姜黄素对单侧输尿管梗阻大鼠肾间质纤维化的影响

目的 探讨姜黄素对单侧输尿管梗阻(UUO)大鼠模型的影响及其可能机制.方法 将30只大鼠随机分为3组,每组10只:假手术组、模型组、姜黄素组.模型组和姜黄素组行右侧输尿管接扎术,假手术组只游离不接扎.术后第14天处死各组中的大鼠,股动脉取血,检测血清肌酐、尿素氮;留取梗阻侧肾脏,Maason染色观察肾间质纤维化程度,免疫组织化学方法测定TGF-β1、CTGF的表达情况,RT-PCR技术榆测肾组织TGF-β1 mRNA、CTGF mRNA表达.结果 姜黄素降低了BUN、Scr的含量,同时姜黄素显著减少了大鼠肾间质TGF-β1、CTGF的表达,并有效改善了肾脏的病理学损伤.结论 姜黄素对单侧输尿管梗阻大鼠有较明显的保护作用,这可能与其能减少大鼠肾间质TGF-β1、CTGF的表达有关.     

Effects of renal sympathetic denervation on renal interstitial fibrosis in rats with unilateral ureteral obstruction

Objective To observe the influence of renal sympathetic denervation (RSD) on renal interstitial fibrosis and transforming growth factor beta 1(TGF-β1) and microRNA-21 (miR-21) in rats with unilateral ureteral obstruction(UUO). Methods 40 male Wistar rats were randomly divided into UUO group (A group, n=10), sham UUO group (B group, n=10), RSD+UUO group (C group, n=10) and RSD+sham UUO group (D group, n=10). Rats in A group and C group underwent unilateral ureteral ligation, while those in B group and D group underwent sham operation. Rats in C group and D group were followed by RSD. Rats were sacrificed at 21 days after the operation to evaluate the fibrosis by Masson staining. Immunohistochemical staining and Western blotting were used to detect the expressions of collagen I (COL-I), collagen Ⅲ(COL-Ⅲ) and TGF-β1 in four groups. The expression of miR-21 was detected by fluorescence in situ hybridization (FISH) and quantitative real-time PCR (RT-qPCR). Results A large amount of collagen deposition was observed in the renal interstitial area in A and C group compared to either B or D group (P＜0.05), but the change in C group was decreased significantly than that in A group (P＜0.05). Similarly, the expressions of COL-I, COL-Ⅲ, TGF-β1 and miR-21 were obviously higher in A and C group compared to either B or D group (P＜0.05), but those change in C group were decreased significantly than those in A group (P＜0.05). The above indexes were not significantly different between B group and D group (P＞0.05). Conclusion RSD may relieve the renal interstitial fibrosis in UUO rats, and down-regulate the expression of TGF-β1 and miR-21.     

Protective effect of emodin on renal interstitial fibrosis in mice of unilateral ureteral obstruction via PI3K/Akt/mTOR signaling pathway

Objective To investigate the effect and mechanism of emodin (EM) in renal interstitial fibrosis of unilateral ureteral obstruction (UUO) mice. Methods Male C57BL/6J mice were randomly divided into 4 groups, including sham operation group (n=8), UUO operation group (n=8), UUO operation+losartan (LST) group (n=8) and UUO operation+EM group (n=8). The mice in each group were ingested the suspensions by gavage for 14 days after surgery. Mice in UUO+LST and UUO+EM groups were given 10 mg?kg-1?d-1 LST and 20 mg?kg-1?d-1 EM, respectively. LST and EM were mixed with 0.5% sodium carboxymethyl cellulose. Mice in sham group and UUO group were given 0.5% sodium carboxymethyl cellulose. The mice were sacrificed at the 14th day. Interstitial fibrosis was observed by HE, Masson and PAS stain. Real-time PCR was used to detect LC3, Beclin-1 and mTOR mRNA. Protein expressions of TGF-β1, α-SMA, E-cadherin, LC3, Beclin-1, PI3K, p-Akt and mTOR were detected by Western blotting. The autophagy was observed with transmission electron microscopy in the renal tissue. Results Compared with sham mice, UUO mice at the 14th day displayed obvious renal fibrosis. Meanwhile, UUO mice had increased expressions of TGF-β1 and α-SMA (all P＜0.01), and decreased expressions of E-cadherin (P＜0.01). Their renal expressions of PI3K, p-Akt and mTOR were also raised (all P＜0.01). Compared with those in UUO group, in UUO+LST group and UUO+EM group, expressions of autophagy protein LC3 and Beclin-1 were increased (all P＜0.01), and the number of autophagic was increased. Additionally, expressions of TGF-β1 and α-SMA were reduced in UUO+LST group and UUO+EM group (all P＜0.01), while the expression of E-cadherin was increased by emodin treatment (P＜0.05). And expressions of PI3K, p-Akt and mTOR were decreased in UUO+LST group and UUO+EM group (all P＜0.05), meanwhile renal tissue fibrosis significantly reduced. Conclusions Emodin can promote autophagy, ameliorate renal interstitial fibrosis and protect renal function through PI3K/Akt/mTOR signaling pathway.     

雌激素对单侧输尿管梗阻大鼠肾间质纤维化的保护作用

目的探讨雌激素对单侧输尿管梗阻（UUO）大鼠肾间质纤维化的作用。方法雌性SD大鼠30只，随机分为4组：Ⅰ组对照组；Ⅱ组生理雌激素组；Ⅲ组低雌激素组；Ⅳ组高雌激素组。UUO术后21d处死各组大鼠，光镜观察梗阻肾组织病理变化，并分别用免疫组化和RT-PCR方法检测各组肾组织α-平滑肌肌动蛋白（α-SMA）和金属蛋白酶1组织抑制剂（TIMP-1）的表达。结果低雌激素组间质纤维化病变最明显，高雌激素组病变显著减轻（P〈0．01）。与生理雌激素组相比，低雌激素组α—SMA和TIMP-1蛋白和基因的表达增加（P〈0．05）；高雌激素组上述物质表达则减少（P〈0．05）。结论雌激素可能通过抑制α-SMA和TIMP-1的表达进而减少细胞外基质的沉积而发挥肾保护作用。     

JAK-STAT通路在小鼠单侧输尿管梗阻模型肾间质纤维化中的作用

目的 探讨Janus蛋白酪氨酸激酶-信号转导子和转录激活子(JAK-STAT)通路在小鼠单侧输尿管梗阻(UUO)模型.肾间质纤维化过程中的作用.方法 选用30只雄性Balb/c小鼠建立小鼠UUO模型(n=24)和假手术小鼠(n=6),术后第1、4、7和14天检测JAK-STAT磷酸化情况.另把18只雄性Balb/c小鼠随机分为假手术组、UUO模型组和治疗组,每组各6只.治疗组在建模前2 h开始给予选择性JAK2抑制剂AG490治疗,每天1次;模型组仅注射溶媒.术后第14天处死动物.组织学评估肾小管损伤和.肾间质纤维化程度;免疫组化检测肾脏巨噬细胞浸润和α-SMA表达;RT-PCR检测Ⅲ型胶原和单核细胞趋化蛋白(MCP)1 mRNA表达;Western印迹检测JAK2和STATl磷酸化.结果 JAK2-STAT1在UUO模型中被激活,其磷酸化水平与病情、肾小管组织学损害以及.肾间质纤维化相一致.AG490能显著抑制JAK2和STAT1的磷酸化(P＜0.01).AG490治疗显著减轻肾小管损害[(21.7±1.7)%比(49.4±1.0)%]和肾间质纤维化(1.0±0.1比2.3±0.2)、α-SMA表达(0.9±0.1比2.1±0.2)和巨噬细胞积聚[(13.3±1.6)细胞/HPF比(34.4±1.0)细胞/HPF](均P＜0.01).AG490治疗显著抑制Ⅲ型胶原和MCP-1 mRNA表达.结论 JAK-STAT信号通路在肾小管间质炎性反应和纤维化中发挥重要作用.     

内质网应激相关凋亡途径参与单侧输尿管梗阻大鼠肾间质纤维化

目的 探讨内质网应激（ERS）相关凋亡途径在单侧输尿管梗阻（UUO）大鼠肾间质纤维化发生、发展中的作用。 方法 健康雄性Wistar大鼠25只,按随机数字表法分为UUO模型组（n=18）和假手术组（n=7）,UUO模型组行左侧输尿管结扎术,假手术组仅分离输尿管不结扎,分别于术后3 d、7 d、14 d处死各组大鼠,行HE和Masson染色,观察肾脏病理变化;比色法测定肾组织羟脯氨酸（HYP）含量;免疫组化法检测α平滑肌肌动蛋白（α-SMA）;原位末端标记法（TUNEL）与DNA电泳观察肾小管间质细胞凋亡情况;RT-PCR法检测梗阻侧肾组织ERS相关分子葡萄糖调节蛋白78（GRP78）mRNA表达变化;Western印迹法分析凋亡相关蛋白半胱氨酸天门冬氨酸蛋白酶3（caspase-3）和GRP78的蛋白表达变化。 结果 与假手术组比较,UUO模型组肾脏病理改变加重,肾间质纤维化程度随梗阻时间延长逐渐加重,肾组织HYP含量显著升高（P < 0.05）,肾组织α-SMA也在肾小管间质细胞广泛表达,TUNEL染色及DNA琼脂糖凝胶电泳提示大量的肾小管间质细胞凋亡。UUO模型组GRP78 mRNA表达于术后3 d即发生显著上调,而蛋白表达在术后7 d开始出现显著变化,与假手术组差异均有统计学意义（均P < 0.01）;在此后观察期间内GRP78 mRNA和蛋白均持续高水平表达。模型组大鼠肾组织caspase-3的蛋白表达在UUO术后3 d即有显著上调（P < 0.05）,且随着梗阻时间延长进行性升高,于术后7 d、14 d增多更为显著,与假手术组差异均有统计学意义（P < 0.05）。相关分析显示GRP78蛋白表达与肾组织HYP含量和caspase-3蛋白表达均呈正相关（r = 0.657,P < 0.01;r = 0.714,P < 0.01）。 结论 UUO早期即可诱导ERS标志蛋白表达变化,触发ERS。长期ERS可诱导肾小管间质细胞凋亡;caspase-3介导的ERS相关凋亡途径可能参与了肾间质纤维化过程。     

Effect of renal fibrosis after macrophage depletion in C3-deficient unilateral ureteral obstruction mice

Objective To investigate the effect and mechanism of renal fibrosis after macrophage depletion in C3-deficient unilateral ureteral obstruction mice. Methods Renal interstitial fibrosis model was established by unilateral ureteral obstruction (UUO) in male C3-deficient mice and age-matched C57BL/6 WT mice (8-12 weeks of age). Mice were randomly divided into 4 groups, including sham operation in wild type group（WT/sham）(n=18), UUO operation in wild type group（WT/UUO）(n=18), sham operation in C3-deficient group（C3KO/sham）(n=18), and UUO operation in C3-deficient group（C3KO/UUO）(n=18). The expression of complement C3 was detected by immunohistochemical staining and renal interstitial macrophages were assessed by immunofluorescence staining. Tubulointerstitial fibrosis was observed by both HE staining and Masson staining after 14 days of UUO. Collagen accumulation and score of tubulointerstitial injury were obtained. Wild type and C3-deficient UUO mice were treated by liposome clodronate in early or late stage respectively and then interstitially infiltrated macrophages and renal fibrosis were analysed. Mice were sacrificed randomly at 3,7,14 days after UUO and obstructed kidneys were collected. Macrophage phenotype was detected by double-labeling immunofluorescence with F4/80 and iNOS for the M1, F4/80 and CD206 for the M2 macrophage subpopulation. iNOS, Arg-1 and CD206 were also detected by western blot. Results C3 deficient mice exhibited attenuated renal fibrosis, reduced collagen accumulation and tubulointerstitial injury score compared with WT mice (P＜0.01). Meanwhile, macrophage depletion in early or late stage of UUO reduced renal fibrosis in WT mice, but had no effect on C3-deficient UUO mice. Decreased accumulation of M1 macrophages and expression of iNOS, increased accumulation of M2 macrophages and expression of Arg-1, CD206 were found in C3 deficient mice compared with WT mice in early stage of UUO (P＜0.01). Conclusion Renal fibrosis is not reduced after depletion of macrophages in C3 deficient UUO mice due to the altered macrophage polarization.     

Effect of NADPH oxidases inhibition on the development of interstitial fibrosis in unilateral ureter obstruction rats

ObjectiveTo clarify whether the NADPH oxidases (NOXs) family contributed to the reactive oxygen species (ROS) production and subsequent interstitial fibrosis in unilateral ureter obstruction (UUO) rats. MethodsMale Wistar rats were randomly divided into sham operation group (n＝8), sham operation + apocynin treatment group (n＝8), UUO operation group (n＝8) and UUO operation+apocynin treatment group (n＝8). Either vehicle or apocynin (100 mg/kg per day) were given by gavage for 7 days after surgery. Rats were sacrificed at 7th day. ELISA was used to detect the activity of superoxide dismutase (SOD) and catalase (CAT), and the level of 8-iso-prostaglandin F2alpha (8-iso- PGF2α) in renal tissue. Western blotting was used to detect the protein expressions of NADPH oxidase subunit NOX2 and NOX4, α- smooth muscle actin(α-SMA), collagen I (COL-I) and the level of ERK1/ 2 phosphorylation (p-ERK1/2). ResultsUUO rats with vehicle displayed increased oxidative stress, as measured by renal tissue 8-iso-PGF2α, accompanied with increased renal expression of NADPH oxidases (NOX2, 1.5-fold and NOX4, 1.7-fold, respectively), compared with sham-operated rats (P＜ 0.05). Furthermore, vehicle treated UUO rats showed increased renal COL - I and α - SMA levels, compared with sham-operated rats (P＜0.05). ERK1/2 was also activated as detected by p-ERK1/2 expression in UUO rats with vehicle (P＜0.05). Apocynin treatment significantly decreased renal tissue 8-iso-PGF2α level and expressions of NOX2 (-28.7%) and NOX4 (-31.0%) in UUO rats, respectively, compared with vehicle treated rats (P＜0.05). And significant decrease of COL-I (-26.4%) and α-SMA expression (-80.0%) were also observed (P＜0.05). The activation of ERK1/2 in UUO rats was greatly inhibited by apocynin treatment (P＜0.01). Despite the pronounced dysregulation of pro – oxidative NOXs family, no compensatory increase of antioxidative enzyme activities occurred. ConclusionThe NOXs family contributes largely to the production of ROS and subsequent interstitial fibrosis after ureter ligation, and inhibition of the NOXs family may be a choice for preventing interstitial fibrosis.     

Wnt-7a蛋白抑制单侧输尿管梗阻小鼠肾脏上皮细胞-间充质细胞转分化

目的 观察Wnt-7a蛋白对单侧输尿管梗阻(UUO)模型小鼠肾脏纤维化的拮抗作用.方法 18只雄性C57BL/6小鼠按随机数字法分为假手术组、UUO模型组和Wnt-7a蛋白治疗组,每组6只.于术后7d处死小鼠,留取梗阻侧肾组织,行Masson染色观察肾间质纤维化程度;免疫组化染色检测肾组织E-钙黏蛋白、α-平滑肌肌动蛋白( α-SMA)、波形蛋白的表达;Western印迹法测定肾皮质中E-钙黏蛋白和 α-SMA的表达.结果 术后第7天,与假手术组相比,模型组体质量显著下降(P＜0.05),肾组织间质纤维化相对面积显著增多(P＜0.05),肾组织α-SMA及波形蛋白的表达显著增加(P＜0.05),E-钙黏蛋白表达显著减少(P＜0.05).与模型组相比,Wnt-7a蛋白治疗组体质量无明显改变(P＞0.05),肾组织间质纤维化相对面积显著减少(P＜0.05),α-SMA及波形蛋白的表达显著减少(P＜0.05),E-钙黏蛋白表达显著增高(P＜0.05).结论 Wnt-7a蛋白可以抑制肾脏上皮细胞,间充质细胞转分化的发生而减轻肾脏纤维化.     

intermedin对单侧输尿管梗阻大鼠肾间质纤维化的影响

目的 观察intermedin( IMD)对单侧输尿管梗阻(UUO)大鼠肾间质纤维化的影响.方法 雄性Wistar大鼠40只,按随机数字表法分为假手术组(n=10,行左输尿管分离术)、模型组(UUO,n=10)、氯沙坦组(n=10)和IMD组(n=10),后3组行左输尿管结扎术.各组大鼠分别于术后第14、21天随机选取5只,腹主动脉采血并留取梗阻侧肾组织之后处死.HE、Masson染色观察肾组织病理变化;比色法测定血尿素氮(BUN)、血肌酐(Scr)及新鲜肾组织羟脯氨酸(Pro)含量;免疫组化方法检测肾组织中转化生长因子β1 (TGF-β1)、IMD的表达水平,并进行半定量分析.结果 与假手术组相比,不同时间点UUO组血BUN、Scr、肾组织Pro含量及TGF-β1、IMD的阳性表达均显著升高(P＜0.05);与UUO组相比,氯沙坦组血BUN、Scr、肾组织Pro含量及TGF-β1、IMD表达均降低(P＜0.05),IMD组除IMD表达增加外,其余均降低(P＜0.05).结论 IMD可减轻UUO肾间质纤维化,改善肾功能,其机制可能与拮抗纤维化炎性介质TGF-β1有关.     

Triptolide attenuates renal interstitial fibrosis in rats with unilateral ureteral obstruction

Aim: Extracts of Tripterygium wilfordii Hook F. have been used to treat glomerulonephritis for more than 30 years in China. Most of the anti‐inflammatory and immunosuppressive activities of these extracts can be attributed to triptolide (Trip). The present study was to investigate the effect of Trip on renal interstitial fibrosis in a model of unilateral ureteral obstruction (UUO). Methods: UUO or sham‐operated rats were randomly assigned to receive mycophenolate mofetil (MMF), Trip or vehicle and were killed on days 7 and 14 after UUO or sham operation. Kidney specimens were fixed for immunohistochemistry for myofibroblasts (α‐smooth muscle actin, α‐SMA), macrophages (ED‐1), monocyte chemoattractant protein‐1 (MCP‐1) and osteopontin. Interstitial collagen deposition and amounts of transforming growth factor‐β1 (TGF‐β1) were determined by Sirius red staining and enzyme‐linked immunosorbent assay, respectively. The mRNA expression of TGF‐β1, connective tissue growth factor (CTGF), MCP‐1 and osteopontin were measured by real‐time polymerase chain reaction analysis. Results: The scores for the density of α‐SMA‐ and ED‐1‐positive cells, the staining of MCP‐1 and osteopontin, interstitial collagen deposition and amounts of TGF‐β1 were significantly reduced by MMF or Trip. MMF or Trip significantly reduced the mRNA expression of TGF‐β1, CTGF, MCP‐1 and osteopontin. Conclusion: Trip significantly attenuated tubulointerstitial fibrosis in a rat UUO model and the effect of Trip on renal fibrosis was similar to that of MMF. Trip may be useful as a potential candidate in the treatment of renal fibrosis.     

大剂量洛沙坦对单侧输尿管梗阻模型肾间质炎性反应和纤维化的影响

目的 观察血管紧张素Ⅱ受体拮抗剂洛沙坦对大鼠单侧输尿管梗阻（UUO）模型肾间质纤维化的影响，探讨大剂量能否优于常规剂量起到更多的肾脏保护作用及其可能机制。 方法 建立大鼠UUO模型术后3 d，大鼠被分为常规剂量治疗组、大剂量治疗组、手术组和假手术组。大剂量治疗组和常规剂量治疗组每天分别给予洛沙坦50 mg/kg、500 mg/kg灌胃，假手术组及手术组每天给予同体积生理盐水灌胃。观察大鼠模型第7、14和21 天鼠尾袖带血压（TCP）、尿蛋白量（24 h）、肾小管损害百分比、肾间质纤维化程度（INT）、肾间质巨噬细胞数和转化生长因子（TGF）β1 mRNA的表达。 结果 随着梗阻时间延长，手术组TCP逐渐增高，尿蛋白量（24 h）、INT、肾小管损害程度及肾间质巨噬细胞浸润显著增加，TGF-β1 mRNA含量明显增加，与其余各组相比差异有统计学意义（P < 0.05）。同时，相对于常规剂量组，大剂量治疗组能更有效的减少蛋白尿，减轻肾小管损害和肾间质纤维化，减少肾间质单核巨噬细胞浸润，显著抑制TGF-β1 mRNA表达（P < 0.05）。 结论 大剂量洛沙坦在UUO大鼠模型中较常规剂量起到了更多的肾脏保护作用，削弱炎性细胞与AngⅡ之间正反馈的恶性循环可能是其机制之一。     

支架蛋白JLP在单侧输尿管梗阻小鼠肾间质纤维化中的作用及其机制探讨

Objective To observe the effect of JLP deficiency on the progression of renal interstitial fibrosis in mice model of unilateral ureteral obstruction (UUO), and to investigate the role and underlying mechanism of JLP in the development of renal fibrosis in obstructive nephropathy. Methods jlp Wild type (jlp+/+) and jlp deficient (jlp-/-) mice were divided into four groups: jlp+/+- and jlp-/--sham-operated groups(jlp-/--sham group and jlp+/+-sham group), jlp+/+- and jlp-/--unilateral ureteral obstruction (UUO)-operated groups (jlp-/--UUO group and jlp+/+-UUO group). Mice were sacrificed at 7 days and 14 days after the operation respectively to evaluate the fibrosis by Masson staining.The expression of JLP in jlp+/+renal tissue was assayed by immunohistochemistry staining, immunofluorescence and Western blotting. Immunohistochemical staining was used to detect the expression of α-smooth muscle actin (α-SMA), collagen Ⅰ(COL-Ⅰ), collagen Ⅲ(COL-Ⅲ) and transforming growth factor-β1 (TGF-β1) in sham and UUO groups. Besides, the α-SMA, COL-Ⅰ, COL-Ⅲ, TGF-β1, p-Smad2 and p-Smad3 protein levels were also analyzed by Western blotting in four groups. Results The expression of JLP was mainly demonstrated in the renal tubules of mice. A large amount of collagen deposition was observed in the renal interstitial area in jlp-/--UUO group compared to jlp+/+-UUO group. Similarly, the expression of α-SMA, COL-Ⅰ, COL-Ⅲ and TGF-β1 was significantly increased in the kidney cortices in jlp-/-- UUO-operated groups. Meanwhile, Western blotting showed that the expression of α-SMA, COL-Ⅰ, COL-Ⅲ, and TGF-β1 protein was obviously higher in jlp-/--UUO group. Moreover, the expression of p-Smad2 and p-Smad3 protein was markedly higher in jlp-/--UUO group. Conclusion Scaffolding protein JLP is critical in preventing renal fibrosis through the inhibition of TGF-β1 expression and myo-fibroblast production.     

小鼠输尿管梗阻后肾脏间质病变及抗细胞间粘附分子1单克隆抗体的治疗作用

利用单侧输尿管梗阻制备小鼠肾间质病变模型，观察了肾间质中细胞间粘附分子－１（ＩＣＡＭ－１）及Ｍａｃ－１阳性细胞的变化，并利用抗小鼠ＩＣＡＭ－１单克隆抗体对此模型进行了治疗。结果发现在单侧输尿管梗阻后第一天，梗阻侧肾间质小血管附近及近端肾小管上即有ＩＣＡＭ－１及ＩＣＡＭ－１ｍＲＮＡ的表达，第３～５天达高峰，第７天时开始下降。与此同时，肾间质内Ｍａｃ－１阳性的巨噬细胞也于梗阻后第３天开始出现，第７天达高峰。经抗ＩＣＡＭ－１单克隆抗体治疗后，ＩＣＡＭ－１及Ｍａｃ－１的表达均延迟出现并明显减弱，肾间质病理改变亦有所减轻。     

Effect of macrophage polarization on interstitial fibrosis in mouse unilateral ureteral obstruction model

Objective To investigate the effect of macrophage polarization on tubulointerstitial fibrosis of mouse unilateral ureteral obstruction(UUO) model. Methods Twelve male C57BL/6J mice were employed, each of which with an age of 8 to 10 weeks. UUO model was established with these mice with the method of unilateral ureteral ligation. Mice were then sacrificed on the 7th and 14th day respectively after operation, and renal tissue specimens were obtained. The authors detected collagen deposition by Masson staining, and alpha smooth muscle actin (alpha SMA) as well as collagen type I (Coll-1) mRNA by real-time quantitative PCR. The authors also detected the degree of renal interstitial macrophages infiltration and expression changes of polarization by immunofluorescence staining. Results Compared with the mice that were observed on the 7th day after operation, the degree of renal interstitial fibrosis in mice observed on the 14th day after operation was comparatively serious, the difference shown by semi-quantitative results was statistically significant (P＜0.05). Moreover, mice observed on the 14th day after operation have more M2 macrophages, the difference between two groups of mice was statistically significant (P＜0.05). On the contrary, there was no statistically significant difference in the degree of M1 macrophages infiltration between these two groups of mice. Conclusions In the renal interstitial fibrosis model induced by UUO, the degree of macrophage infiltration increased significantly, mainly resulted from M2 macrophage infiltration, suggesting that M2 macrophages were involved in the formation of renal fibrosis.     

Effect of intermedin on microvascular injury of unilateral ureteral obstruction rats

Objective To observe the effect of intermedin(IMD) on microvascular injury of renal fibrosis in unilateral ureteral obstruction (UUO) rat model. Methods Seventy-two male Wistar rats were randomly divided into two groups: the sham - operation group (n=24) underwent the left ureteral dissection, the other 48 rats were made as unilateral ureteral obstruction models and sub - divided into model group(UUO, n=24) and IMD group (n=24). At the 7, 14, 21, 28 day after the operation, 6 randomly - selected rats from each of the three groups respectively were blooded by abdominal arotic and their obstructive kidneys were taken out. The renal histopathological changes were observed through HE and Masson staining, the contents of BUN, Scr and cystatin C (CysC) of the obstructive kidneys were determined, the expressions of transforming growth factor - β1 (TGF - β1), α-SMA, bone morphogenetic protein-7 (BMP-7), E-cadherin, thrombospondin 1 (TSP-1) and vascular endothelial growth factor (VEGF) were detected by RT - PCR and immunohistochemistry. Results Compared with the sham-operated group, the pathological changes of kidney in the model group showed that the degree of fibrosis was obvious, tubular interstitial damage aggravated, the levels of BUN, Scr, CysC in the model group increased (P＜0.05), the mRNA expression and protein content of TGF-β1, α-SMA, TSP-1 increased (P＜0.05), while the levels of BMP-7, E-cadherin and VEGF decreased (P＜0.05). Compared with the UUO group, renal tubular damage, interstitial fibrosis in the IMD group were lighter, the levels of BUN, Scr, CysC in the IMD group were lower (P＜0.05), the mRNA expression and protein content of TGF-β1, α-SMA,TSP-1 were down-regulated (P＜0.05), while the levels of BMP-7, E-cadherin and VEGF were up-regulated (P＜0.05). Conclusion IMD can ameliorate the renal interstitial fibrosis, and the mechanism may be related to the fact that VEGF mediated by IMD can reduce vascular injury.     

单侧输尿管梗阻大鼠肾间质纤维化的发生机制

目的 探讨单侧输尿管梗阻后肾间质纤维化的形成机制.方法 采用单侧输尿管梗阻(uniliteral ureteral obstruction UUO)致肾间质纤维化大鼠模型,将54只大鼠随机分假手术组、梗阻组.术后7天、14天、21天观察肾组织病理改变,并应用免疫组织化学方法检测肾间质核转录因子-κB(Nuclear factor-κB NF-κB)、纤溶酶原激活物抑制剂-1(plasminogen activator inhibitor-1 PAI-1)的表达.结果 随着梗阻时间延长,梗阻组肾间质PAI-1和NF-κB的表达增加,假手术组无明显变化,两组相比有显著差异(p＜0.01).结论 单侧输尿管梗阻肾间质NF-κB、PAI-1的过度表达,是形成间质纤维化的重要原因之一.     

与大鼠肾间质纤维化相关microRNA的初步研究

目的 观察microRNA(miRNA)在单侧输尿管梗阻(UUO)大鼠模型中的表达,筛选与肾间质纤维化(RIF)相关的miRNA.方法 将48只SD大鼠分为UUO组和假手术组(Sham组),时间点为术后3d、7d、14 d.Masson染色观察肾脏病理.茎环实时定量PCR检测48条miRNA在肾组织的表达.结果 术后第3天,UUO组可见灶状肾间质水肿和单个核细胞浸润 ;术后第7天,灶状间质纤维化 ;术后第14天,显著间质纤维化.Sham组肾脏未见明显病变.相对于Sham组,UUO组在术后3d、7d和14 d分别有25条、24条和21条miRNA的表达差异有统计学意义(均P＜ 0.05).其中有5条miRNA(miR-132、miR-192、miR-194、miR-29c和miR-203)在术后3、7和14 d一致上调或下调,且与肾小管间质病变程度相关(均P＜0.05).结论 在大鼠UUO模型的RIF过程中20多条miRNA存在差异表达,提示miRNA可能参与RIF的发生发展过程.miR-132、miR-192、miR-194、miR-29c和miR-203与大鼠RIF关系密切,值得进一步研究.     

小鼠单侧输尿管梗阻诱导肾脏纤维化模型中Cadherin6的表达变化

目的研究钙黏蛋白6(cadherin6,CDH6)在肾脏纤维化中表达的变化。方法建立单侧输尿管梗阻(UUO)小鼠模型,术后21天收获肾脏组织,HE染色观察肾间质纤维化程度,RT-q PCR检测肾组织FSP-1、TGF-β1、CDH6 m RNA表达量,免疫组化染色检测肾组织FSP-1、CDH6蛋白表达量;TGF-β1刺激人肾小管上皮细胞HK-2,RT-q PCR及Western Blot分别检测FN1、PAI-1、CDH6 m RNA表达量及CDH6蛋白表达量。结果 HE染色显示UUO组小鼠肾脏出现肾小管萎缩、大量基质沉积,与Sham组相比,UUO组小鼠肾脏组织FSP-1、TGF-β1、CDH6 m RNA表达量及FSP-1、CDH6蛋白表达量均上调;细胞实验结果显示,与空白对照组相比,TGF-β1诱导的HK-2细胞形态向成纤维细胞转变,FN1、PAI-1m RNA表达水平上调,CDH6 m RNA及蛋白表达水平上调。结论 TGF-β1诱导的肾小管上皮细胞CDH6及纤维化相关因子表达上调,伴随上皮细胞向间质表型转化,可能参与单侧输尿管梗阻诱导的小鼠肾脏纤维化的发展。