Effect of autophagy on necroptosis of renal tubular epithelial cells in subtotal nephrectomy rats

AIM: To explore whether autophagy is involved in the excessive death of renal tubular epithelial cells in subtotal nephrectomy(SNx) rats and the relationship between autophagy and necroptosis in the kidney of SNx rats. METHODS: Male Sprague-Dawley rats were randomly assigned to control group(n=6) and SNx group(n=42). The rats in SNx group were subjected to SNx. Sham surgery was performed in the rats in control group. The rats in SNx group were divided into subgroups at 0, 4, 8 and 12 weeks(n=6) and the other rats in SNx group were divided into SNx+vehicle group, SNx+necrostatin-1(Nec-1) group and SNx+3-methyladenine(3-MA) group. The expression of RIP1, RIP3, LC3 and beclin-1 at mRNA and protein levels was measured at 0, 4, 8 and 12 weeks by qPCR and immunohistochemistry. The effects of Nec-1 or 3-MA on the protein expression of LC3-I, LC3-II and beclin-1, and production of reactive oxygen species(ROS) in the rat kidney were determined by Western blot and DCFH-DA staining. The death of renal tubular epithelial cells in the SNx rats was observed by TUNEL staining and electron microscopy. Finally, the effects of Nec-1 and 3-MA on blood urea nitrogen(BUN), serum creatinine(SCr) and the pathological changes of the renal tissues were analyzed. RESULTS: The highest mRNA and protein levels of RIP1, RIP3, LC3 and beclin-1 appeared at the 8th week after SNx(P<0.01). Compared with the rats in SNx+vehicle group, the protein over-expression of LC3-II/I and beclin-1, renal tubular epithelial cells with typical morphological features of necroptotic cell death and TUNEL-positive renal tubular cells were decreased in the SNx rats treated with Nec-1 and 3-MA(P<0.01), but 3-MA did not reduce the increased concentration of ROS. In addition, treatment with Nec-1 and 3-MA obviously reduced BUN, SCr(P<0.05), glomerulosclerosis index and tubulointerstitial injury score(P<0.01). CONCLUSION: Autophagy participates in the excessive death of renal tubular epithelial cells in SNx rats. Inhibition of autograph prevents necroptotic cell death of renal tubular cells, and alleviates chronic renal injury in SNx rats.     

Severe polyuria and polydipsia as major clinical signs in a horse with unilateral renal adenocarcinoma

A 14‐year‐old Swiss Warmblood gelding was presented with chronic severe polyuria, polydipsia and weight loss. At the time of admission, water intake was 240 l/day. On rectal examination, a large mass was identified in the left dorsal abdominal quadrant, which was shown to originate from the left kidney by transabdominal ultrasonographic examination. Unilateral nephrectomy via flank incision was performed under general anaesthesia. Histopathological examination of the tumour revealed a papillary renal adenocarcinoma. Successful outcome and survival was documented 13 months after surgery. Severe polyuria and polydipsia should be considered as major clinical signs for renal carcinoma in horses, which can be successfully treated with unilateral nephrectomy if no signs of metastatic spread are evident.     

六味地黄汤对5/6肾切除大鼠肾组织中细胞外基质的影响

目的观察六味地黄汤对慢性肾衰大鼠肾组织细胞外基质的影响,探讨其防治肾间质纤维化的可能机制。方法用5/6肾切除大鼠复制慢性肾衰模型,分空白对照组、假手术组、模型组和六味地黄汤组。术后8周,检测各组肾功能指标;HE染色观察肾脏组织病理变化,Masson染色检测肾脏中胶原纤维沉积;免疫组化法检测ColⅠ、ColⅢ、FN在肾组织中的表达;Western blot法检测PDGF-A、PDGF-B在肾组织中的蛋白表达水平。结果与假手术组比较,模型组24 h尿蛋白、血尿素氮和血肌酐明显升高(P<0.01);与模型组比较,六味地黄汤组24 h尿蛋白(P<0.01)、血尿素氮(P<0.01)和血肌酐(P<0.05)均明显降低。模型组HE和Masson染色显示大鼠肾间质纤维化明显,大量胶原纤维沉积,六味地黄汤组肾间质纤维化程度明显减轻,少量胶原纤维沉积。免疫组化法显示,与假手术组比较,模型组肾组织中ColⅠ、ColⅢ、FN蛋白水平均明显升高(P<0.01);与模型组比较,六味地黄汤组肾组织中ColⅠ、ColⅢ、FN蛋白水平均明显降低(P<0.05)。Western blot法显示,与假手术组比较,模型组肾组织中PDGF-A、PDGF-B蛋白水平均明显升高(P<0.01);与模型组比较,六味地黄汤组肾组织中PDGF-A、PDGF-B蛋白水平均明显降低(P<0.01)。结论六味地黄汤可能通过下调PDGF-A、PDGF-B表达,从而减少肾组织中细胞外基质的积聚而抑制肾间质纤维化。     

The use of computed tomography in the diagnosis of an ectopic ureter in a Quarter Horse filly

This report describes a 4‐month‐old Quarter Horse filly with an ectopic ureter. The filly presented with signs of urinary incontinence, which had been present since birth. Computed tomography (CT) examination and cystoscopy confirmed a diagnosis of a unilateral ectopic ureter. A nephrectomy of the left kidney was performed and renal function was closely monitored post operatively. The filly was treated for abdominal chylous effusion as a post operative complication. The filly survived to discharge from the hospital, and maintained normal urinary function at 12 months post operatively.     

Use of intraoperative impression smear cytology to guide successful treatment of a large renal cyst in a dog: a case report

A 13-yr-old Shih tzu was referred for surgical management of right-sided cranial abdominal mass, which corresponded to large, cavitated renal mass on ultrasonography, and was suspected to represent neoplasia. Intraoperative impression smear cytology (ISC) of the renal mass wall was consistent with benign renal cyst (RC), without evidence of neoplasia or infection. Deroofing and omentalisation were performed and histopathology was consistent with benign RC. Chronic kidney disease was diagnosed 4 mon postoperatively, however, the dog was asymptomatic, without cyst reoccurrence. Intraoperative ISC is an expedient and inexpensive diagnostic technique that can guide most appropriate treatment in dogs with large RCs.     

RENAL SONOGRAPHIC MEASUREMENTS IN THE DOG PRECEDING AND FOLLOWING UNILATERAL NEPHRECTOMY

The purpose of this study was to establish sonographic baseline values for normal kidneys initu and to document sonographic changes following unilateral nephrectomy. Normal canine renal measurements were determined sonographically prior to and following unilateral nephrectomy. These included: cortical thickness (cranial, caudal, dorsal, ventral, lateral and medial), medullary measurements (cranial and caudal) and measurements of the renal silhouette (length, height and width). The latter group of measurements was obtained to determine renal volume. Normal parameters were obtained from sixteen healthy dogs prior to nephrectomy; the unilateral nephrectomy group was comprised of eight of these animals, the remaining eight dogs were part of an allo-transplant study.¹ The mean sonographic value for the length of the kidney was 60.3 mm ± 6.4 (n = 26) while the widths and heights were 34.7 ± 3.8 (n = 27) and 27.8 mm ± 3.3 (n = 26) respectively. Renal cortical measurements were found to be smallest dorsally and ventrally on sagittal and transverse sonograms. The largest volumes were the cranial pole on sagittal scans and the lateral pole on the transverse scans. Pearson correlation coefficient for volume resulted in r values of 0.88, 0.78 and 0.72 for length, height and width (n = 25, dif = 24) respectively.     

URETERAL DUPLICATION IN A DOG

A male neutered dog of unknown age had recurrent urinary tract infection and caudal abdominal pain. Using sonography, large dilated tubular structure filled with echogenic fluid was seen extending from the left kidney to the level of the bladder neck. In an excretory urogram there was left hydronephrosis with a normal ureter. Computed tomographic evaluation of the abdomen confirmed a large tubular structure extending from the kidney with a blind ending caudally. The left kidney, ureter, and associated tubular structure were surgically removed. No connection was found between the tubular structure and the ureter. Gross evaluation and histopathologic evaluation confirmed the tubular structure to be a ureter, consistent with a diagnosis of ureteral duplication.     

ULTRASONOGRAPHIC DIAGNOSIS OF HYDRONEPHROSIS IN A COW

A ruptured right kidney was diagnosed in a cow with clinical signs of mild intermittent abdominal pain and no laboratory evidence of renal failure. Ultrasonography of the kidneys demonstrated unilateral hydronephrosis and retroperitoneal fluid. A unilateral nephrectomy using a staple instrument was performed. Ultrasonographic evaluation allowed an early and accurate diagnosis with subsequent successful surgical intervention and treatment.     

六味地黄汤对5/6肾切除大鼠NF-κB、MCP-1、Col-Ⅲ表达的影响

目的 观察六味地黄汤对5/6肾切除大鼠残肾转录因子-核因子κB（NF-κB）、单核细胞趋化因子（MCP-1）及Ⅲ型胶原（Col-Ⅲ）表达的影响,探讨六味地黄汤抑制肾间质纤维化进展的作用机制。方法 按照随机数字表法将60只SD雄性大鼠分为5组：空白组、假手术组、模型组、依那普利组、六味地黄汤组,每组12只,后三组行5/6肾切除术诱导大鼠肾衰模型,假手术组同期手术,但不损及肾脏。造模术后3 d进行灌胃干预。灌胃8周后处死各组大鼠,观察各组大鼠残肾组织形态学变化并用免疫组化法检测大鼠残肾NF-κB、MCP-1、Col-Ⅲ的表达。结果（1）观察5/6肾切除大鼠残肾组织细胞形态学,与模型组相比六味地黄汤、依那普利组可减轻大鼠肾间质损害及纤维化程度;（2）免疫组化半定量分析显示六味地黄汤组肾皮质的NF-κB、MCP-1及Col-Ⅲ表达均明显低于模型组（P<0.05）,与依那普利组差异无统计学意义（P>0.05）。结论 六味地黄汤可能通过下调NF-κB、MCP-1及Col-Ⅲ的表达,减少细胞外基质的积聚,减轻炎症反应和纤维化程度,抑制5/6肾切除大鼠肾间质纤维化进展,可作为慢性肾衰竭的辅助用药。