(CMC灌胃。于0、4、8周后检测血尿酸(SUA)、血肌酐(Scr)及BUN,收集24 h尿量,检测24 h尿蛋白定量。第0、4、8周末每组随机处死4只大鼠,HE染色及Masson染色观察肾组织病理变化,免疫组织化学法检测α-SMA、TGF-β1的表达。结果实验4周后,与Con组比较,模型组SUA、Scr、BUN升高(P0.05或P0.01),TGF-β1、α-SMA表达量增多(P0.01),肾小管间质发生纤维化。实验8周后,RIF加重;非布司他组RIF较模型组减轻(P0.01)。结论 HUA诱导肾组织TGF-β1、α-SMA表达量增加;α-SMA及TGF-β1表达增多促进RIF发生。     

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.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

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.     

Microbiology of human immunodeficiency virus anorectal disease

PURPOSE: Individuals who are seropositive for the human immunodeficiency virus are at high risk for opportunistic infection and anorectal disorders. Little prospective information is available regarding anorectal pathogens in these patients. METHODS: One hundred sixty-three HIV-seropositive patients presented to the colorectal clinic between 1989 and 1992. Forty-seven (29 percent) patients were thought to have an infectious process and were prospectively studied using a standardized multiculture protocol. RESULTS: Mean age was 33 (range, 19–59) years. All were male; high-risk behavior accounted for 87 percent of HIV transmissions. Presenting complaints included anorectal pain (79 percent), pus per anum (28 percent), and blood per anum (26 percent). Examination revealed perianal tenderness (60 percent), condyloma (38 percent), perianal ulcers (38 percent), and anal fissures (34 percent). Sixty-six sets of cultures were performed; 28 patients had one set, 15 had two sets, and 4 had three sets. Thirty-two of these 47 patients (68 percent) had positive cultures including herpes (50 percent), cytomegalovirus (25 percent),Neisseria gonorrhoeae (16 percent), chlamydia (16 percent), acidfast bacilli (2 percent), and others (9 percent). Six of 32 patients with positive cultures had more than one organism cultured. Sixteen (50 percent) patients with positive cultures were treated medically, 8 (25 percent) were treated surgically and 8 (25 percent) were treated with both modalities. Sixty-one procedures were performed on 17 patients for condylomata. Eighteen patients had 20 procedures for abscesses, 50 percent of whom had positive cultures for other than common bowel flora; all improved. Fourteen patients underwent 33 procedures for perianal fistulas.Mycobacterium fortuitum was cultured from one patient who required 13 procedures for abscesses and fistulas. Forty-five (96 percent) patients were followed for an average of 12.5 months ±2.9 SEM (range, 1–94 months). Symptoms were improved or resolved in 22 of 32 (69 percent) patients with positive cultures and in 11 of 13 (84 percent) with negative cultures. CONCLUSIONS: Specific pathogens may often be identified in human immunodeficiency virus-seropositive patients with anorectal disorders if aggressively sought. Although patients without specific pathogens identified may be expected to improve with planned empiric treatment, positive identification allows more directed therapy.