Suppression of experimental glomerulonephritis by antiserum against transforming growth factor beta 1

Glomerulonephritis is an inflammation of the kidney characterized by the accumulation of extracellular matrix within the damaged glomeruli, impaired filtration and proteinuria. In its progressive form, the disease destroys kidney function leading to uraemia and death, unless dialysis therapy or kidney transplantation is available. The pathogenesis of glomerulonephritis is incompletely understood, but the eliciting factor is thought often to be an immunological injury to mesangial and/or other resident cells in the glomeruli. We have used an animal model of acute mesangial proliferative glomerulonephritis to show that this disease is associated with increased production and activity of transforming growth factor beta 1 (TGF-beta 1), an inducer of extracellular matrix production. Here we report that administration of anti-TGF-beta 1 at the time of induction of the glomerular disease suppresses the increased production of extracellular matrix and dramatically attenuates histological manifestations of the disease. These results provide direct evidence for a causal role of TGF-beta 1 in the pathogenesis of the experimental disease and suggest a new approach to the therapy of glomerulonephritis.     

Expression of human proteoglycan in Chinese hamster ovary cells inhibits cell proliferation

In studying the functional role of an extracellular matrix proteoglycan, decorin, we have made observations that suggest a role for this proteoglycan in the control of cell proliferation. Extracellular matrices are made up of different combinations of collagens, elastin, hyaluronic acid, proteoglycans and various glycoproteins such as fibronectin. Most of these components can interact with cells, and much of the control of cell adhesion, migration and differentiation appears to be mediated by these interactions. Earlier studies have also attributed growth-regulatory activities to intact extracellular matrices, but the individual molecules responsible for these effects have not been characterized. We report here that Chinese hamster ovary (CHO) cell lines expressing human decorin from a stably transfected complementary DNA construct form a more orderly monolayer and grow to a lower saturation density than control cells lacking decorin. The extent of the morphological changes correlates with the level of decorin expression, and the saturation density is inversely proportional to it. The reduction in the saturation densities of the cell lines with the highest expression of decorin is more than 50%. These results reveal a novel growth inhibitory mechanism which may be related to contact inhibition of cell proliferation.     

鳖肾脏的基本病理组织学初步观察

鳖肾脏的基本病理组织变化以及肾小球肾炎为主,不同的病种和病程表现有异,大体上可分为增生性肾小球肾炎、渗出性肾小球肾炎、充血性肾小球肾炎和硬化性肾小球肾炎等。     

Amyloid beta-peptide is produced by cultured cells during normal metabolism.

Alzheimer's disease is characterized by the extracellular deposition in the brain and its blood vessels of insoluble aggregates of the amyloid beta-peptide (A beta), a fragment, of about 40 amino acids in length, of the integral membrane protein beta-amyloid precursor protein (beta-APP). The mechanism of extracellular accumulation of A beta in brain is unknown and no simple in vitro or in vivo model systems that produce extracellular A beta have been described. We report here the unexpected identification of the 4K (M(r) 4,000) A beta and a truncated form of A beta (approximately 3K) in media from cultures of primary cells and untransfected and beta-APP-transfected cell lines grown under normal conditions. These peptides were immunoprecipitated readily from culture medium by A beta-specific antibodies and their identities confirmed by sequencing. The concept that pathological processes are responsible for the production of A beta must not be reassessed in light of the observation that A beta is produced in soluble form in vitro and in vivo during normal cellular metabolism. Further, these findings provide the basis for using simple cell culture systems to identify drugs that block the formation or release of A beta, the primary protein constituent of the senile plaques of Alzheimer's disease.     

Negative regulation of transforming growth factor-beta by the proteoglycan decorin

Decorin is a small chondroitin-dermatan sulphate proteoglycan consisting of a core protein and a single glycosaminoglycan chain. Eighty per cent of the core protein consists of 10 repeats of a leucin-rich sequence of 24 amino acids. Similar repeats have been found in two other proteoglycans, biglycan and fibromodulin, and in several other proteins including Drosophila morphogenetic proteins. Expression of high levels of decorin in Chinese hamster ovary cells has a dramatic effect on their morphology and growth properties. We now report that this effect is due at least in part to the ability of decorin to bind transforming growth factor-beta, an autocrine factor that stimulates the growth of Chinese hamster ovary cells. As transforming growth factor-beta induces synthesis of decorin in many cell types, our results suggest that decorin may be a component of a feedback system regulating cell growth.     

Connective tissue growth factor induces transformation of renal fibroblasts into myofibroblasts

Connective tissue growth factor (CTGF) is thought to be a specific mediator of TGF-β profibrogenic effect, but the role of CTGF in the transformation of renal fibroblasts to myofibroblasts, which are the most important host cells in the renal chronic fibrosis, is still unknown. Aimed at observing the biological effect of CTGF, we set up a cell line overexpressing CTGF, and analyzed the amount of myofibroblast and the level of extracellular matrix Collagen Ⅲ mRNA in cultured cells. Our results show that CTGF can directly induce myofibroblasts formation, and increase the level of Collagen Ⅲ mRNA. This suggests that CTGF may be a novel pharmacotherapeutical target protein for the interference with renal fibrosis, thus providing useful theoretic possibility for clinical prevention of chronic progression of kidney disease.     

Atrial natriuretic peptide causes pre-glomerular vasodilatation and post-glomerular vasoconstriction in rat kidney

Atrial natriuretic peptide (ANP) can be extracted from rat hearts, and is found to increase fluid excretion by the kidneys when injected into test animals. The mechanism of ANP action is still unclear. ANP may reduce sodium reabsorption in the renal tubules, but it is also known that it increases the rate of glomerular filtration in the kidney, and relaxes preparations of smooth muscle, including one made from arteries that supply the kidney. To clarify its mode of action, we have studied directly the effects of semi-purified and synthetic ANP on blood vessels in the kidney of anaesthetized rats. We found that ANP causes a vasodilatation of the blood vessels which supply the glomeruli and a vasoconstriction of the arterioles which drain them. This substantiates the finding that increased filtration pressure participates in the natriuretic response.     

高脂血症大鼠肾脏Angptl4变化及辛伐他汀的保护作用

为探讨高脂血症大鼠（Rattus norregicus）肾脏中Angptl4的变化及辛伐他汀的保护作用,将45只雄性SD大鼠随机平均分为3组：正常对照组（NG）给予普通饲料喂养;高脂饮食组（HG）给予高脂饲料喂养;辛伐他汀组（SG）在高脂饲料喂养的基础上给予辛伐他汀10mg·kg^-1·d^-1水溶液灌胃。分别于第4、6、lO周时各组随机选取5只大鼠,检测大鼠24h尿蛋白及血清总胆固醇（TC）和低密度脂蛋白胆固醇（LDL-C）水平,石蜡切片HE染色观察大鼠肾小球的损害,免疫组化及Western blot检测大鼠肾脏Angptl4蛋白分布及表达。结果显示,于第4周开始,HG组和sG组大鼠血清TC和LDL-C水平升高。实验第10周时,HG组和SG组大鼠24h尿蛋白定量升高;大鼠肾组织HE染色光镜下观察显示：肾小球呈轻微分叶状,系膜区增宽和基质增多不明显,但肾小球内有较明显的空泡变性,SG组大鼠的病变明显轻于HG组;免疫组化和免疫印迹检查发现,大鼠肾脏Angptl4明显增多,SG组的表达较HG组有减少。研究认为,高脂血症可引起肾脏肾小球明显病变,可能是肾小球内的足细胞分泌Angptl4的增多引起;而辛伐他汀则可能使高脂饮食时足细胞分泌Angptl4减少,从而减轻高脂血症时肾小球受到的损害。     

700例慢性肾功能衰竭病因调查分析

通过分析慢性肾功能衰竭（chronic renal failure,CRF）病因的发病状况,证明早期及时治疗原发病对于降低或延缓CRF的重要性。对象和方法：分别从华西附一医院、华西附二医院、四川省医院、成都长城肾脏病医院选取确诊为CRF的700例患者进行面对面的问卷调查,得出相应的结果。调查结果显示：慢性肾功能衰竭在已明确的病因中仍以原发性肾小球肾炎为主要病因,其次为糖尿病肾病、高血压肾病、痛风性肾病。     

T-cell-expressed proprotein convertase furin is essential for maintenance of peripheral immune tolerance

Furin is one of seven proprotein convertase family members that promote proteolytic maturation of proproteins. It is induced in activated T cells and is reported to process a variety of substrates including the anti-inflammatory cytokine transforming growth factor (TGF)-beta1 (refs 2-4), but the non-redundant functions of furin versus other proprotein convertases in T cells are unclear. Here we show that conditional deletion of furin in T cells allowed for normal T-cell development but impaired the function of regulatory and effector T cells, which produced less TGF-beta1. Furin-deficient T regulatory (Treg) cells were less protective in a T-cell transfer colitis model and failed to induce Foxp3 in normal T cells. Additionally, furin-deficient effector cells were inherently over-active and were resistant to suppressive activity of wild-type Treg cells. Thus, our results indicate that furin is indispensable in maintaining peripheral tolerance, which is due, at least in part, to its non-redundant, essential function in regulating TGF-beta1 production. Targeting furin has emerged as a strategy in malignant and infectious disease. Our results suggest that inhibiting furin might activate immune responses, but may result in a breakdown in peripheral tolerance.     

松花粉对糖尿病肾病小鼠肾脏损伤的保护作用

 探讨松花粉对链脲霉素（streptozotocin,STZ）诱导糖尿病肾病小鼠肾脏损伤的保护作用及可能机制。成年雄性C57BL/6J（Wild Type）小鼠50 只,采用STZ 诱导糖尿病肾病小鼠动物模型。实验小鼠随机分为正常对照组（10 只）、STZ 模型组（40只）;STZ 模型组按150 mg/kg 腹腔注射STZ 一次,复制小鼠糖尿病肾病模型,对照组小鼠则腹腔注射柠檬酸钠缓冲液。在造模1周后,小鼠空腹血糖值大于11.1 mmol/L 视为糖尿病诱导成功,此后定期每周监测血糖和24 h 尿蛋白,持续出现蛋白尿的小鼠视为成模小鼠。成模糖尿病肾病小鼠28 只,将其随机分为STZ 模型组和松花粉治疗组各14 只;松花粉治疗组每天给予750 mg/kg体重的松花粉灌胃,正常对照组与STZ 模型组以等体积的0.9%氯化钠注射液灌胃。连续7 周后,检测小鼠血糖、24 h 尿微量白蛋白、尿肌酐、血肌酐、血浆尿素氮水平;计算肾重/体重比、肾脏组织丙二醛（MDA）含量等指标,并对肾脏组织切片行Masson 和Jones 六胺银染色;Western Blot 的方法观察p38 和ERK 信号分子的磷酸化激活情况。与模型组相比,松花粉治疗组肾重/体重比、肾小球细胞外基质增生程度和肾功能损害程度均明显减轻;松花粉能够降低糖尿病肾脏的MDA 含量并抑制p38 和ERK 信号分子的激活。由此可以看出,松花粉能通过抑制p38 和ERK 信号分子的磷酸化激活,减轻肾脏的氧化损伤和肾小球细胞外基质增生。     

几种齿鲸小肾的形态研究

本文对印度恒河豚、日本海江豚、东海江豚、真海豚和南方宽吻海豚的小肾和肾小球进行了测量和计数,并对小肾结构指数做了测定。结果表明,江豚、真海豚和南方宽吻海豚的相对肾重和小肾数明显大于恒河豚;真海豚和宽吻海豚的肾小球总数远大于江豚和恒河豚,恒河豚最少。但总的肾小球表面积、每克体重的肾小球表面积均低于相当体重的陆生哺乳动物。鲸类肾脏的小肾化虽然增加了肾小球的数量,但并没有增加肾的滤过面积。几种齿鲸的小肾结构指故无一定规律.也没有发现它与环境盐度的对应关系。因此,对于鲸类肾脏如何适应它们旺盛的生理代谢和小肾结构指数与肾浓缩能力的关系还需进一步研究。     

Activated T lymphocytes produce a matrix-degrading heparan sulphate endoglycosidase

We have previously found that lines of activated T lymphocytes specifically autosensitized to the basic protein of myelin (BP), on intravenous inoculation into syngeneic rats, were able to penetrate blood vessels, accumulate in the nervous system and cause experimental autoimmune encephalomyelitis (EAE). An important question is how effector T cells reach such targets outside the walls of blood vessels. To investigate this we have studied in vitro the interaction of anti-BP effector T lymphocytes with the basement membrane-like extracellular matrix produced by vascular endothelial cells. We now report that activated but not resting T lymphocytes produce an endoglycosidase capable of degrading heparan sulphate side chains of the proteoglycan scaffold of the extracellular matrix. Moreover, the anti-BP T lymphocytes respond to BP presented by extracellular matrix by markedly enhanced elaboration of the endoglycosidase. These results suggest that tissue-specific antigens on blood vessel walls could direct lymphocyte homing by activating enzymes that facilitate penetration of the subendothelial basal lamina. They also suggest that effector T lymphocytes can recognize antigen which is not associated with a major histocompatibility complex signal.     

Type IV collagens regulate BMP signalling in Drosophila

Dorsal-ventral patterning in vertebrate and invertebrate embryos is mediated by a conserved system of secreted proteins that establishes a bone morphogenetic protein (BMP) gradient. Although the Drosophila embryonic Decapentaplegic (Dpp) gradient has served as a model to understand how morphogen gradients are established, no role for the extracellular matrix has been previously described. Here we show that type IV collagen extracellular matrix proteins bind Dpp and regulate its signalling in both the Drosophila embryo and ovary. We provide evidence that the interaction between Dpp and type IV collagen augments Dpp signalling in the embryo by promoting gradient formation, yet it restricts the signalling range in the ovary through sequestration of the Dpp ligand. Together, these results identify a critical function of type IV collagens in modulating Dpp in the extracellular space during Drosophila development. On the basis of our findings that human type IV collagen binds BMP4, we predict that this role of type IV collagens will be conserved.     

Loss of integrin alpha(v)beta6-mediated TGF-beta activation causes Mmp12-dependent emphysema

Integrins are heterodimeric cell-surface proteins that regulate cell growth, migration and survival. We have shown previously that the epithelial-restricted integrin alpha(v)beta6 has another critical function; that is, it binds and activates latent transforming growth factor-beta (TGF-beta). Through a global analysis of pulmonary gene expression in the lungs of mice lacking this integrin (Itgb6 null mice) we have identified a marked induction of macrophage metalloelastase (Mmp12)--a metalloproteinase that preferentially degrades elastin and has been implicated in the chronic lung disease emphysema. Here we report that Itgb6-null mice develop age-related emphysema that is completely abrogated either by transgenic expression of versions of the beta6 integrin subunit that support TGF-beta activation, or by the loss of Mmp12. Furthermore, we show that the effects of Itgb6 deletion are overcome by simultaneous transgenic expression of active TGF-beta1. We have uncovered a pathway in which the loss of integrin-mediated activation of latent TGF-beta causes age-dependent pulmonary emphysema through alterations of macrophage Mmp12 expression. Furthermore, we show that a functional alteration in the TGF-beta activation pathway affects susceptibility to this disease.     

原发性肾病综合征合并急性肾衰竭43例临床病理分析

目的研究分析原发性肾病综合征合并急性肾衰竭的临床及病理特点,提高此类并发症的防治水平。方法自2004年6月至2006年6月在我科住院确诊的43例原发性肾病综合征（PNS）合并急性肾衰竭（ARF）病例（ARF组）和同期住院的肾功能正常的原发性肾病综合征43例（对照组）。对两组病人的临床表现及病理特点进行对比分析。结果ARF组病人血压（收缩压、舒张压）、血浆自蛋白浓度、血肌酐浓度、血尿素氮浓度与对照组比较有显著性差异（P〈0.05）。病理分析ARF组中、重度系膜增殖性肾小球肾炎发生率比对照组明显增高,（P〈0.01）;ARF组膜增生性肾小球肾炎多于对照组（P〈0.05）;肾间质改变较对照组显著增多（P〈0.01）。ARF组D-二聚体阳性率较对照组增多（P〈0.05）;血浆纤维蛋白定量也较对照组显著增高（P〈0.01）。结论PNS合并ARF病人肾小球病理改变较重多伴肾间质病变、高凝状态、血栓形成及血浆自蛋白浓度较低等。因此,对PNS合并ARF者宜尽早行肾穿刺活检术,以便明确其病理类型,根据病理特点选择有效的治疗方案,能大大改善病人的预后。     

Failure of familial Alzheimer's disease to segregate with the A4-amyloid gene in several European families

The gene coding for the amyloid protein, a component of neuritic plaques found in brain tissue from patients with Alzheimer's disease, has been localized to chromosome 21, and neighbouring polymorphic DNA markers segregate with Alzheimer's disease in several large families. These data, and the association of Alzheimer's disease with Down's syndrome, suggest that overproduction of the amyloid protein, or production of an abnormal variant of the protein, may be the underlying pathological change causing Alzheimer's disease. We have identified a restriction fragment length polymorphism of the A4-amyloid gene, and find recombinants in two Alzheimer's disease families between Alzheimer's disease and the A4-amyloid locus. This demonstrates that the gene for plaque core A4-amyloid cannot be the locus of a defect causing Alzheimer's disease in these families. These data indicate that alterations in the plaque core amyloid gene cannot explain the molecular pathology for all cases of Alzheimer's disease.     

鸡马立克氏病病毒与大肠杆菌混合感染症的病原分离鉴定

鸡呼吸道病流行病学调查的过程中,在韶关市某养鸡场观察到一群100日龄广西黄肉仔鸡发生了以呼吸道症状为主要临床表现的疾病,死亡率达10%.病理解剖学检查可见病鸡有单一或多器官肿瘤病变(4/8)、典型腹膜炎病变(3/8)和肿瘤与腹膜炎病变并存(2/8).在病鸡肿瘤组织切片中可见典型MD病理组织学变化.采用CEF接种方法,从有肿瘤和腹膜炎病鸡的羽髓液中分离到MDV.在病鸡的心、肝、脾等脏器分离到大肠杆菌O78血清型菌株.研究结果表明:HVT疫苗免疫鸡群MDV强毒株和E.coliO78血清型菌株以混合感染形式存在,病鸡出现以呼吸道症状为主的临床表现,死亡率明显增加.     

Role of plasminogen activator and plasminogen activator inhibitor type-1 in luteolysis—a minireview

This minireview summarized our recent studies on the role of plasminogen activator (PA) and inhibitor type-1 (PAI-1) in luteolysis. We have demonstrated that (1) both tissue type and urokinase type plasminogen activators (tPA and uPA) and a plasminogen activator inhibitor type-1 (PAI-1) were present in the corpus luteum of rat and rhesus monkey; (2) decrease in progesterone production in corpus luteum was well correlated with a sharp increase in tPA (but not uPA) and PAI-1 secretion; (3) exogenous tPA decreased luteal progesterone synthesis while monoclonal antibodies increased progesterone production; (4) interferony inhibited luteal progesterone synthesis and stimulated tPA production while LH plus prolactin increased progesterone production and decreased tPA (hut not uPA) activity in cultured luteal cells; (5) increase in proteolysis in the corpus luteum was also correlated with decrease in progesterone production in mouse. These data suggest that local degradation of extracellular matrix controlled by plasminogen activator and inhibitor is involved in the processes of luteolysis.     

Role of plasminogen activator and plasminogen activator inhibitor type-1 in luteolysis——a minireview

This minireview summarized our recent studies on the role of plasminogen activator (PA) and inhibitor type-1 (PAI-1) in luteolysis. We have demonstrated that (1) both tissue type and urokinase type plasminogen activators (tPA and uPA) and a plasminogen activator inhibitor type-1 (PAI-1) were present in the corpus luteum of rat and rhesus monkey; (2) decrease in progesterone production in corpus luteum was well correlated with a sharp increase in tPA (but not uPA) and PAI-1 secretion; (3) exogenous tPA decreased luteal progesterone synthesis while monoclonal antibodies increased progesterone production; (4) interferon y inhibited luteal progesterone synthesis and stimulated tPA production while LH plus pro-lactin increased progesterone production and decreased tPA (but not uPA) activity in cultured luteal cells; (5) increase in proteolysis in the corpus luteum was also correlated with decrease in progesterone production in mouse. These data suggest that local degradation of extracellular matrix controlled by plasminogen activator and inhibitor is involved in the processes of luteolysis.