Activation of renal tubular epithelial cells by monocytes/macrophages and the relative mechanisms

AIM: To observe the direct effects of peripheral blood monocytes/macrophages (MO/MAC) on renal tubular epithelial cells (RTEC),and further probe into the possible mechanisms. METHODS: Conditioned medium(M-CM) of human peripheral blood MO/MAC was collected and added to HK-2,a human renal proximal tubular cell line.After incubation with M-CM for 24 hours,HK-2 cells were detected for DNA synthesis by [³H]-TdR incorporation,osteopontin (OPN) and α-smooth muscle actin (α-SMA) expression by Western blot,and fibronectin(FN) secretion by ELISA.Furthermore,anti-TGFβ₁ neutralizing antibody and interlukin-10(IL-10) were used separately to antagonize the effects of M-CM on HK-2 cells. RESULTS: ①DNA synthesis,α-SMA expression and FN secretion were all increased in HK-2 cells when incubated with M-CM.②When adding with anti-TGFβ₁ neutralizing antibody (5 mg/L) in the M-CM,the degree of upregulation of α-SMA and FN in HK-2 cells was much lower than that stimulated by M-CM alone.③M-CM added with IL-10 (20 μg/L) had a weaker ability to induce the increasing in α-SMA expression and FN excretion in HK-2 cells, compared with M-CM itself alone.M-CM from MO/MAC preincubated with IL-10 caused a lower upregulation of α-SMA expression in HK-2 cells than M-CM from non-preincubated MO/MAC. CONCLUSION: MO/MAC can directly induce proliferation,transdifferentiation and extracellular matrix secretion in RTEC.TGFβ₁ and proinflammatory cytokines secreted by MO/MAC might be involved in the aboveeffects.     

Effects of intrauterine growth restriction during late pregnancy on the ovine fetal renal function and antioxidant capacity

This study investigated the effects of intrauterine growth restriction during late pregnancy on the ovine fetal renal function and renal antioxidant capacity. Eighteen ewes pregnant were randomly divided into control group (CG, ad libitum, 0.67 MJ ME·BW^−0.75·day⁻¹, n = 6), restricted group 1 (RG1, 0.18 MJ ME·BW^−0.75·day⁻¹, n = 6), and restricted group 2 (RG2, 0.33 MJ ME·BW^−0.75·day⁻¹, n = 6). At 140 days, the fetal blood, allantoic fluid and kidney tissue were collected to determinate fetal renal function and renal antioxidant capacity. The results showed that the fetal weight, kidney weight, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), aquaporin-2 (AQP-2) and aquaporin-3 (AQP-3), and total antioxidant capacity (T-AOC) in RG1 group were decreased compared with the CG (P < 0.05), but the contents of β2-Microglobulin (β 2-MG), cystatin C (Cys-C), filtered sodium excretion fraction (FENa), malondialdehyde (MDA), and hydroxyl radical (OH) in RG1 group were increased (P < 0.05). The impaired ovine fetal renal growth, antioxidant imbalance and dysfunction of glomerulus ultrafiltration, and the renal tubules reabsorption were induced by maternal malnutrition during late pregnancy.     

鸡贮精腺上皮细胞的分离及原代培养

试验旨在探讨稳定可靠的贮精腺上皮细胞分离及原代培养方法,为研究鸡贮精机理提供细胞模型。以鸡输卵管的子宫阴道交接部组织样为材料,采用酶消化法和组织块培养法分离培养母鸡贮精腺上皮细胞,观察母鸡贮精腺上皮细胞的培养情况,比较不同细胞培养方法获得贮精腺上皮细胞的生长情况。结果表明,用胶原酶或胰酶单独消化母鸡子宫阴道交接部组织,经100目过滤后获得的贮精腺上皮细胞24 h后可贴壁,但48～72 h后细胞死亡;用胶原酶Ⅺ(0.01 g/mL)与胰酶(0.25%)先后消化母鸡子宫阴道交接部组织后再经100目过滤获得的贮精腺上皮细胞贴壁性良好,24～48 h细胞出现明显增殖,72 h后细胞增殖速度减慢,开始死亡;用组织块培养法7 d可获得鸡贮精腺上皮原代细胞,该细胞可传2～3代;用组织块培养法获得的细胞进行免疫组化试验,发现细胞表达贮精腺差异表达基因编码的NXPH1蛋白,该蛋白在培养细胞内的表达符合其分泌蛋白特性,表明组织块培养法所获细胞可用于后续研究。综上,用组织块培养法获得的鸡贮精腺上皮细胞可为研究母鸡贮精腺机制提供细胞模型。     

大鸨肾脏组织学观察

大鸨肾表面被较深的裂分成前中后三群肾叶。每个肾叶又被其表面的浅沟分成敷个肾小叶。后者由外周的皮质和基部的髓质构成,但小叶内皮质与髓质分界不明显。肾小叶彼此借小叶间静脉分开。小叶周集合管、小叶内动脉、肾小球以中央静脉为轴呈圆周排列。在小叶内,肾小体与小叶间静脉之间主要分布近曲小管,中央静脉与肾小体之间主要分布远曲小管;髓质主要分布有髓质集合管和髓袢。     

Dynamical observation of the expression of TGF-β1 and MAPK1/3 in the renal tubules of rats with diabetes

AIM: To observe the expression of transforming growth factor β₁ (TGF-β₁), MAPK_1/3 and fibronectin (FN) in the development of renal tubulointerstitial disease. METHODS: Wistar male rats were randomly divided into normal control group, diabetic group of 1week, 2 weeks, 4 weeks and 8 weeks. Diabetic model was induced by peritoneal injection of streptozotocin. Immunohistochemistry was employed to detect the expression of TGF-β₁, MAPK_1/3 and FN in the kidney. TGF-β₁ protein in the renal cortex was checked by Western blot. BG, Scr and UP were analysed by biochemical methods, and the morphological changes in renal tubulointerstitium were also examined under microscopy on sections stained with HE and PAS. RESULTS: The expression of MAPK_1/3 and FN was observed, but not the expression of TGF-β₁ in normal renal tissue. Positive staining of TGF-β₁ was observed in the renal tubulo-interstitium in 1-week diabetic group and thereafter it increased in the course of diabetes. A continuous increase in the expression of MAPK_1/3 and FN was also observed in two - week diabetic rats. Chronologically the expression of TGF-β₁,MAPK_1/3 and FN and the ratio of KW/BW were positively correlative with each other in diabetic animals except one -week diabetic rats. There was also a positive correlation between MAPK_1/3 and FN in l -week diabetic rats. CONCLUSION: Our data suggest that TGF-β₁ appears in the renal tubulointerstitium in early period of diabetes and then its signal is mediated by MAPK_1/3 cascades to accelerate production of FN ,and in turn leads to renal hypertrophy and tubulointerstitial fibrosis.     

Reducing endogenous estrogen during development alters hormone production by porcine Leydig cells and seminiferous tubules

High levels of estrogen produced by boar testes and the presence of estrogen receptors in both interstitial and tubular compartments are consistent with a direct role for estrogen in regulation of testicular cell function. This study investigated the importance of estrogen on hormone production by Leydig cells and seminiferous tubules in the developing boar. Thirty-six 1-week-old littermate pairs of boars were treated weekly with vehicle or 0.1 mg/kg BW Letrozole, an aromatase inhibitor, until castration at 2, 3, 4, 5, 6, 7, or 8 months. Tissue was collected and Leydig cells and seminiferous tubules were isolated. In a separate study, five untreated boars (ages 1.5-4 months) were castrated and Letrozole was added in vitro to Leydig cell and seminiferous tubule cultures. Leydig cells were cultured for 24h with and without porcine LH. Media were assayed for estradiol (E(2)) and testosterone (T) concentrations by RIA. Seminiferous tubules were cultured for 4h with and without porcine FSH; media were assayed for E(2) and immunoreactive inhibin (INH). In vivo aromatase inhibition decreased basal E(2) and increased basal T production by cultured Leydig cells. Basal seminiferous tubule production of E(2) but not INH was reduced. Decreasing estrogen synthesis in vivo did not alter LH-induced Leydig cell E(2) production or FSH-induced seminiferous tubule INH production. INH production decreased with advancing age regardless of treatment. In conclusion, in vivo aromatase inhibition altered baseline steroid production by cultured Leydig cells and seminiferous tubules but had little effect on response to gonadotropins.     

大鸨肾脏组织学观察

大鸨肾表面被较深的裂分成前中后三群肾叶。每个肾叶又被其表面的浅沟分成敷个肾小叶。后者由外周的皮质和基部的髓质构成,但小叶内皮质与髓质分界不明显。肾小叶彼此借小叶间静脉分开。小叶周集合管、小叶内动脉、肾小球以中央静脉为轴呈圆周排列。在小叶内,肾小体与小叶间静脉之间主要分布近曲小管,中央静脉与肾小体之间主要分布远曲小管;髓质主要分布有髓质集合管和髓袢。     

Spontaneous Accumulation of Globotriaosylceramide (Gb3) in Proximal Renal Tubules in an ICR Mouse

This report describes spontaneous cytoplasmic vacuolation in the proximal renal tubules of a 7-week-old male ICR [Crlj:CD1(ICR)] mouse. The contents of vacuoles were positively stained with periodic acid-Schiff (PAS) and Sudan black, and the membranes were positive on immunohistochemical staining for lysosomal-associated membrane protein-2 (LAMP-2), a marker of lysosomal membrane. Electron microscopy revealed electron-dense lamellar bodies in the proximal tubular epithelial cells. These histopathological features are similar to those in α-galactosidase A-deficient mice, in which globotriaosylceramide (Gb3), a glycosphingolipid, accumulates in lysosomes. When we performed immunohistochemical staining for Gb3, the contents of vacuoles were positively stained. From these results, spontaneous cytoplasmic vacuolation in the proximal renal tubules in the mouse was identified as lysosomal accumulation of Gb3.