转化生长因子-β蛋白及mRNA基因在人类系膜增生性肾小球肾炎中的表达

目的：探讨转化生长因子－β（TGF－β）蛋白及mRNA基因在系膜增生性肾小球肾炎中的表达及其在发病中的作用。方法：采用常规病理,免疫组织化学及原位杂交方法对人系膜增生性肾小球肾炎组织进行染色,并经医学图像分析系统进行分析,结果：在正常对照组,TGF－β蛋白及TGF－β1mRNA小肾小管上皮细胞呈极弱表达,肾小球内TGF－β蛋白及mRNA未见阳性表达,在系膜增生肾小球肾炎组织中,TGF－β蛋白在肾近曲小管上皮细胞胞浆内呈强阳性表达,肾小球球囊壁及系膜区呈阳性,肾小球与肾小管阳性表达与正常对照组相比均具有显性差异（P＜0．01）,TGF－β1mRNA阳性表达位于肾近曲小管上皮细胞胞浆内和肾小球系膜区,肾小球和肾小管上皮细胞TGF－β1mRNA阳性表达与正常对照组相比差异也有显性（P＜0．01）。结论：系膜增生性肾小球炎时TGF－β蛋白及TGF－β1mRNARNA阳性表达与正常对照组相比差异也有显性（P＜0．01）。结论：系膜增生性肾小球肾炎时TGF－β蛋白及TGF－β1mRNA在肾小球与肾小管表达均显增高,进一步显示TGF－β在肾小球系膜细胞增生及肾小管间质纤维化中所起的重要作用。     

细胞粘附分子在IgA肾病患者肾组织中的表达──定量免疫组织化学研究

本研究应用免疫组织化学方法及计算机图象分析系统,对４７例ＩｇＡ肾病患者肾组织中细胞间粘附分子－１（ＩＣＡＭ－１）和血管细胞粘附分子－１（ＶＣＡＭ－１）的变化进行了定量研究。以此进一步探讨细胞粘附分子在ＩｇＡ肾病肾脏损伤中的作用变化机理。本文的结果显示：ＩＣＡＭ－１和ＶＣＡＭ－１在ＩｇＡ肾病肾小球内表达均增加,且与肾小球病变程度有密切关系。肾小球病变越重,上述粘附分子在肾小球内表达越强。重度ＩｇＡ肾病患者肾小球内ＩＣＡＭ－１和ＶＣＡＭ－１的表达最强。在伴有炎症细胞浸润的肾间质中ＩＣＡＭ－１也有存在。这一结果说明,ＩＣＡＭ－１和ＶＣＡＭ－１表达强度的变化与ＩｇＡ肾病的发展有密切的关系。     

原位杂交检测组织金属蛋白酶抑制因子—1 mRNA在IgA肾病肾脏中的表达

为探讨组织蛋白酶抑制因子在IgA肾病肾小球硬化中的作用。本文用原位杂交法研究组织金属蛋白酶抑制因子－1（TIMP－1）mRNA在3种类型IgA肾病（轻度系膜增生型、中度系膜增生型及局灶型）肾组织中的表达。结果表明：TIMP－1 mRNA原位杂匀的阳性信号主要分布于部分肾小管上皮细胞内。在肾小球系膜细胞也可检测到TIMP－1 mRNA阳性信号,但较肾小管弱,中度系膜增生型的TIMP－1 mRNA阳笥信号明显地较其他两型IgA肾病为强。以上结果表明：由肾小管上皮细胞及系膜细胞合成的TIMP－1通过抑制基质金属酶的活性,可导致细胞外基质（ECM）在肾小管间质及肾小球内过度沉积,从而对肾小球硬化有促进作用。     

碱性成纤维细胞生长因子对大鼠肾小球系膜细胞原癌基因c—fos和c—…

在建立大鼠肾小球系膜细胞体外培养方法的基础上,通过＾３Ｈ－ＴｄＲ参入实验,ＲＮＡ印迹分析和斑点杂交观察ｂＦＧＦ对ＭＣ ＤＮＡ合成及原癌基因ｃ－ｆｏｓ和ｃ－ｍｙｃ表达的影响。     

慢性肾小球肾炎患者血清HGF、Cys-C、TAFI水平变化及其临床诊断价值

目的:分析慢性肾小球肾炎患者血清肝细胞生长因子(HGF)、胱抑素C(Cys-C)、凝血酶激活的纤溶抑制物(TAFI)水平的变化及其临床诊断价值。方法:选择我院2017年1月～2018年5月收治的71例慢性肾小球肾炎患者作为慢性肾小球肾炎组及同期于本院进行健康体检的83例作为健康对照组。检测进而比较两组血清HGF、Cys-C、TAFI水平,分析以上指标和患者肾功能的相关性及对慢性肾小球肾炎的诊断价值。结果:慢性肾小球肾炎组血清HGF、Cys-C、TAFI水平均显著高于对照组(P0.05)。慢性肾小球肾炎患者治疗后血清HGF、Cys-C、TAFI水平均显著低于治疗前(P0.05)。慢性肾小球肾炎患者血清HGF、Cys-C、TAFI水平和肾功能指标(肌酐(Scr)、尿素氮(BUN)、尿酸(UA))均呈显著正相关(P均0.05)。血清HGF水平诊断慢性肾小球肾炎的曲线下面积为0.826,敏感度和特异度分别为0.747和0.746;血清Cys-C水平诊断慢性肾小球肾炎的曲线下面积为0.821,敏感度和特异度分别为0.687和0.859;血清TAFI水平诊断慢性肾小球肾炎的曲线下面积为0.816,敏感度和特异度分别为0.855和0.647;血清HGF、Cys-C、TAFI水平联合检测诊断慢性肾小球肾炎的曲线下面积为0.951,敏感度和特异度分别为0.831和0.757。结论:慢性肾小球肾炎患者血清HGF、Cys-C及TAFI水平均明显升高,联合检测血清HGF、Cys-C及TAFI可能作为慢性肾小球肾炎诊断及预评估参考指标。     

逆转录PCR及原位杂交检测人IgA肾病肾脏中TGF-β mRNA

为研究转化生长因子－β（ＴＧＦ－β）在ＩｇＡ肾病病理机制中的作用,本文用逆转录ＰＣＲ（ＲＴ－ＰＣＲ）法及原位杂交法检测ＩｇＡ肾病患者肾活检组织中的ＴＧＦ－βｍＲＮＡ。结果证实：在人正常肾和ＩｇＡ肾病肾组织中均可检测到ＴＧＦ－βｍＲＮＡ的ＲＴ－ＰＣＲ扩增产物;ＴＧＦ－βｍＲＮＡ原位杂交的阳性信号主要分布于肾小管和集合管上皮细胞内,肾小球内的阳性信号较弱,且强度与系膜的增生程度有关。     

剪切应力对毛细血管内皮细胞代谢的影响

建立的平行平板流协腔装置适用于研究血管内皮细胞代谢对剪切流场的响应。将培养的人胚肾小球血管单层内皮细胞置于剪应力分别为５＊１０＾－５Ｎ／ｃｍ＾２,１＊１０＾－４Ｎ／ｃｍ＾２和１．５＊１０＾－４Ｎ／ｃｍ＾２的定常层流中剪切２５小时。     

肾小球系膜细胞间隙连接的通讯功能及其影响因素

本文利用激光扫描共聚焦显微镜及荧光漂白后荧光回复技术对人肾小球系膜细胞间隙连接的通讯功能及其影响因素进行了观察。结果发现,体外培养的肾小球系膜细胞在融合生长状况下,相邻两细胞有间隙连接结构形成。不同浓度的血管紧张素Ⅱ对肾小球系膜细胞间隙连接的通讯功能具有促进作用,而甲基强的松龙则具有抑制作用。     

碱性成纤维细胞生长因子对大鼠肾小球系膜细胞原癌基因c-fos和c-myc表达的影响

在建立大鼠肾小球系膜细胞（ＭＣ）体外培养方法的基础上,通过３Ｈ－ＴｄＲ参入实验,ＲＮＡ印迹分析和斑点杂交观察ｂＦＧＦ对ＭＣＤＮＡ合成及原癌基因ｃ－ｆｏｓ和ｃ－ｍｙｃ表达的影响．结果表明,ｂＦＧＦ作用于ＭＣ１８ｈ,ＭＣ的３Ｈ－ＴｄＲ参入率明显增加（Ｐ＜０．０５）,２４ｈ达到高峰（Ｐ＜０．０１）;ｂＦＧＦ显著诱导原癌基因ｃ－ｆｏｓ和ｃ－ｍｙｃ表达,其表达活性分别于３０ｍｉｎ和１ｈ达到高峰．提示ｂＦＧＦ是ＭＣ的强效丝裂原,其对ＭＣＤＮＡ合成的促进作用与诱导原癌基因ｃ－ｆｏｓ和ｃ－ｍｙｃ表达有关．     

剪切应力对毛细血管内皮细胞代谢的影响

建立的平行平板流动腔装置适用于研究血管内皮细胞代谢对剪切流场的响应。将培养的人胚肾小球血管单层内皮细胞置于剪应力分别为５×１０－５Ｎ／ｃｍ２,１×１０－４Ｎ／ｃｍ２和１．５×１０－４Ｎ／ｃｍ２的定常层流中剪切２５小时,样品中的内皮素分泌量用放射免疫法测定。结果表明,剪应力水平对内皮细胞内皮素的代谢活动有显著影响。与静态培养对照,低水平的剪应力（５×１０－５Ｎ／ｃｍ２、１×１０－４Ｎ／ｃｍ２）促进内皮素的分泌,而较高水平的剪应力（１．５×１０－４Ｎ／ｃｍ２）抑制内皮素的分泌;剪应力对内皮素累积含量的影响比之分泌速率更大     

Histochemical studies on the mechanism of macromolecule leakage across the glomerular capillary wall. Barrier effect of the anionic groups of the capillary wall against the serum protein leakage

For the purpose of revealing the barrier effect of the anionic groups of glomerular capillary wall against the serum protein leakage, morphologic and histochemical observations were made on the rat kidney perfused in situ with three kinds of cationic macromolecules different in chemical characteristics followed by blood flow restoration. The polyethyleneimine perfusion resulted in the complete disapperance of ionized anionic groups of glomerular capillary and the massive protein leakage through glomeruli by blood flow restoration. Cationic ferric colloid perfusion induced moderate protein leakage, and avidin perfusion was less in neutralization effect of anionic groups and the protein leakage was of least. The protein leakage from glomeruli, however, was stopped or markedly suppressed soon after the blood flow restoration by the newly formed functioning anionic barrier probably by some particular serum protein deposition. The findings indicate that the deionization of the glomerular capillary wall will not be responsible for the persistent albuminuria.     

Prostaglandin synthesis in isolated glomeruli.

Prostaglandins are thought to play an important role in the local regulation of glomerular blood flow and in the release of renin from the juxtaglomerular apparatus. We therefore examined prostaglandin synthesis by isolated rat glomeruli. Isolated glomeruli were either prelabeled with [14C] arachidonic acid or were incubated with [14C] arachidonic acid for the entire experimental incubation in Krebs buffer. Prostaglandin synthesis was determined by thin layer radio-chromatography of acid extracts of the supernatant solutions. Indomethacin inhibitable synthesis of small amounts of 6-keto-PGF1 alpha, the metabolite of prostacyclin (PGI2,) and larger amounts of PGF2 alpha, and PGE2, and possibly thromboxane B2 (TXB2) by isolated glomeruli could be demonstrated with either prelabeling or direct incubation. These findings support the hypothesis that prostaglandins are produced within the glomerulus where they may affect local glomerular blood flow and function.     

Prostaglandin synthesis in isolated glomeruli

Prostaglandins are thought to play an important role in the local regulation of glomerular blood flow and in the release of renin from the juxtaglomerular apparatus. We therefore examined prostaglandin synthesis by isolated rat glomeruli. Isolated glomeruli were either prelabeled with [14_C] arachidonic acid or were incubated with [14_C] arachidonic acid for the entire experimental incubation in Krebs buffer. Prostaglandin synthesis was determined by thin layer radiochromatography of acid extracts of the supernatant solutions. Indomethacin inhibitable synthesis of small amounts of 6-keto-PGF_1α, the metabolite of prostacyclin(PGI₂,) and larger amounts of PGF_2α, and PGE₂, and possibly thromboxane B₂ (TXB₂) by isolated glomeruli could be demonstrated with either prelabeling or direct incubation. These findings support the hypothesis that prostaglandins are produced within the glomerulus where they may affect local glomerular blood flow and function.     

Quantitative relationship of renal blood flow with size and density of glomeruli in rat postnatal ontogenesis

The aim of the study was to find the quantitative relationship of postnatal changes in the glomeruli anatomic structure with the blood flow in kidneys. Kidney development was studied in 4-, 12-, 30-, and 65-day-old Wistar rats. Diameters of glomerulus (Dgl, μm), afferent and efferent arterioles (Daf and Def), and the glomeruli density (Ngl/mm⁻³) were measured posthumously. Volumes of one (Ugl, μm³) and all glomeruli (ΣUgl, mm³/cm³) and the glomeruli arterioles lumen (Saf, Sef, μm²) were calculated. The renal specific blood flow (SBF per unit of kidney weight, KW, mg) was measured by the laser-Doppler flowmeter (in perfusion units, p.u.) under sodium barbamyl narcosis. To assess the relationship of these parameters, we found their allometric association. We have found that, during postnatal growth, glomeruli morphological parameters vary according to the equations: Dgl = 7.1□KW^0.41, Ugl = 187□KW^1.23, Ngl = 5309□KW^−0.63 (KW, mg and for one kidney), Saf = 1.1□Ugl^0.35, and Sef = 6.3□Ugl^0.14. The renal SBF in 4-, 12-, and 65-day-old rats increases according to SBF = 6.7□(ΣUgl)^0.98. The renal SBF calculated per unit of glomeruli volume varies a little with age.     

Quantitative relationship of renal blood flow with size and density of glomeruli in rat postnatal ontogenesis

The aim of the study was to find the quantitative relationship of postnatal changes in the glomeruli anatomic structure with the blood flow in kidneys. Kidney development was studied in 4-, 12-, 30-, and 65-day-old Wistar rats. Diameters of glomerulus (Dgl, microm), afferent and efferent arterioles (Daf and Def), and the glomeruli density (Ngl, mm(-3)) were measured posthumously. Volumes of one ((see text of symbol))V gl, microm3) and all glomeruli (see text for symbol)(sigma(see text for symbol)Vgl, mm3/cm3) and the glomeruli arterioles lumen (Saf and Sef, microm2) were calculated. The renal specific blood flow (SBF per unit of kidney weight, KW) was measured by the laser-Doppler flowmeter (in perfusion units, p.u.) under sodium barbamyl narcosis. We have found that, during postnatal growth, glomeruli morphological parameters vary according to the equations: Dgl = 7.1 (see text for symbol) KW0.41, (see text for symbol)V gl = 187 (see text for symbol) KW1.23, Ngl = 5309 (see text for symbol) KW-0.63 (KW, mg and for one kidney), Saf = 1.1 (see text for symbol)V gl 0.35, and Sef = 6.3 (see text for symbol) V gl 0.14. The renal SBF in 4-, 12-, and 65-day-old rats increases according to SBF = 6.7 (see text for symbol) (sigma( see text for symbol)V gl)0.98. The renal SBF calculated per unit of glomeruli volume varies a little with age.     

In Utero Intra-cardiac Tomato-lectin Injections on Mouse Embryos to Gauge Renal Blood Flow

The formation and perfusion of developing renal blood vessels (apart from glomeruli) are greatly understudied. As vasculature develops via angiogenesis (which is the branching off of major vessels) and vasculogenesis (de novo vessel formation), perfusion mapping techniques such as resin casts, in vivo ultrasound imaging, and micro-dissection have been limited in demonstrating the intimate relationships between these two processes and developing renal structures within the embryo. Here, we describe the procedure of in utero intra-cardiac ultrasound-guided FITC-labeled tomato lectin microinjections on mouse embryos to gauge the ontogeny of renal perfusion. Tomato lectin (TL) was perfused throughout the embryo and kidneys harvested. Tissues were co-stained for various kidney structures including: nephron progenitors, nephron structures, ureteric epithelium, and vasculature. Starting at E13.5 large caliber vessels were perfused, however peripheral vessels remained unperfused. By E15.5 and E17.5, small peripheral vessels as well as glomeruli started to become perfused. This experimental technique is critical for studying the role of vasculature and blood flow during embryonic development.     

The glomerular apparatus and renal blood flow in rats during postnatal ontogenesis

In anesthetized Wistar rats of the age of 4, 12, 30 and 65 days, the renal blood flow (RBF) with laser-Doppler flowmetry, diameter, number and density of glomeruli, diameter of afferent and efferent arterioles were measured. We found that during rat development the RBF increased 2.8-fold, the diameter of glomeruli--3.6-fold, diameter of afferent arterioles--1.8-fold, and the efferent arterioles--rose 1.3-fold. The number of glomeruli increased 1.3-fold, but their density decreased 16-fold.     

Histochemical studies on the mechanism of macromolecule leakage across the glomerular capillary wall

Summary For the purpose of revealing the barrier effect of the anionic groups of glomerular capillary wall against the serum protein leakage, morphologic and histochemical observations were made on the rat kidney perfused in situ with three kinds of cationic macromolecules different in chemical characteristics followed by blood flow restoration. The polyethyleneimine perfusion resulted in the complete disappearance of ionized anionic groups of glomerular capillary and the massive protein leakage through glomeruli by blood flow restoration. Cationic ferric colloid perfusion induced moderate protein leakage, and avidin perfusion was less in neutralization effect of anionic groups and the protein leakage was of least. The protein leakage from glomeruli, however, was stopped or markedly suppressed soon after the blood flow restoration by the newly formed functioning anionc barrier probably by some particular serum protein deposition. The findings indicate that the deionization of the glomerular capillary wall will not be responsible for the persistent albuminuria.     

Is the renin-angiotensin system involved in urinary concentration mechanisms?

Renin release elicited by i.v. injection of loop-diuretics was used to study the effects of angiotensin II (AII) on intrarenal hemodynamics. The vasoconstrictive action of intrarenally synthesized AII predominates in the efferent glomerular arteriole. Such a vasoconstrictive effect could affect blood flow in the vasa recta which stem from efferent arterioles of juxtamedullary glomeruli. Renin secretion and renal inner medullary blood flow (tissue clearance of 133Xe) were simultaneously measured before and after frusemide-induced renin release. The relationship between renin secretion and renal inner medullary blood flow was inverse. Changes in renal medullary blood flow may be physiological determinants of medullary osmolality and renal concentration ability. The intrarenal role of AII in urinary concentration recovery after frusemide was examined. Inhibition of renin release by propranolol or AII-blockade (by saralasin or Hoe 409) delayed recovery of urinary osmolality. In the conscious rat, propranolol slowed down recovery of the cortico-papillary gradient for sodium. Its vasoconstrictive action on the efferent glomerular arteriole might enable the renin-angiotensin system to participate in the control of renal excretion of salt and water.     

Responsiveness of renal glomeruli to adenosine in streptozotocin-induced diabetic rats dependent on hyperglycaemia level.

Glomerular filtration rate (GFR) in response to adenosine precursor, NAD, and glomeruli contractility in response to adenosine were evaluated in streptozotocin-induced diabetic rats with severe (blood glucose 27.8 +/- 1.2 mmol/L) and moderate hyperglycaemia (18.2 +/- 0.9 mmol/L) compared with nondiabetic (ND)-rats. In anaesthetised rats, basal GFR was greater in moderately diabetic rats compared with severely diabetic rats (p < 0.05) and ND-rats (p < 0.02). Intravenous infusion of 5 nmol x min(-1) x kg(-1) NAD reduced GFR and renal plasma flow (RPF) in diabetic rats but had no effect on these parameters in ND-rats. Moreover, NAD-induced reduction of GFR and RPF was greater in rats with severe diabetes (41% and 30%, respectively) than in with moderate diabetes (25% and 26%, respectively). Theophylline (0.2 micromol x min(-1) x kg(-1) ) abolished renal response to NAD. Isolated glomeruli contraction in response to adenosine, assessed by glomerular 3H-inulin space reduction, was lowered in moderately diabetic-group and enhanced in severely diabetic-group. compared with ND-group (p < 0.05). Adenosine A1-receptor antagonist DPCPX inhibited adenosine-induced glomeruli contraction. This differential response of diabetic renal glomeruli to adenosine suggests that impaired glomerular contractility in response to adenosine could be responsible for hyperfiltration in moderate diabets, whereas, the increased adenosine-dependent contractility of glomeruli in severe diabetes may increase the risk of acute renal failure in this condition.