虫草肾茶颗粒对单侧输尿管结扎大鼠肾间质纤维化模型CTGFmRNA的影响

目的观察虫草肾茶颗粒对单侧输尿管结扎致肾间质纤维化模型大鼠肾组织CTGFmRNA的影响。方法将60只雄性wistar大鼠随机分为五组,分别为假手术组、模型组、中药对照组、西药对照组、治疗组,每组12只。采用单侧输尿管结扎法制作肾间质纤维化模型,治疗组给予虫草肾茶为4ml.d-1.只-1;西药对照组给予福辛普利为1.5ml.d-1.只-1的水溶液;中药对照组给予肾衰宁胶囊4ml.d-1.只-1,1次/d给药;假手术组及模型组给予相同体积的生理盐水灌胃,1次/d。实验期间每周称一次体重,并观察大鼠生存状况。于第6周末处死大鼠,留取肾组织,应用realtime-PCR检测各组肾组织CTGFmRNA的表达。结果治疗组大鼠肾组织CTGFmRNA的表达与模型组相比明显下调。结论虫草肾茶颗粒可以部分抑制CTGF在单侧输尿管结扎大鼠肾组织的表达。

虫草肾茶颗粒对单侧输尿管结扎致肾间质纤维化大鼠肾脏影响的研究

目的观察虫草肾茶颗粒对单侧输尿管结扎致肾间质纤维化模型大鼠的肾脏保护作用,并探讨其机制。方法将60只雄性wistar大鼠随机分为五组,分别为假手术组、模型组、中药对照组、西药对照组、治疗组,每组12只。采用单侧输尿管结扎法制作肾间质纤维化模型,治疗组给予虫草肾茶为4ml·d-1·只-1;西药对照组给予福辛普利为1.5mg·d-1·只-1(浓度为0.375mg/ml)的水溶液;中药对照组给予肾衰宁胶囊4ml·d-1·只-1(相当于生药0.9g·ml·只-1),每日1次给药;假手术组及模型组给予相同体积的生理盐水灌胃,每日1次。实验期间每周称一次体重,并观察大鼠生存状况。分别于第2周、4周、6周采血,进行血肌酐(Scr)、尿素氯(BUN)检测。于第6周末处死大鼠,观察肾脏组织形态学变化,通过免疫组织化学方法检测各组大鼠肾组织结缔组织生长因子(CTGF)、核因子-κB(NF-κB)的表达。结果 (1)治疗组大鼠生存质量优于模型组;(2)治疗组大鼠各时间点BUN、Scr水平低于模型组,与中药对照组、西药对照组相比也有所降低;(3)治疗组大鼠肾组织CTGF、NF-κB的表达与模型组相比明显下调。结论虫草肾茶颗粒可通过减少CTGF、NF-κB在单侧输尿管结扎大鼠肾组织的表达,部分抑制肾间质纤维化的发生,保护肾脏,并能明显改善单侧输尿管结扎大鼠的生存质量,总体疗效优于肾衰宁及福辛普利。

含钆造影剂与肾源性纤维性皮肤病/肾源性系统性纤维化相关性展望

肾源性纤维性皮肤病/肾源性系统性纤维化(NFD/NSF)是一种在严重的肾病下自发获得的出疹性皮肤损害性疾病,可伴发全身皮肤和结缔组织系统性纤维化。这是一个慢性进展性和致死性疾病。其病因尚未明确。最近有报道指出使用含钆造影剂和NFD/NSF的发病有较强的相关性。本文就NFD/NSF的流行病学,临床病史和体格检查,病理生理学和组织学的特征,含钆造影剂在NFD/NSF发病中所扮演的角色,以及放射科医师在这个问题上应该注意的事项做一综述。

虫草益肾颗粒对5/6肾切除大鼠残肾组织α-平滑肌肌动蛋白表达的影响

目的观察虫草益肾颗粒对5/6肾切除大鼠残肾组织α-平滑肌肌动蛋白(α-SMA)与I型胶原表达的影响,探讨虫草益肾颗粒抗肾脏纤维化的作用机制。方法取雄性Wistar大鼠90只,采用改良的5/6肾切除法制作肾脏纤维化模型,将大鼠随机分为假手术组、模型组、百令胶囊对照组、虫草益肾颗粒低、高剂量组。低、高剂量组分别给予虫草益肾颗粒275mg.kg-1.d-1与550mg.kg-1.d-1灌胃,对照组给予百令胶囊315mg.kg-1.d-1灌胃,其余两组给予等量的蒸馏水灌胃,均为1次/d。实验期间检测大鼠24h尿蛋白定量的变化。连续给药12周后处死大鼠,观察肾脏组织形态学的变化,用半定量方法计算肾小球硬化指数与肾小管损伤指数。用免疫组织化学方法检测残肾组织α-SMA与I型胶原的表达。结果虫草益肾颗粒能减少5/6肾切除大鼠尿蛋白的排出(P<0.05,P<0.01),减轻肾小球硬化与肾小管损伤程度(P<0.05,P<0.01),下调残肾组织α-SMA与I型胶原的表达(P<0.05,P<0.01)。结论虫草益肾颗粒可能通过减少5/6肾切除大鼠尿蛋白的排出,抑制肾组织α-SMA与I型胶原的表达,从而抑制参与肾脏纤维化的细胞增殖与转分化,起到延缓肾脏纤维化进展的作用。

核因子-κB在慢性移植肾失功中作用机制的研究

目的探讨核因子-κB(NF-κB)及其下游信号分子调节激活正常T细胞表达和分泌的细胞因子(RANTES)和单核细胞趋化蛋白-1(MCP-1)在慢性移植肾失功(CRAD)患者移植肾组织的表达及与肾间质纤维化/小管萎缩(IF/TA)和尿蛋白的关系。方法采用免疫组化技术及计算机真彩色图像分析软件系统半定量检测103例病理诊断符合CRAD的移植肾组织NF-κB p65、RANTES及MCP-1表达量,并分析其表达与CRAD移植肾组织IF/TA、血肌酐及24h尿蛋白的关系。10例移植前零点穿刺正常肾组织作为对照组,入院行血清肌酐及24h尿蛋白定量检测。结果移植肾组织NF-κB p65表达较正常肾组织明显升高[(22.63±6.37)%、(38.59±5.36)%、(53.36±8.77)%比(7.83±0.57)%,P<0.01];其表达水平同炎症细胞趋化因子RANTES、MCP-1及IF/TA病理分级呈正相关(r分别为0.736、0.857和0.904,P均<0.01);NF-κB p65的表达与炎症细胞浸润亦具有正相关性(r=0.851,P<0.01);血肌酐及24h尿蛋白定量的变化随IF/TA病理分级增加而升高(r=0.902和0.870,P均<0.01)。结论在CRAD患者移植肾组织中,NF-κB p65表达明显增强,NF-κB p65及炎症趋化因子的高表达与移植肾间质炎症细胞浸润、肾间质纤维化及慢性移植肾失功等表现密切相关,提示它们可能参与了慢性移植肾功能不全的发生和发展。

ILK在非慢性排斥CAN患者移植肾组织的表达及其意义

目的探讨非慢性排斥CAN（慢性移植肾病）患者移植肾组织中整合素连接激酶（ILK）、转化生长因子β1（TGF-β1）的表达，及与移植肾间质纤维化／肾小管萎缩（IF／TA）的关系。方法用免疫组织化学技术和计算机真彩色图像分析系统半定量检测48例非慢性排斥CAN患者移植肾组织中ILK和TGF—β1的表达情况，分析二者间及与非慢性排斥CAN患者移植肾IF／TA病理分级之间的关系。15例正常肾组织作为对照。结果非慢性排斥CAN各组间的移植肾组织中ILK、TGF-β1的表达比正常肾组织明显增加（P〈0．01），并随IF／TA病理分级呈逐渐递增的趋势；非慢性排斥CAN移植肾组织中ILK的表达与TGF-β1呈正相关（r=0．930，P〈0．01）；ILK、TGF-β1表达水平分别与非慢性排斥CAN移植肾IF／TA病理分级呈正相关，（r=0．860、0．938；P〈0．01）；Scr与IF／TA病理分级呈正相关（r=0．790，P〈0．01）；ILK与SCr之间呈正相关关系（r=0．865，P〈0．001）。结论ILK可能是介导TGF-β1促进非慢性排斥CAN患者移植肾细胞外基质（extra cellular matrix，ECM）异常沉积发病机制，ILK在非慢性排斥CAN患者移植肾纤维化细胞信号通路中起重要作用。

Effects of Matrine on Adenine-Induced Chronic Tubulointerstitial Fibrosis in Rats

Aim ,To study the mechanism relative to therapeutic effects of matrine on adenine-induced renal interstitial fibrosis in rats. Methods Sixty male Wistar rats were selected and randomly divided into 3 groups： normal control group （NCG） consisted of 8 rats, adenine treated group （ATG） 28 rats, and matrine treated group （MTG） 24 rats. Each rat in ATG and MTG was gavaged with adenine （250 mg·kg^-1·d^-1 ） for 21 d. After gavage with adenine for one week, each rat in MTG was administered intraperitoneally matrine（20 mg·kg^-1·d^-1 ） in vehicle （ 1 mL of 0.9% sodium chloride） daily. On days 14, 21, and 28, the serum levels of urea nitrogen, creatinine, and IL-6 were determined and the rat kidneys of ATG, MTG and NCGwere examined pathologically. Radioimmunoassay for serum IL- 6 immunohistochemical staining for TGF-β1 expression in the kidney and semiquantitative analysis were performed. HE staining for semiquantitative analysis of tubulointerstitial injury. Results The serum levels of urea nitrogen and creatinine in MTG were lower as compare to ATG （ P 〈 0.05 ） whereas serum IL- 6 and renal TGF-β1 expression levels were significantly lower than those in ATG （P 〈0.05 ）, but all these indexes were higher than those in NCG （P 〈 0.01 ）. In MTG, the index of tubulointerstitial lesion was lower than that in ATG （P 〈 0.05 ）. Conclusion Matrine inhibits the renal tubulointerstial fibrosis in the adenine-induced rat model by suppressing serum level of IL-6 and expression of TGF-β1 in the tubulointerstitium.

TRANSFORMING GROWTH FACTOR-β1 AND SMAD4 SIGNALING PATHWAY DOWN-REGULATES RENAL EXTRACELLULAR MATRIX DEGRADATION IN DIABETIC RATS

Objective To investigate the role of transforming growth factor-131 （TGF-β1）/Smad4 pathway in development of renal fibrosis in streptozotocin （STZ）-induced diabetic nephropathy （DN） rats and explore its possible mechanism. Methods Male Wistar rats weighing 180-220 g were divided into 5 groups： group A （ normal control）, group B [ diabetes mellitus （DM） 2 weeks ], group C （ DM 4 weeks）, group D （ DM 8 weeks）, and group E （ DM 16 weeks）. Except for the normal control group, other groups were induced DM by single injection of STZ （55 mg/kg） respectively. Blood glucose level, serum creatinine, and 24-hour urine protein were examined. Expressions of TGF-β1 and Smad4 protein and mRNA in kidney were detected using immunohistochemical technique, Western blot, and real-time PCR. mRNA expressions of stromelysin-1 （ MMP-3 ）, tissue inhibitor of metalloproteinase-1 （ TIMP-1 ）, and collagen Ⅲ in kidney were also detected by real-time PCR. Results The levels of blood glucose, serum creatinine, and 24-hour urine protein in rats of group B, C, D, and E were higher than those of the control group. With the progression of renal fibrosis, the expressions of TGF-β1 and Smad4 protein and mRNA in kidney of diabetic rats elevated. In addition, the renal MMP-3 mRNA expression diminished in diabetic rats, while TIMP-1 and collagen Ⅲ mRNA increased. Conclusions In STZ-induced diabetic rats, the TGF-β1/Smad4 appears to play an important role in renal fibrosis of DN. The increased expression of TGF-β1 and Smad4 might result in the transcriptional regulation of downstream target genes of TGF-β1/Smad4 pathway, which contributes to the progression of renal fibrosis in diabetic rats.

Influence of ginsenoside Rg1, a panaxatriol saponin from Panax notoginseng, on renal fibrosis in rats with unilateral ureteral obstruction

Total saponins of Panax notoginseng （PNS） have been shown to ameliorate renal interstitial fibrosis. Ginsenoside Rg 1, a panaxatriol saponin, is one of the major active molecules from PNS. The present study was undertaken to investigate the effect of ginsenoside Rgl on renal fibrosis in rats with unilateral ureteral obstruction （UUO）. The rats were randomly divided into 3 groups： sham-operation （n=15）, UUO （n=15） and UUO with ginsenoside Rgl treatment （n=15, 50 mg per kg body weight, intraperitoneally （i.p.） injected）. The rats were sacrificed on Days 7 and 14 after the surgery. Histological examination demonstrated that ginsenoside Rgl significantly inhibited interstitial fibrosis including tubular injury as well as collagen deposition, u-smooth muscle actin （α-SMA） and E-cadherin are two markers of tubular epithelial-myofibroblast transition （TEMT）. Interestingly, ginsenoside Rgl notably decreased α-SMA expression and simultaneously enhanced E-cadherin expression. The messenger RNA （mRNA） of transforming growth factor-β1 （TGF-β1）, a key mediator to regulate TEMT, in the obstructed kidney increased dramatically, but was found to decrease significantly after administration of ginsenoside Rg 1. Further study showed that ginsenoside Rgl considerably decreased the levels of both active TGF-β1 and phosphorylated Smad2 （pSmad2）. Moreover, ginsenoside Rgl substantially suppressed the expression of thrombospondin-1 （TSP-1）, a cytokine which can promote the transcription of TGF-β1 mRNA and the activation of latent TGF-β1. These results suggest that ginsenoside Rgl inhibits renal interstitial fibrosis in rats with UUO. The mechanism might be partly related to the blocking of TEMT via suppressing the expression of TSP-1.

Expression of transforming growth factor beta-1 in fibrosis of transplanted kidney

Objective: In order to study the role and significance of transforming growth factor beta-1 (TGF-β1 )mRNA in transplanted renal fibrosis(TRF). Methods: Renal pathologic changes and expression of TGF-β1 mRNA were observed using in situ hybridization technique. The normal renal tissue as a. control group. Results: Expression of TGF-β1, mRNA in the renal fibrosis increased, compared with that in the control group. The expression rate were co-related to the stage of TRF. Conclusion: TGF-β1 is related to the pathogenesis and development of TRF.

Renin-angiotensin system in the pathogenesis of liver fibrosis

Hepatic fibrosis is considered a common response to many chronic hepatic injuries. It is a multifunctional process that involves several cell types, cytokines, chemokines and growth factors leading to a disruption of homeostatic mechanisms that maintain the liver ecosystem. In spite of many studies regarding the development of fibrosis, the understanding of the pathogenesis remains obscure. The hepatic tissue remodeling process is highly complex, resulting from the balance between collagen degradation and synthesis. Among the many mediators that take part in this process, the components of the Renin angiotensin system （RAS） have progressively assumed an important role. Angiotensin （Ang） II acts as a profibrotic mediator and Ang-（1-7）, the newly recognized RAS component, appears to exert a counter-regulatory role in liver tissue. We briefly review the liver fibrosis process and current aspects of the RAS. This review also aims to discuss some experimental evidence regarding the participation of RAS mediators in the pathogenesis of liver fibrosis, focusing on the putative role of the ACE2-Ang-（1-7）- Mas receptor axis.

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.

Research progress of renal interstitial fibrosis

Renal interstitial fibrosis （RIF） is a common pathological process of chronic kidney disease that progresses toend-stage renal failure. The degree of RIF is closely related to renal function. The study of the pathogenesis of renalinterstitial fibrosis, exploration of effective prevention measures to delay the progress of end stage renal disease andprolong the life of patients is significant. The pathology of RIF has complicated extracellular and intercellularmechanisms, involving many cells and cytokines, resulting in an incomplete mechanistic understanding of thedisease. Finding effective herbs or herbal extracts for prevention and treatment of RIF is crucial because currentmedical approaches do not reliably slow or reverse RIF. The research progress of RIF in recent years issummarized as follows.

The expression of AT1 receptor on hepatic stellate cells in rat fibrosis induced by CCl_4

Objectives To assess the effect of an ACE inhibitor and an Ang Ⅱ type 1 (AT1) receptor antagonist on preventing hepatic fibrosis induced by CCl4 in rats and to investigate whether there is the expression of AT1 receptors on hepatic stellate cells.Methods Studies were conducted in male Sprague-Dawley rats. Except for model group and control group, in three treated groups, either enalapril (5?mg/kg), or losartan (10?mg/kg), or enalapril+losartan were given to the fibrotic rats (daily gavage). Saline vehicle was given to the control group. After 6 weeks, liver fibrosis was assessed directly by hepatic morphometric analysis. The expression of AT1 receptors and α-smooth muscle actin (α-SMA) in liver tissue and isolated hepatic stellate cells (HSC) were detected by immunohistochemical techniques. Results Compared with the fibrosis in rats of the model group, rats treated with either enalapril or losartan, or a combination of two drugs, showed a limited expansion of the interstitium (P<0.05), but no significant difference was observed among the three treated groups (P>0.05). The expression of AT1 receptors was found in abundance in the fibrotic interstitium of the fibrotic rats, whereas in the normal control rats they were limited to the vascular wall. AT1 receptors were also expressed on activated HSC in culture plates. Conclusions Angiotensin-converting enzyme inhibitors and AT1 blockers might slow the progression of hepatic fibrosis. Activated HSCs expressed AT1 receptors. Activation of RAS might be related to hepatic fibrogenesis induced by CCl4.

The effect of L-arginine on the progression of chronic renal scarring in remnant kidney

OBJECTIVE: To investigate the effect of L-arginine (L-arg) on early compensatory renal growth (CRG), tubulointerstitial accumulation of extracellular matrix (ECM), long term survival rate and renal scarring in rats with 5/6 nephrectomy (SNx). METHODS: The experiment included four groups of rats (n = 5 each group): (1) Sham group, (2) SNx group, (3) SNx + L-arg group, and (4) Sham + L-arg group (L-arg 1% in drinking water). Parameters related with CRG and early tubulointerstitial expression of ECM and alpha smooth muscle actin (alphaSMA) were evaluated by immunohistochemistry at day 30. The survival rate and the extent of renal scarring in the rats were observed at day 120. RESULTS: L-arg significantly increased the early CRG of SNx rats as determined by the wet kidney weight (P

Sonic hedgehog signaling in kidney fibrosis: a master communicator

The hedgehog signaling cascade is an evolutionarily conserved pathway that regulates multiple aspects of embryonic development and plays a decisive role in tissue homeostasis. As the best studied member of three hedgehog ligands, sonic hedgehog(Shh) is known to be associated with kidney development and tissue repair after various insults. Recent studies uncover an intrinsic link between dysregulated Shh signaling and renal fibrogenesis. In various types of chronic kidney disease(CKD), Shh is upregulated specifically in renal tubular epithelium but targets interstitial fibroblasts, thereby mediating a dynamic epithelialmesenchymal communication(EMC). Tubule-derived Shh acts as a growth factor for interstitial fibroblasts and controls a hierarchy of fibrosis-related genes, which lead to the excessive deposition of extracellular matrix in renal interstitium. In this review, we recapitulate the principle of Shh signaling, its activation and regulation in a variety of kidney diseases. We also discuss the potential mechanisms by which Shh promotes renal fibrosis and assess the efficacy of blocking this signaling in preclinical settings. Continuing these lines of investigations will provide novel opportunities for designing effective therapies to improve CKD prognosis in patients.

Astragalus mongholicus ameliorates renal fibrosis by modulating HGF and TGF-β in rats with unilateral ureteral obstruction

Astragalus mongholicus （AM） derived from the dry root ofAstragalus membranaceus Bge. var. mongolicus （Bge.） Hsiao is a widely used traditional Chinese medicine. The present study investigated the potential role of AM on renal fibrosis on a rat model of unilateral ureteral obstruction （UUO）. We divided 48 Sprague-Dawley rats randomly into 4 groups： sham-operated group （Sham）, untreated UUO group, AM-treated （10 g/（kg.d）） UUO group, and losartan-treated （20 mg/（kg.d）） UUO group as positive control. Haematoxylin ＆ eosin （HE） and Masson staining were used to study the dynamic histological changes of the kidneys 7 and 14 d after operation. The expressions of fibronectin （FN）, type I collagen （coil）, hepatocyte growth factor （HGF）, transforming growth factor-β1 （TGF-β1）, and eL-smooth muscle actin （α-SMA） were analyzed by real-time polymerase chain reaction （PCR）, immunohistochemistry staining, and Western blot. Results show that, similar to losartan, AM alleviated the renal damage and decreased the deposition of FN and coil from UUO by reducing the expressions of TGF-β1 and α-SMA （P〈0.05）, whereas HGF increased greatly with AM treatment （P〈0.05）. Our findings reveal that AM could retard the progression of renal fibrosis. The renoprotective effect of AM might be related to inhibition ofmyofibroblast activation, inducing of HGF and reducing of TGF-β1 expression.

Distinct actions of intermittent and sustained β-adrenoceptor stimulation on cardiac remodeling

Heart disease is associated with increased sympathetic nerve activity and elevated levels of circulating catecholamines,resulting in chronic stimulation of the β-adrenergic receptors (β-AR) and consequent pathological cardiac remodeling.Experimentally,chronic administration of the β-AR agonist isoproterenol (ISO) has been most commonly used to model β-AR-induced cardiac remodeling.However,it remains unclear whether β-AR-mediated cardiac remodeling and dysfunction differs between sustained versus pulsatile (intermittent) exposure to a β-agonist.Here,we compare the effects of intermittent versus sustained administration of ISO on cardiac remodeling and function in mice.Animals were administered 5 mg (kg d)-1 ISO for 2 weeks either by daily subcutaneous injection,or continuous infusion via an implanted osmotic minipump.Cardiac function and remodeling were determined by echocardiography,micromanometry and histology.Moreover,Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) were utilized to define the proteins and genes involved.Both sustained and intermittent administration of ISO resulted in a similar degree of cardiac hypertrophy (16% and 19%,respectively).However,mice receiving ISO by daily injection developed more severe ventricular systolic and diastolic dysfunction and myocardial fibrosis compared with mice receiving ISO via the osmotic minipump.The disparity in results between the delivery methods is suggested to be due,at least in part,to increased expression of fibrogenic factors,including connective tissue growth factor (CTGF) and NADPH oxidase (NOX4),in mice receiving intermittent application of ISO.In summary,compared with sustained exposure to a β-AR agonist,intermittent β-AR stimulation leads to more severe cardiac dysfunction and fibrosis.These findings not only further our understanding of β-AR function in the setting of cardiac pathophysiology,but also highlight that significant differences can result dependent upon the mode of experimental β-AR stimulation in inducing cardiomyopathy.

Inhibition mechanism of compound ethanol extracts from Wuweizi(Fructus Schisandrae Chinensis) on renal interstitial fibrosis in diabetic nephropathy model mice

OBJECTIVE：To evaluate inhibition effect and mech- anism of compound ethanol extracts from Wuweizi （Fructus Schisandrae Chinensis）, Chuanxiong （Rhi- zoma Chuanxiong） and Muli （Cocha Ostreae） （FRC） on glomerular and tubular interstitial fibrosis in streptozocin （STZ）-induced diabetic nephropathy （ND） model mice. METHODS： Twenty-seven male C57BL/6 mice were divided randomly into 3 groups： nondibetic （ND）, STZ-induced diabetic （D）, and STZ-induced diabetic that were treated with .5 g. kg1. daylof FRC by oral gavage （DFRc）, with 9 in each group. The protein ex- pressions of E-cadherin, a-smooth muscle actin （a-SMA）, Plasminogen Activator Inhibitor-1 （PAl-l） in renal tissues were investigated by Western blot- ting. The expressions of fibronectin （FN） and o-SMA were detected by immunohistochemical method. The morphological changes of renal tissues were observed under a microscope. RESULTS： Renal tissues in the DFRC group showed a lessened degree of fibrosis. Meanwhile, the expres- sions of FN, o-SMA and PAl-lwere significantly lower in the DrRc group than those in the D group （all P〈0.05）.CONCLUSION： FRC can ameliorate the DN in the C57BL/6 mice, and its mechanism may relate to in- hibition on the epithelial to mesenchymal transdif- ferentiation, endothelial-myofibroblast transition and PAl-1 expression.

Effect of Tangnaikang on TGF-β_1-induced transdifferentiation of human renal tubular epithelial HK-2 cells

OBJECTIVE： To explore the function of Tangnai- kang （TNK） in the prevention and treatment of re- nal interstitial fibrosis through transdifferentiation of the human renal tubular epithelial cell line HK-2 induced bytransforming growth factor-β1 （TGF-β1）. METHODS： HK-2 cells cultured in dulbecco＇s modi- fied eagle medium/F12 （1：1） with 10% fetal calf se- rum were divided into six groups： blank control group, TGF-β1 group （TGF-β1 10 ng/mL）, serum con- trol group （TGF-β1 10 ng/mL ＋ 10% serum）, treat- ment group 1 （TGF-β1 10 ng/mL ＋ 5% TNK serum）, treatment group 2 （TGF-β1 10 ng/mL＋10% TNK se-rum）, and treatment group 3 （TGF-β1 10 ng/mL＋ 20% TNK serum）. Cell proliferation was detected by 4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazoliu m bromide assay. Expression of a-smooth muscle ac- tin （a-SMA） and E-cadherin were observed by im- munohistochemical assay. The contents of collagen Ⅰ （Col Ⅰ）, collagen Ⅲ（ColⅢ）, and fibronectin （FN） in the culture medium supernatant were detected by ELISA. RESULTS： E-cadherin was expressed and α-SMA was not expressed in normal HK-2 cells. In HK-2 cells cultured with TGF-β1, α-SMA expression signifi- cantly increased, HK-2 cells significantly proliferat- ed, and secretion of Col Ⅰ, Col Ⅲ, and FN significantly increased compared with the blank control group （all P〈0.05）. In the HK-2 cells cultured with TGF-β1 and TNK serum, the expression of α-SMA signifi- cantly decreased, the expression of E-cadherin sig- nificantly increased, and the cell proliferation and the secretion of Col Ⅰ, Col Ⅲ and FN were significant- ly inhibited compared with the TGF-β1 group （all P〈 0.05. CONCLUSION： TNK can inhibit cell proliferation and reduce secretion of Col Ⅰ, Col Ⅲ, and FN.This in- dicates that TNK can inhibit transdifferentiation of human renal tubular epithelial cells induced by TGF-β1, with the effect of preventing and treating renal interstitial fibrosis.