Peroxisome Proliferator-activated Receptor-γ Agonist Pioglitazone Fails to Attenuate Renal Fibrosis Caused by Unilateral Ureteral Obstruction in Mice

Renal tubulointerstitial fibrosis is the common ending of progressive renal disease. It is worth developing new ways to stop the progress of renal fibrosis. Peroxisome proliferator-activated receptor-γ（PPARγ） agonists have been studied to treat diabetic nephropathy, cisplatin-induced acute renal injury, ischemia reperfusion injury and adriamycin nephropathy. In this study, unilateral ureteral obstruction（UUO） was used to establish a different renal fibrosis model. PPARγ agonist pioglitazone was administrated by oral gavage and saline was used as control. At 7th and 14 th day after the operation, mice were sacrificed for fibrosis test and T lymphocytes subsets test. Unexpectedly, through MASSON staining, immunohistochemistry for α-SMA, and Western blotting for α-SMA and PDGFR-β, we found that pioglitazone failed to attenuate renal fibrosis in UUO mice. However, flow cytometry showed that pioglitazone down-regulated Th1 cells, and up-regulated Th2 cells, Th17 cells and Treg cells. But the Th17/Treg ratio had no significant change by pioglitazone. Real-time PCR results showed that TGF-β and MCP-1 had no significant changes, at the same time, CD4＋ T cells associated cytokines were partially regulated by pioglitazone pretreatment. Taken together, pioglitazone failed to suppress renal fibrosis progression caused by UUO.

Expression of BMP-7 and its receptors in renal tubolo-interstitial fibrosis induced with unilateral ureteral obstruction in rats

Objective:To study the changes of the expression of bone morphogenetic protein-7(BMP-7) and its receptors(BMPR-Ⅱ,ALK2,ALK3 and ALK6) in the renal tubulointerstitial fibrosis induced with unilateral ureteral obstruction in rats. Methods: Sixty Wistar male rats were divided randomly into the normal control,sham-operation and unilateral ureteral obstruction(UUO) groups and the rats were killed on the 1st,3rd,7th and 14th postoperative days respectively.The mRNA level of BMP-7,BMPR-Ⅱ,ALK2,ALK3 and ALK6 was determined with RT-PCR.The site and level of protein expression of BMP-7 were observed with immunohistochemical staining. Results: The mRNA level of BMP-7,BMPR-Ⅱ,ALK2 and ALK3 was significantly decreased in the rats of UUO group than in those of the sham-operation group but the mRNA level of ALK6 showed no obvious changes in all the rats.Immunohistochemical staining revealed that the protein of BMP-7 was mainly expressed in the renal tubules and interstitial tissue of the kidneys in normal rats but it was decreased gradually along with the unilateral ureteral obstruction. Conclusion: It is found that the loss of BMP-7 and its receptors including BMPR-Ⅱ,ALK2 and ALK3 occurs in the early phase of renal tubulointerstitial fibrosis before the appearance of other pathological changes in the kidney and may play an important role in the occurrence and progress of renal tubulointerstitial fibrosis.

Role of BMP-7 and TGF-β_1 expression in renal tubulo-interstitial lesion of Wistar rats induced by unilateral ureteral obstruction

Objective: To explore the role of bone morphorgenetic protein-7 （BMP-7） in the renal tubulo-interstitial lesions induced by unilateral ureteral obstruction （UUO）. Methods: Sixty Wistar rats were equally and randomly divided into normal control, sham operation and UUO groups, and respectively sacrificed on the 1st, 3rd, 7th, 14th day after the time of UUO operation. The mRNA levels of BMP-7 and TGF-β1 in the renal tissues were examined by RT-PCR. The expression sites and levels of BMP-7 and TGF-β1 proteins were detected by immunohistochemistry staining. Results:Compared to control groups, the level of BMP-7 mRNA was significantly decreased, but that of TGF-β1 mRNA was significantly increased in UUO rats. Immunohistochemistry staining indicated that BMP-7 mainly expressed in the renal tubules and interstitum, rarely in the glomeruli. In UUO rats, the expression of BMP-7 protein was decreased, but that of TGF-β1 was increased in an obstruction dependent manner. Conclusion:The down- regulation of BMP-7 is observed in the early phase of fibrotic process of the renal interstitium, suggesting it may be involved in the formation and development of the tubulo-interstitial lesions.

The effect of unilateral ureteral obstruction on the adrenal function in a rat model

【Objective】To explore the effect of unilateral ureteral obstruction on the adrenal function in a rat model.【Methods】36 SD rats were randomly divided into two groups:unilateral ureteral obstruction(UUO)model was employed,and sham-operated rats were used as control.28 days after surgery,all the rats were submitted to maximum exercise tolerance test(METT),and the maximum aerobic velocity was observed.At the day 0,day 28,and after METT,the serum levels of adrenaline,noradrenaline and corticosterone in rats were detected by ELISA.【Results】1 The maximum aerobic velocity in UUO model rats was lower than that in control rats(P<0.05).2 At the beginning(day 0),there no significant difference were found in the levels of adrenaline,noradrenaline and corticosterone between two groups.3 28 days after surgery,the levels of adrenaline,noradrenaline and corticosterone in UUO model rats were slightly higher than those in control rats,but the differences were not significant(P>0.05).4After METT,the levels of adrenaline and corticosterone in UUO model rats were lower than those in control rats significantly(P<0.05).The level of noradrenaline in UUO model rats was higher than that in control rats,but there is no significant difference between two groups(P>0.05).【Conclusions】Unilateral ureteral obstruction resulted in the decrease of exercise tolerance in rats,which may contribute to the inhibition of the adrenal secretion.

Huoxue Jiedu Huayu recipe(活血解毒化瘀方)inhibits macrophage-secreted vascular endothelial growth factor-a on angiogenesis and alleviates renal fibrosis in the contralateral kidneys of unilateral ureteral obstruction rats

OBJECTIVE:To elucidate the mechanism by which Huoxue Jiedu Huayu recipe(活血解毒化瘀方,HJHR)regulates angiogenesis in the contralateral kidney of unilateral ureteral obstruction(UUO)rats and the mechanism by which it reduces of renal fibrosis.METHODS:Male Wistar rats were randomly divided into 4 groups:the sham group,UUO group(180 d of left ureter ligation),UUO plus eplerenone(EPL)group,and UUO plus HJHR group.After 180 d of oral drug administration,blood and contralateral kidneys were collected for analysis.Angiogenesis-and fibrosis-related indexes were detected.RESULTS:HJHR and EPL improved structural damage and renal interstitial fibrosis in the contralateral kidney and reduced the protein expression levels ofα-smooth muscle actin(α-SMA),vimentin and collagen I.Moreover,these treatments could reduce the expression of vascular endothelial growth factor-A(VEGFA)by inhibiting the infiltration of macrophages.Furthermore,HJHR and EPL significantly reduced the expression of CD34 and CD105 by downregulating VEGFA production,which inhibited angiogenesis.Finally,the coexpressions of CD34,CD105 andα-SMA were decreased in the HJHR and EPL groups,indicating that endothelial-to-mesenchymal transition was inhibited.CONCLUSIONS:These findings confirm that HJHR alleviates contralateral renal fibrosis by inhibiting VEGFAinduced angiogenesis,encourage the use of HJHR against renal interstitial fibrosis and provide a theoretical basis for the clinical management of patients with CKD.

Trabecular bone deterioration at the greater trochanter ol mice with unilateral obstructive nephropathy

Our previous study showed the early molecular responses of bone in response to obstructive nephropathy in a unilateral ureteral obstruction （UUO） mouse model. Here, we addressed the changes in trabecular bone properties at greater trochanter, the proximal and the distal metaphysis of femur in UUO mice. The male mice were subjected to UUO （n= 10） or sham operation （n= 10）. All mice were killed on day 7 after the surgical operation. The micro-computed tomography （micro-CT） analysis for different femoral trabecular bone sites demonstrated pathological alterations of trabecular bone mass and micro-networks at greater trochanter as shown by decreases in bone mineral density/bone volume （P〈O.05） and trabecular number （P〈O.05） and increases in trabecular separation （P〈O.01） and bone surface/bone volume （P〈O.05） in UUO mice. The present study demonstrates that UUO-induced unilateral obstructivenephropathy has markedly detrimental effects on the trabecular trochanter of the femur.

Ginsenoside Rb1,a Panoxadiol Saponin against Oxidative Damage and Renal Interstitial Fibrosis in Rats with Unilateral Ureteral Obstruction

Objective:To investigate the possible protective effect and mechanism of ginsenoside Rb1 against oxidative damage and renal interstitial fibrosis on rats with unilateral ureteral obstruction(UUO). Methods:In total,80 male rats were randomly divided into 4 groups,20 in each group:the sham operated group (SOR),UUO group,UUO with ginsenoside Rb1 treatment group(treated with intraperitoneal injection of 50 mg/ kg daily) and UUO with Losartan treatment group(as the positive control,treated with 20 mg/kg by gastr...

Arterially transplanted mesenchymal stem cells in a mouse reversible unilateral ureteral obstruction model： in vivo bioluminescence imaging and effects on renal fibrosis

Background Chronic kidney disease （CDK） is a worldwide health problem, but there is currently no effective treatment that can completely cure this disease. Recently, studies with mesenchymal stem cells （MSCs） on treating various renal diseases have shown breakthroughs. This study is to observe the homing features of MSCs transplanted via kidney artery and effects on renal fibrosis in a reversible unilateral ureteral obstruction （R-UUO） model. Methods Thirty-six Balb/c mice were divided into UUO group, UUO-MSC group, and sham group randomly, with 12 mice in each group. The MSCs had been infected by a lentiviral vector to express stably the luciferase reporter gene and green fluorescence protein genes （Luc-GFP-MSC）. Homing of MSCs was tracked using in vivo imaging system （IVIS） 1, 3, 14, and 28 days after transplantation. Imaging results were verified by detecting GFP expression in frozen section under a fluorescence microscope. E-cadherin, α-SMA, TGF-β1, and TNF-α mRNA expression in all groups at 1 and 4 weeks after transplantation were analyzed by quantitative PCR. Results Transplanted Luc-GFP-MSCs showed increased Luciferase expression 3 days after transplantation. The expression decreased from 7 days, weakened thereafter and could not be detected 14 days after transplantation. Quantitative PCR results showed that all gene expressions in UUO group and UUO-MSC group at 1 week had no statistical difference, while at 4 weeks, except TGF-β expression （P〉0.05）, the expression of E-cadherin, α-SMA, and TNF-α in the above two groups have statistical difference （P〈0.01）. Conclusion IVIS enables fast, noninvasive, and intuitive tracking of MSC homing in vivo. MSCs can be taken home to kidney tissues of the diseased side in R-UUO model, and renal interstitial fibrosis can be improved as well.

六味地黄汤对UUO大鼠肾组织β-catenin及EMT的影响

目的探讨六味地黄汤(Liuwei Dihuang decoction,LWDHD)对单侧输尿管梗阻(unilateral ureteric obstrutction,UUO)大鼠肾组织中上皮钙黏蛋白E(E-cadherin)、β-连环蛋白(β-catenin)、α-平滑肌肌动蛋白(α-SMA)的表达、肾脏组织病理变化及上皮-间质转化(epithelial-mesenchymal transformation,EMT)的影响。方法SPF级SD大鼠48只随机分成假手术组(Sham)、模型组(UUO)、LWDHD低、中、高组(LWDHD 3.375、6.75、13.5 g·kg^(-1))、依那普利组(Enalapril)10 mg·kg^(-1),每组8只。采用HE、Masson染色法观察6组大鼠患侧肾组织的病理变化和胶原纤维沉积情况;免疫组化及Western blot检测各组大鼠肾组织中上E-cadherin、β-catenin、α-SMA蛋白的表达情况。结果与Sham组比较,UUO组大鼠患侧肾组织中肾小球硬化,肾小管扩张,间质纤维化明显。β-catenin、α-SMA蛋白表达增加,E-cadherin蛋白表达降低(P<0.05);与UUO组相比,LWDHD组和Enalapril组中大鼠肾组织病理改变和胶原沉积明显改善,β-catenin、α-SMA蛋白表达降低,E-cadherin蛋白表达增加(P<0.05)。结论LWDHD可能通过下调β-catenin的表达,进而抑制EMT,发挥缓解肾纤维化的作用。

调控Nrf2/GPX4信号通路抑制铁死亡并缓解UUO小鼠肾纤维化

目的:评估单侧输尿管梗阻(UUO)小鼠模型中调控核因子E2相关因子2(Nrf2)/谷胱甘肽过氧化物酶4(GPX4)信号通路抑制铁死亡及肾脏纤维化的作用,检测慢性肾脏病(CKD)患者肾脏组织相关铁死亡及纤维化指标的表达情况。方法:收集9例CKD患者及9例非CKD患者肾穿标本石蜡切片,免疫组化法检测GPX4、Nrf2和α-平滑肌肌动蛋白(α-SMA)的表达水平。动物实验中,25只6~8周龄雄性C57BL/6小鼠采用简单随机抽样方法分为5组:假手术组、UUO模型组、UUO+liproxstatin-1(Lip-1;铁死亡抑制剂)组、UUO+萝卜硫素(SFN;Nrf2激动剂)组、UUO+厄贝沙坦(IRB)组,每组5只。正常饲料喂养1周,后3组分别腹腔注射Lip-1(10 mg·kg^(−1)·d^(−1))、SFN(25 mg·kg^(−1)·d^(−1))和IRB(20 mg·kg^(−1)·d^(−1))。取血清标本测定血清肌酐和尿素氮;肾脏组织石蜡包埋后行HE、Masson和Sirius red染色观察肾脏病理改变;Western blot、实时荧光定量PCR和免疫组化法检测小鼠肾脏组织Nrf2、GPX4、溶质载体家族7成员11(SLC7A11)、纤连蛋白(Fn)、I型胶原(Col-I)、α-SMA和NADPH氧化酶4(NOX4)的表达。结果:免疫组化结果显示,CKD患者肾脏组织石蜡切片GPX4和Nrf2表达低于非CKD患者,而α-SMA表达高于非CKD患者。动物实验中,与UUO组相比,3组用药组肾功能有改善,肾脏纤维化程度及相关指标表达减少;Lip-1和SFN用药组肾脏组织中Nrf2、GPX4和SLC7A11表达水平高于UUO组(P<0.05),NOX4表达水平低于UUO组(P<0.05)。结论:CKD患者及UUO小鼠肾脏纤维化及铁死亡显著增加;调控Nrf2/GPX4信号通路可抑制铁死亡并有效减轻UUO小鼠肾脏纤维化。

活血解毒化瘀方通过MR/NLRP3通路抑制UUO 6个月大鼠对侧肾脏血管平滑肌细胞焦亡的机制研究

目的观察活血解毒化瘀方对单侧输尿管梗阻(Unilateral ureteral obstruction,UUO)6个月大鼠模型对侧肾脏血管平滑肌细胞(Vascular smooth muscle cell,VSMC)焦亡的干预作用,探讨活血解毒化瘀方对慢性肾脏病(Chronic kidney disease,CKD)的部分治疗机制。方法40只雄性Wistar大鼠随机分为假手术(Sham)组、单侧输尿管梗阻(UUO)组、依普利酮治疗(EPL)组和活血解毒化瘀方干预(HJHR)组(n=10)。术后EPL组给予依普利酮,HJHR组给予中药。6个月后摘取右侧肾脏。通过HE染色和Masson染色评估对侧肾脏病理改变。免疫组化检测含半胱氨酸的天冬氨酸蛋白水解酶1(Cysteinyl aspartate specific proteinase1,Caspase-1)、白细胞介素1β(Interleukin-1β,IL-1β),核因子κappa-B,p65(Nuclear Factorκappa-B,NF-κB,p65)和血清和糖皮质激素诱导的激酶1(Serum and glucocorticoid-induced kinase 1,SGK1)的表达。通过TUNEL染色观察肾脏血管平滑肌细胞的DNA损伤情况。体外实验将大鼠血管平滑肌细胞随机分为4组:空白对照(CON)组、醛固酮刺激(ALD)组、醛固酮加依普利酮治疗(EPL)组和醛固酮加活血解毒化瘀方干预(HJHR)组。ALD组给予醛固酮刺激24 h,EPL组和HJHR组在醛固酮刺激前分别给予依普利酮和10%大鼠含中药血清预处理。通过流式细胞术检测细胞凋亡率。通过免疫荧光观察细胞膜中GSDMD蛋白(Gasdermin D,GSDMD)的穿孔和核受体亚家族3C组成员2(Nuclear Receptor subfamily 3 group C member 2,NR3C2)、编码盐皮质激素受体(Mineralocorticoid receptor,MR)的核转位。通过蛋白免疫印迹法检测炎症和细胞焦亡相关信号分子的蛋白质表达。通过实时荧光定量PCR(Quantitative real-time PCR,qRT-PCR)检测细胞焦亡信号通路相关分子的信使RNA(Messenger RNA,mRNA)表达。结果与Sham组比较,UUO组肾脏血管及血管平滑肌细胞的DNA损伤加重,Caspase-1、IL-1β、SGK1、NF-κB的表达增高;与UUO组比较,EPL组和HJHR组肾脏血管及血管平滑肌细胞的DNA损伤有所减轻,Caspase-1、IL-1β、SGK1和NF-κB的表达降低。与CON组比较,ALD组出现大鼠血管平滑肌细胞焦亡及细胞DNA损伤,NR3C2、NOD样受体热蛋白结构域相关蛋白3(Nod-like receptor protein 3,NLRP3)、Caspase-1、GSDMD、IL-1β、SGK1、NF-κB的表达上调;与ALD组比较,EPL组和HJHR组细胞焦亡及细胞DNA损伤减轻,NR3C2、NLRP3、Caspase-1、GSDMD、IL-1β、SGK1、NF-κB的表达降低。结论活血解毒化瘀方通过抑制MR/NLRP3通路的激活,抑制血管平滑肌细胞焦亡,从而减轻UUO对侧肾脏血管损伤。

基于JAK/STAT3通路探讨补肾活血方改善单侧输尿管梗阻大鼠炎症和肾纤维化

目的:观察补肾活血方(BSHXF)对单侧输尿管梗阻大鼠(unilateral ureteral obstruction,UUO)肾纤维化和炎症的治疗效果,揭示其抗炎及延缓肾纤维化的机制。方法:32只8周龄雌性SD大鼠根据随机数字表分为假手术组(Sham组)、单侧输尿管梗阻组(UUO组)、补肾活血方低剂量组(BSHXF-L组)和补肾活血方高剂量组(BSHXF-H组)。造模后第2天开始,连续灌胃干预21 d,BSHXF-L组和BSHXF-H组给予相应剂量补肾活血方汤剂,Sham组和UUO组给予与BSHXF组等体积的生理盐水。干预结束后,记录24 h尿量,全自动生化分析仪检测血肌酐、尿素氮、尿蛋白、尿肌酐和肾功能指数;采用HE和Masson染色检测肾脏病理损伤和纤维化程度;通过免疫组化染色分析各组TGF-β和TNF-α;Western blotting检测各组TGF-β_(1)、α-SMA、JAK、STAT3、p-STAT3、TNF-α、Caspase-3和Bax的蛋白表达情况。结果:与UUO模型组相比,BSHXF-L组和BSHXF-H组能明显明显改善UUO大鼠的肾功能,降低大鼠肾组织中TGF-β_(1)和α-SMA蛋白表达水平,抑制JAK、STAT3和p-STAT3表达,减少炎症细胞浸润和纤维组织以及胶原的增生,显著降低大鼠的肾脏指数和TNF-α等炎症指标。结论:补肾活血方可通过抑制JAK/STAT3通路激活,改善UUO大鼠肾功能,减轻其肾脏炎症水平、肾脏指数,延缓肾纤维化进程。

NLRP3炎症小体在大鼠单侧输尿管梗阻引起肾间质纤维化中的作用及其机制

目的:探讨核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎性小体在大鼠单侧输尿管梗阻(UUO)模型肾间质纤维化中的作用,并阐明其可能的作用机制。方法:健康雄性Wistar大鼠30只随机分为假手术组(n=6)和UUO组(n=24),假手术组大鼠仅分离输尿管不结扎,UUO组分别于术后3、7和14 d处死大鼠,并按照处理时间分为UUO 3 d组(n=8)、UUO 7 d组(n=8)和UUO 14 d组(n=8)。HE染色和Masson染色观察各组大鼠肾组织病理形态表现,试剂盒检测各组大鼠肾组织中丙二醛(MDA)水平、超氧化物歧化酶(SOD)活性和羟脯氨酸(HYP)水平,免疫组织化学法检测各组大鼠肾组织中α-平滑肌肌动蛋白(α-SMA)和转化生长因子β1(TGF-β1)蛋白表达水平,Western blotting法检测各组大鼠肾组织中NLRP3蛋白表达水平。结果:HE染色,UUO组大鼠出现明显肾小管扩张,肾间质水肿和增宽,可见较多炎症细胞浸润,部分肾小管腔内可见脱落的上皮细胞。与假手术组比较,UUO 3 d组、UUO 7 d组和UUO 14 d组大鼠HE染色肾间质纤维化评分均明显升高(P<0.05);与UUO 3 d组和UUO 7 d组比较,UUO 14 d组大鼠HE染色肾间质纤维化评分明显升高(P<0.05)。Masson染色,UUO组大鼠肾间质炎症细胞浸润明显,可见明显纤维组织增生;随UUO作用时间增加,大鼠部分肾小管消失,肾间质明显增宽,胶原沉积逐渐增多,皮髓交界处胶原沉积程度更加明显。与假手术组比较,UUO 3 d组、UUO 7 d组和UUO 14 d组大鼠Masson染色肾间质纤维化评分均明显升高(P<0.05);与UUO 3 d组和UUO 7 d组比较,UUO 14 d组大鼠Masson染色肾间质纤维化评分明显升高(P<0.05)。与假手术组比较,UUO 3 d组、UUO 7 d组和UUO 14 d组大鼠梗阻侧肾组织中MDA水平均明显升高(P<0.05),SOD活性明显降低(P<0.05)。与假手术组比较,UUO 3 d组、UUO 7 d组和UUO 14 d组大鼠梗阻侧肾组织中HYP水平均明显升高(P<0.05);与UUO 3 d组比较,UUO 14 d组大鼠梗阻侧肾组织中HYP水平明显升高(P<0.05)。免疫组织化学法,与假手术组比较,UUO 3 d组、UUO 7 d组和UUO 14 d组大鼠肾组织中α-SMA蛋白表达水平明显升高(P<0.05);与UUO 3 d组和UUO 7 d组比较,UUO 14 d组大鼠肾组织中α-SMA蛋白表达水平均明显升高(P<0.05);与假手术组比较,UUO 3 d组、UUO 7 d组和UUO 14 d组大鼠肾小管上皮细胞和肾小管间质组织中TGF-β1蛋白表达水平明显升高(P<0.05);与UUO 3 d组比较,UUO 14 d组大鼠肾小管上皮细胞和肾小管间质组织中TGF-β1蛋白表达水平明显升高(P<0.05)。Western blotting法,与假手术组比较,UUO 7 d组和UUO 14 d组大鼠肾组织中NLRP3蛋白表达水平均明显升高(P<0.05)。结论:NLRP3炎症小体在UUO大鼠肾纤维化过程中发挥重要作用,其作用机制与氧化应激增加和TGF-β1蛋白表达水平升高有关。

青蒿琥酯对UUO模型大鼠肾脏的抗炎作用及其机制研究

目的观察青蒿琥酯能否有效拮抗肾脏病理性炎症过程的发生以及对肾脏疾病的治疗作用。方法将雄性Wistar大鼠60只随机分为5组,包括假手术组(SOR组)、大剂量青蒿琥酯组(HDA组)、小剂量青蒿琥酯(SDA组)、氯沙坦钾组(ARB组)、UUO模型对照组(UUO组)。从模型建立后24h开始灌胃:其中SDA、HDA组分别给予青蒿琥酯25mg·kg-1·d-1、50mg·kg-1·d-1灌胃,ARB组给予氯沙坦钾20mg·kg-1·d-1灌胃,大鼠术后3、7d收集24h尿、血、肾组织标本。行一般生化检查及病理染色,电镜观察细胞器改变情况。免疫组化法检测肾组织中磷酸化NF-Kappa B,实时荧光定量PCR检测Smad7和IκB-α的mRNA表达。结果 (1)UUO组术后3、7d血肌酐水平高于其他各组(P<0.05),各治疗组不同时间点血肌酐水平差异无统计学意义(P>0.05);(2)磷酸化NF-KappaBP56阳性细胞在成模后3d开始增加,UUO组表达显著高于对照组(P<0.05),各治疗组显著少于UUO组(P<0.05);(3)RT-PCR检测Smad7 mRNA在UUO组表达相同时间点较SOR组略有上升,各治疗组Smad7亦较SOR组有所上升,SOR组IκB-αmRNA显著高于各实验组(P<0.05),各药物治疗组IκB-α mRNA减低程度弱于UUO组(P<0.05)。结论青蒿琥酯可以减轻肾间质炎性损伤,其机制可能和上调Samd7表达及减少IκB-α mRNA降解有关。

安体舒通和缬沙坦对UUO大鼠肾脏的抗氧化保护作用

目的探讨醛固酮受体拮抗剂安体舒通和血管紧张素Ⅱ-1型受体拮抗剂(AT1Ra)缬沙坦对单侧输尿管梗阻(UUO)模型肾间质纤维化的抗氧化保护作用。方法Wistar大鼠行左侧输尿管结扎术,分为UUO模型组(n=11),安体舒通治疗组(US,n=12),缬沙坦治疗组(UV,n=11)和安体舒通+缬沙坦治疗组(USV,n=10),同时设假手术对照组(n=7)。术后第14天处死各组大鼠,进行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),丙二醛(MDA)和超氧化物歧化酶(SOD)含量;免疫组织化学方法测定α-平滑肌肌动蛋白(α-SMA)的表达。结果UUO组与其它组大鼠比较,肾组织HYP和MDA含量明显升高,SOD含量显著下降,肾脏病理改变加重,肾组织α-SMA的表达显著增加(P<0.05);各用药组与UUO组大鼠比较,肾组织HYP和MDA含量明显降低,SOD含量升高,肾间质纤维化显著减轻,肾组织α-SMA的表达显著减少(P<0.05)。结论安体舒通联合缬沙坦能减少单侧输尿管梗阻肾组织脂质过氧化物的产生,增加抗氧化酶的含量,从而显著改善UUO大鼠氧化应激所致的肾间质纤维化。

加味六味地黄汤对UUO大鼠肾组织TGF-β1及EMT的影响

目的:观察加味六味地黄汤对单侧输尿管梗阻(UUO)大鼠肾组织TGF-β1、E-钙黏素(E-Cadherin)、α-平滑肌肌动蛋白(α-SMA)的影响。方法:健康成年SD大鼠90只随机分为假手术组、模型组、中药组3组。假手术组和模型组给予生理盐水(2ml/只/d)灌胃;中药组给予加味六味地黄汤组灌胃(2 ml/d)。检测各组大鼠肾功能、肾脏病理及肾组织TGF-β1、E-Cadherin和α-SMA蛋白的表达。结果:与假手术组相同时间点比较,模型组大鼠BUN和Scr水平、小管间质半定量评分、TGF-β1、α-SMA蛋白表达均显著增高,E-Cadherin蛋白下调,差异均有统计学意义(P<0.01);与模型组大鼠相同时间点比较,加味六味地黄汤显著降低BUN和Scr水平,小管间质半定量评分、TGFβ1、α-SMA蛋白表达(P<0.05),显著上调E-Cadherin蛋白表达(P<0.05)。结论:加味六味地黄汤可能通过抑制TGF-β1,影响肾小管上皮细胞转分化,发挥抗肾间质纤维化作用。

UUO模型大鼠肾间质纤维化动态进展及α-SMA、TGF-β_1和VDR表达变化

目的观察单侧输尿管梗阻(UUO)模型大鼠肾小管间质纤维化动态进展,研究该病理过程中肾脏纤维化相关蛋白表达的变化。方法 32只清洁级SD大鼠随机分为假手术组(n=16)和UUO模型组(模型组,n=16)。两组大鼠分别于术后(模型组建模后)第2、5、9、14天分批(n=4)处死,留取手术侧(模型组梗阻侧)肾脏组织。分别采用HE和Masson染色、免疫组织化学染色及Western blotting等方法,评价肾小管间质纤维化损伤程度并检测肾脏组织α平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(FN)、维生素D受体(VDR)及转化生长因子β1(TGF-β_1)等相关蛋白的表达。结果肾脏组织形态学观察显示,随着输尿管梗阻时间的延长,肾小管间质纤维化程度逐步加重。免疫组织化学染色显示,模型组大鼠建模后各时间点肾间质α-SMA和FN阳性面积百分比均显著高于假手术组(P<0.05),且随着梗阻时间的延长逐渐升高,至建模后第14天时达到高峰。Western blotting分析表明,模型组大鼠建模后各时间点α-SMA和TGF-β_1相对表达量均显著高于假手术组(P<0.05),而VDR相对表达量显著低于假手术组(P<0.05);建模后第2天,模型组大鼠肾脏组织α-SMA相对表达量已显著增加;建模后第14天,模型组α-SMA和TGF-β_1相对表达量分别增高至假手术组的12.7倍和8.8倍,而VDR相对表达量下降至假手术组的3%。结论 UUO模型大鼠建模后第14天可出现显著的肾间质纤维化;建模早期肾间质中α-SMA表达增加提示肾间质成纤维细胞的活化;VDR表达的进行性减少提示其与肾间质纤维化有关。

大豆异黄酮对UUO大鼠TGF-β_1的抑制作用

目的 探讨大豆异黄酮对单侧输尿管梗阻 (unilateralureteralobstruction ,UUO)大鼠梗阻肾脏肾间质纤维化的防治作用及机制。方法 观察大鼠结扎侧肾脏的病理改变 ,应用HE和PAS染色观测肾间质容量 ,应用RT PCR法检测肾组织内TGF β1mRNA的表达。结果 异黄酮组梗阻肾肾间质容量明显低于对照组 ;异黄酮组TGF β1mRNA表达明显低于对照组 ,统计学分析差异显著 (P <0 0 5 ) ,异黄酮组肾间质纤维化程度轻于对照组。结论 大豆异黄酮对UUO大鼠梗阻肾肾间质纤维化有一定的抑制作用 ,其机制与大豆异黄酮可抑制TGF β1mRNA的表达有关。

CHIP调节UUO再通大鼠肾组织TGF-β1/Smads信号通路的研究

目的:利用基因芯片研究Carboxyl terminus of Hsc70-interacting protein(CHIP)对肾间质纤维化TGF-β1/Smads信号通路的抑制性调节。方法:将18只SPF级SD大鼠中6只行unilateral ureteral occlusion(UUO)模型,6只植入弹性管脂肪垫加压梗阻,7天后再通定为reverse of unilateral ureteral occlusion(RUUO)组,6只UUO在7天处死留取左肾组织,另外6只为假手术组。6只RUUO大鼠在再通7天后处死,留取左侧肾组织,运用West-ern blotting检测肾组织CHIP的表达,免疫组化检测肾组织中α-SMA、CD68的阳性表达面积,定制芯片研究TGF-β1/Smads信号通路的基因变化。结果:UUO组TGF-β1、smad2、smad3、COL1a2、JunB基因显著上调,和假手术组比较上调超过1.5倍,肾组织中的α-SMA、CD68表达面积和假手术组比较有显著统计学意义P<0.01。而RUUO组CHIP表达量较UUO组更是显著增强P<0.01,TGF-β1、smad2、smad3、COL1a2、JunB基因显著下调,和UUO组比较下调超过1.5倍,肾组织中的α-SMA、CD68表达面积减小和UUO组比较有显著统计学意义P<0.01。结论:发现CHIP的过量表达能显著降低细胞内,TGF-β1、Smad2/3、COL1a2、JunB基因水平,Western blot结果也表明CHIP蛋白表达能明显降低Smads介导的基因转录活性,进一步的基因芯片结果显示CHIP蛋白能明显下调TGF-β1所诱导的下游基因JunB的表达。结果提示CHIP蛋白能抑制性调节TGF-β1/Smads信号通路,抑制肾纤维化。

羊栖菜对UUO大鼠肾间质纤维化的影响

目的:观察羊栖菜对单侧输尿管梗阻(UUO)大鼠肾脏病变的影响。方法:建立UUO大鼠模型,用羊栖菜治疗3周,观察大鼠血肌酐、尿素氮、肾重、相对肾重,同时作病理学检查,并计算肾间质相对容量(Vv)。结果:UUO大鼠出现肾功能损害,病理显示出现明显的肾小管间质纤维化,羊栖菜治疗对肾功能减退有轻度抑制作用,能减少肾间质相对容量。结论:羊栖菜可能具有一定的抗肾间质纤维化作用。