黄芪多糖对早期糖尿病肾病大鼠足细胞nephrin和podocin表达的影响

目的: 观察黄芪多糖(APS)对糖尿病肾病(DN)大鼠足细胞裂孔隔膜蛋白分子(nephrin和podocin)表达的影响,探讨其防治DN的作用机制。方法: 24只健康雄性SD大鼠,随机抽取8只为正常对照组,其余大鼠链脲佐菌素(STZ)腹腔注射复制糖尿病模型,1周后经血糖浓度测量确定大鼠糖尿病建模成功,将造模大鼠分为STZ组(8只)和STZ+APS组(8只),各组分别干预8周。于干预后第2、5、8周检测大鼠血糖浓度;第8周末称量大鼠体重,计算肾指数,观察肾脏病理改变;检测24 h 尿蛋白总量t和 血尿素氮、血肌酐;蛋白免疫印迹法检测大鼠肾组织nephrin和podocin的表达水平。结果: 应用APS干预后,大鼠血糖、肾系数、24 h尿蛋白总量、血尿素氮和血肌酐明显降低(P<0.05),体重增加(P<0.05);病理改变减轻;nephrin和podocin蛋白表达增加(P<0.05)。结论: APS对DN肾脏的保护作用可能与维持足细胞nephrin和podocin的表达有关。     

真武汤对链脲佐菌素所致大鼠糖尿病肾病的保护作用

 目的：探讨真武汤对糖尿病大鼠肾脏的保护作用及机制。方法：腹腔注射链脲佐菌素(STZ)建立糖尿病肾病大鼠模型,将成模大鼠随机分为糖尿病肾病(DN)模型组和真武汤治疗组（真武组）,另设正常组。采用生化、HE染色观察真武汤对糖尿病肾病大鼠肾功能、肾组织形态学变化及脂质过氧化相关参数的作用;采用蛋白免疫印迹方法探讨真武汤对肾组织α-平滑肌肌动蛋白（α-SMA）和NF-κB表达的影响。结果：模型组大鼠肾系数、24 h尿蛋白定量、血尿素氮、肌酐、血糖和丙二醛(MDA)均显著升高(P<0.05),体重、超氧化物歧化酶(SOD)及诱导型一氧化氮合酶（iNOS）显著降低(P<0.05);真武组大鼠的肾系数、24 h尿蛋白、尿素氮、肌酐、血糖和MDA明显低于模型组(P<0.05),iNOS显著高于模型组（P<0.05）;模型组大鼠肾小球肥大、毛细血管基底膜增厚,系膜基质增生,肾小管上皮细胞空泡样变,可见蛋白管型;真武组病变轻于模型组。模型大鼠肾组织α-SMA及NF-κB蛋白的水平明显高于正常组（P<0.05）,真武组大鼠肾组织α-SMA及NF-κB蛋白水平明显低于模型组（P<0.05）。结论：真武汤能减轻糖尿病肾病肾脏局部氧化应激反应,改善糖尿病肾病大鼠肾功能,减轻病理损伤,其发挥肾脏保护作用可能与抑制α-SMA及NF-κB蛋白的表达有关。     

VEGF受体抑制剂对Ⅰ型糖尿病肾病大鼠足细胞病的治疗作用

目的:探讨血管内皮生长因子(VEGF)受体抑制剂SU5416对糖尿病肾病足细胞病的治疗作用.方法:SPF级SD雄性大鼠共30只,设为正常对照组(NC)、糖尿病肾病模型组(DN)、SU5416治疗组(SU5416),每组10只.DN大鼠由链脲菌素(STZ)诱导形成.SU5416治疗后第8周,分别测定大鼠体重( body weight,BW)、肾重(kidney weight,KW)、血糖( glucose,Glu)、大鼠24h尿蛋白排泄率(24 h UAER)和血肌酐(Scr).免疫荧光技术观察肾脏足细胞标记蛋白nephrin和podocin表达,Real time-PCR技术检测nephrin、podocin和VEGF mRNA水平,并观察肾脏病理变化.结果:与NC组大鼠相比,DN组大鼠BW明显下降,KW增加显著(P＜0.05),24 h UAER、Glu和Scr均明显升高,肾组织VEGF mRNA水平明显升高,nephrin、podocin蛋白水平及mRNA水平明显下降(P＜0.05),肾小球基底膜增厚、系膜基质增生;SU5416治疗后大鼠肾脏病理改善,KW较DN组明显下降,24hUAER明显降低,nephrin、podocin蛋白水平及mRNA水平上调(P＜0.05),但BW、Glu、Scr和VEGF mRNA水平治疗前后无统计学差异(P＞0.05).结论:SU5416治疗能降低DN大鼠24 h UAER、改善肾脏病理表现和上调肾小球足细胞标记蛋白nephrin和podocin水平.表明VEGF受体抑制剂对DN的治疗作用与其足细胞保护有关,抑制VEGF活性可能成为控制DN足细胞病进展的治疗手段之一.     

过表达APMAP减轻阿霉素肾病肾小球足细胞损伤

目的 探讨脂肪细胞膜相关蛋白质(APMAP)过表达对阿霉素(ADR)肾病肾小球足细胞损伤的影响。方法 采用尾静脉注射ADR构建阿霉素肾病大鼠模型，免疫组化观察肾组织中APMAP、NF-κB p65蛋白表达情况。构建APMAP基因过表达的鼠源肾小球足细胞MPC-5细胞株，并以0.5μmol/L ADR体外诱导构建足细胞损伤模型，再联合NF-κB信号通路激活剂CU-T12-9进行处理。CCK-8检测细胞增殖活性；ELISA检测乳酸脱氢酶(LDH)活性；流式细胞术检测细胞凋亡率；Western blot检测NF-κB p65、p-NF-κB p65、TNF-α等蛋白表达。结果 APMAP在阿霉素肾病大鼠肾组织中低表达，而NF-κB p65高表达(P<0.05)。APMAP过表达可提高ADR暴露下MPC-5细胞增殖活性，降低LDH活性及细胞凋亡率，下调NF-κB p65、p-NF-κB p65、TNF-α等蛋白表达(P<0.05);联合CU-T12-9处理可显著抑制APMAP过表达对ADR暴露下MPC-5细胞损伤的改善作用。结论 过表达APMAP可抑制ADR诱导的肾小球足细胞损伤，...     

细胞凋亡及其相关基因蛋白在阿霉素肾病中的作用

目的：探讨细胞凋亡及其相关基因蛋白在阿霉素（ADR）肾病发病中的作用及机制。 方法：复制ADR肾病模型，用超氧化物歧化酶（SOD）干预，用原位末端标记法、免疫组化法、原位杂交法分别检测细胞凋亡、基因蛋白表达、野生型p53 mRNA，同时测定肾皮质匀浆丙二醛（MDA）及谷胱甘肽过氧化物酶（GSH-Px）水平。结果：模型组皮质部肾小球、近曲及远曲肾小管细胞凋亡数和c-Myc蛋白及野生型p53转录水平、近曲和远曲肾小管细胞Bax蛋白表达、肾皮质MDA水平明显高于对照组（P<0.01），肾皮质GSH-Px活性、近曲和远曲肾小管细胞Bcl-2蛋白表达明显低于对照组（P<0.01）；SOD组肾小球和近曲、远曲肾小管细胞凋亡数明显少于模型组（P<0.01）。结论：ADR肾病病鼠的发病与肾小球、近曲和远曲肾小管细胞凋亡有关，氧自由基（OFR）可能通过上调诱导细胞凋亡的基因蛋白表达，下调抑制细胞凋亡的基因蛋白表达而引起细胞凋亡。     

白花丹参水提取物通过调控NOD2基因表达对高糖诱导小鼠肾脏足细胞损伤的保护作用及其机制研究

目的研究白花丹参水提取物对高糖诱导的小鼠肾脏足细胞MPC5损伤的影响及分子作用机制。方法高糖刺激和白花丹参水提取物处理MPC5细胞,qRT-PCR检测NOD2、podocin和nephrin mRNA表达,Western blot检测NOD2、podocin和nephrin蛋白表达,流式细胞术测定细胞凋亡率。结果 NOD2在大多数人糖尿病肾病组织中呈高表达;高糖抑制小鼠足细胞MPC5中podocin和nephrin的表达,促进MPC5细胞凋亡,诱导MPC5细胞中NOD2 m RNA和蛋白的表达;白花丹参水提取物可以促进高糖诱导的MPC5细胞中podocin和nephrin的表达,抑制NOD2基因的表达,抑制高糖诱导的MPC5细胞凋亡,抑制NOD2表达可促进高糖诱导的MPC5细胞中podocin和nephrin的表达,并抑制细胞凋亡;过表达NOD2逆转了白花丹参水提取物对高糖诱导的MPC5细胞损伤的作用。结论白花丹参水提取物可能通过调控NOD2基因表达减轻高糖诱导的小鼠肾脏足细胞MPC5的损伤,抑制细胞凋亡。白花丹参水提取物对糖尿病肾病具有潜在治疗作用。     

微胶囊化儿茶素对阿霉素肾病大鼠VEGF表达的影响

目的： 研究微胶囊化儿茶素对阿霉素肾病大鼠血管内皮生长因子(VEGF)表达的影响。方法: 将120只雌性SD大鼠随机分为对照组、肾病组、地塞米松组、维生素E组、儿茶素组和微胶囊组共6组,尾静脉1次性注射阿霉素（5 mg/kg BW）制备肾病模型;利用酶联免疫吸附试验测定血清与尿中VEGF含量;利用免疫组织化学方法检测肾组织中VEGF表达。结果: 实验第4周末（维生素E组除外）与第6周末,各组大鼠尿、血清及肾组织中VEGF含量均显著高于对照大鼠、显著低于肾病大鼠（均P<0.01）,实验末，微胶囊组大鼠尿与血清中VEGF含量明显低于儿茶素组（分别P<0.01，P<0.05）,微囊组大鼠肾组织VEGF表达低于儿茶素组,但无显著差异。微囊化儿茶素治疗组24 h尿蛋白排泄量显著低于儿茶素治疗组（P<0.05）,24 h尿蛋白排泄与尿、血清及肾组织中VEGF含量均显著正相关（P<0.01）。结论: 儿茶素降低阿霉素肾病大鼠尿蛋白排泄可能是通过降低VEGF的排泄与分泌实现的，儿茶素微囊化后，有助于提高其降低VEGF的排泄与分泌，减少尿蛋白的排泄。     

糖尿病大鼠肾脏nephrin表达的动态观察

目的从不同时间点动态观察足细胞相关分子nephrin在糖尿病大鼠肾组织中的表达,探讨nephrin在DN蛋白尿发生发展中的改变及意义。方法将SD大鼠分为健康对照组(C组)、糖尿病组(DM组)。SD大鼠腹腔注射链脲佐菌素制成糖尿病模型,正常对照组注射等量枸橼酸缓冲液。分别于0、2、4、6、8、12周动态检测尿白蛋白(UA),血糖(BG)及肾功能,取肾组织行常规病理检查,免疫组化、Western blot分析技术检测肾脏nephrin的表达。结果 (1)与NC组相比,DM组随着病程的延长,尿白蛋白、BUN、Ccr均逐渐出现明显的变化,而白蛋白尿早于BUN、Ccr异常的出现;(2)与NC组相比,DM组足细胞相关分子nephrin蛋白于第2周出现表达下调,随着时间的进展,nephrin蛋白的表达进一步减少,直至第12周,各周间比较差异有显著性(P0.05);(3)相关分析显示:尿白蛋白排泄与足细胞相关分子nephrin呈负相关。结论糖尿病大鼠早期(2周)出现足细胞相关分子nephrin表达下调,是糖尿病肾病早期损伤指标,nephrin参与了糖尿病肾病大鼠蛋白尿的发生及发展。     

造影剂肾病发病机制中炎症反应的探讨*

  目的：通过观察肿瘤坏死因子α（TNF-α）与核因子κB（NF-κB）在造影剂肾病(CIN)模型大鼠肾组织中的表达,初步探讨CIN发病中是否存在炎症反应机制。方法：将96只雄性SD大鼠随机分成2组：模型组（n=48）和对照组（n=48）,分别从尾静脉注射碘造影剂和生理盐水10 mL/kg。在注射后6 h、12 h、24 h、48 h、72 h、5 d、10 d和15 d各处死6只大鼠,留取血液和肾组织,采用HE染色法观察肾脏病理变化,免疫组化和RT-PCR法分别检测肾损伤分子1（KIM-1）、NF-κB、TNF-α蛋白和mRNA表达情况,并进行相关性分析。结果：（1）对照组血清肌酐（SCr）和血尿素氮（BUN）各时点变化不大（P＞0.05）,模型组各时点（除15 d外）SCr和BUN水平明显高于对照组（P＜0.05）;（2）对照组各时点肾小管无明显损伤,病理评分无显著差异（P＞0.05）。模型组的肾小管损伤评分显著高于同一时点的对照组（P＜0.05）;（3）各因子在造膜后6 h后开始大量表达,KIM-1蛋白及mRNA在24 h达高峰,NF-κB、TNF-α蛋白及mRNA在48 h达高峰,且与对照组对应时点（除15 d外）比较均有显著差异（P＜0.05）;（4）模型组肾小管损伤评分与NF-κB、TNF-α蛋白及mRNA表达呈正相关（r=0.843、0.758、0.743和0.707,P＜0.05）;模型组肾组织的NF-κB、TNF-α蛋白及mRNA表达与KIM-1蛋白及mRNA表达呈正相关（r=0.863、0.807、0.839和0.855,均P＜0.05）。结论：NF-κB和TNF-α在CIN大鼠肾脏中的表达上调,其表达水平与肾小管损伤程度相关。CIN的发生发展中存在炎症反应机制。     

VEGF受体抑制剂对I型糖尿病肾病大鼠足细胞病的治疗作用

目的:探讨血管内皮生长因子(VEGF)受体抑制剂SU5416对糖尿病肾病足细胞病的治疗作用。方法:SPF级SD雄性大鼠共30只,设为正常对照组(NC)、糖尿病肾病模型组(DN)、SU5416治疗组(SU5416),每组10只。DN大鼠由链脲菌素(STZ)诱导形成。SU5416治疗后第8周,分别测定大鼠体重(body we ight,BW)、肾重(k idney we ight,KW)、血糖(glucose,G lu)、大鼠24 h尿蛋白排泄率(24 h UAER)和血肌酐(Scr)。免疫荧光技术观察肾脏足细胞标记蛋白nephrin和podoc in表达,Real tim e-PCR技术检测nephrin、podoc in和VEGF mRNA水平,并观察肾脏病理变化。结果:与NC组大鼠相比,DN组大鼠BW明显下降,KW增加显著(P<0.05),24 h UAER、G lu和Scr均明显升高,肾组织VEGF mRNA水平明显升高,nephrin、podoc in蛋白水平及mRNA水平明显下降(P<0.05),肾小球基底膜增厚、系膜基质增生;SU5416治疗后大鼠肾脏病理改善,KW较DN组明显下降,24 hUAER明显降低,nephrin、podoc in蛋白水平及mRNA水平上调(P<0.05),但BW、G lu、Scr和VEGF mRNA水平治疗前后无统计学差异(P>0.05)。结论:SU5416治疗能降低DN大鼠24 h UAER、改善肾脏病理表现和上调肾小球足细胞标记蛋白nephrin和podoc in水平。表明VEGF受体抑制剂对DN的治疗作用与其足细胞保护有关,抑制VEGF活性可能成为控制DN足细胞病进展的治疗手段之一。     

Direct effect of plasma permeability factors from patients with idiopatic FSGS on nephrin and podocin expression in human podocytes

The presence of circulating plasma factors (PF) altering renal permeability to proteins has been previously described in patients with focal segmental glomerulosclerosis (FSGS). Since these patients show reduced nephrin and podocin expression at renal biopsy, we evaluated the effect of serum and PF from patients with FSGS on nephrin and podocin expression in human podocytes. We studied 7 sera from patients with steroid-resistant FSGS, 3 from patients with nephrotic syndrome caused by non-immune disease, and 6 from healthy subjects. PF was prepared from plasmapheresis eluates of 2 patients with post-transplant recurrence of FSGS. Purification procedure was based on protein A Sepharose chromatography and differential precipitation in ammonium sulphate. Nephrin and podocin expression was semi-quantitatively evaluated by immunofluorescence. We found that serum and PF from FSGS patients rapidly induced redistribution and loss of nephrin in podocytes. This effect was associated with cytoskeleton redistribution and inhibited by cytochalasin B and sodium azide. On the contrary, podocin expression was unchanged after incubation with serum and PF from FSGS patients for short periods, but markedly reduced at 24 h. Our results demonstrate that serum and PF from FSGS patients may directly affect nephrin and podocin in human podocytes, thus providing new insights into the mechanisms causing proteinuria in FSGS.     

Mesenchymal Stem Cells Ameliorate Podocyte Injury and Proteinuria in a Type 1 Diabetic Nephropathy Rat Model

Mesenchymal stem cells (MSC) attenuate albuminuria and preserve normal renal histology in diabetic mice. However, the effects of MSC on glomerular podocyte injury remain uncertain. The aim of this study was to evaluate the effects of MSC on podocyte injury in streptozotocin (STZ)-induced diabetic rats. Thirty days after diabetes induction by STZ injection (65 mg/kg, intraperitoneally) in Sprague-Dawley rats, the diabetic rats received medium or 2 × 10⁶ enhanced green fluorescent protein-labeled MSC via the renal artery. In vivo tracking of MSC was followed by immunofluorescence analysis. Diabetes-related physical and biochemical parameters were measured on day 60 after the MSC infusion. The expression of podocyte markers (nephrin and podocin), podocyte survival factors (VEGF and BMP-7), and the ultrastructural pathology of podocytes were also assessed. MSC were only detected in the glomeruli from the left kidney receiving MSC infusion. Compared with medium-treated diabetic rats, rats treated with MSC showed a suppressed increase in kidney weight, kidney to body weight index, creatinine clearance rate, and urinary albumin to creatinine ratio; however, the treatment had no effect on blood glucose or body weight levels. Furthermore, the MSC treatment reduced the loss of podocytes, effacement of foot processes, widening of foot processes, thickening of glomerular basal membrane (GBM), and loss of glomerular nephrin and podocin. Most important, MSC-injected kidneys expressed higher levels of BMP-7 but not of VEGF. Our results clearly demonstrated that intra-arterial administration of MSC prevented the development of albuminuria as well as any damage to or loss of podocytes, though there was no improvement in blood sugar levels. The protective effects of MSC may be mediated in part by increasing BMP-7 secretion.     

Molecular basis of the functional podocin-nephrin complex: mutations in the NPHS2 gene disrupt nephrin targeting to lipid raft microdomains

Hereditary nephrotic syndrome is a heterogeneous disease, characterizedby heavy proteinuria and renal failure. Mutations of NPHS1 orNPHS2, the genes encoding for nephrin and podocin, lead to earlyonset of heavy proteinuria, and rapid progression to end-stagerenal disease, suggesting that both proteins are essential forthe integrity of the glomerular filter. Podocin is a stomatinprotein family member with a predicted hairpin-like structurelocalizing to the insertion site of the slit diaphragm of podocytes,the visceral glomerular epithelial cells of the kidney. Herewe investigate the pathomechanisms of different disease-causingpodocin mutations. We show that wild-type podocin is targetedto the plasma membrane, and forms homo-oligomers involving thecarboxy and amino terminal cytoplasmic domains. The associationof podocin with specialized lipid raft microdomains of the plasmamembrane was a prerequisite for recruitment of nephrin intorafts. In contrast, disease-causing mutations of podocin (R138Qand R138X) failed to recruit nephrin into rafts either becausethese mutants were retained in the endoplasmic reticulum (R138Q),or because they failed to associate with rafts (R138X) despitetheir presence in the plasma membrane. None of the mutants didaugment nephrin signaling, suggesting that lipid raft targetingfacilitates nephrin signaling. Our findings demonstrate thatthe failure of mutant podocin to recruit nephrin into lipidrafts may be essential for the pathogenesis of NPHS2. ^* To whom correspondence should be addressed. Tel: +49 7612703559;Fax: +49 7612706362; Email: benzing{at}med1.ukl.uni-freiburg.de      

Effect of Shen-qi-di-huang decoction on reducing proteinuria by preserving nephrin in adriamycin-induced nephropathy rats

The aim of this study is to investigate the effect of Shen-qi-di-huang decoction on reducing proteinuria and to discuss the mechanism of its action in Adriamycin (ADR)-induced nephropathy rats. The rats were randomly divided into three groups (n=12 each group): normal control (group A); ADR model control (group B); ADR + Shen-qi-di-huang decoction (group C). In group B and C, the rats were intravenously injected with ADR (6.5mg/kg). The rats in group C were orally administrated with Shen-qi-di-huang decoction after the injection of ADR. On day 7, 14, 28, 56 after ADR injection, 24h urine protein was detected. On day 28, 56 after ADR injection, ALB, ALT, serum creatinine (Scr) and BUN were examined. The morphological changes of the kidneys were observed by light microscope and electron microscope on day 28, 56 after ADR injection. The expression of nephrin was determined by immunohistochemistry and RT-PCR on day 28, 56 after ADR injection. Compared with group B, 24h urine protein and Scr decreased in group C on day 56 (P<0.05). The expression of nephrin determined by immunohistochemistry and RT-PCR increased in group C on day 28, 56 (P<0.05). The morphology observed by light microscope and electron microscope improved in group C on day 28, 56. Shen-qi-di-huang decoction decreases proteinuria, protects kidney function, and ameliorates histopathology in ADR-induced rats by preserving nephrin expression.     

Key molecular events in puromycin aminonucleoside nephrosis rats

Nephrin, podocin and alpha-actinin are all involved in proteinuria, but it is unclear which molecular event plays a crucial role during the development of proteinuria. Immunofluorescence staining and real-time quantitative polymerase chain reaction were used to study the glomerular expression of these molecules in puromycin aminonucleoside (PAN) nephrosis. Morphometric methods were applied to evaluate the podocyte foot process (FP) morphology. Two days after PAN injection, nephrin and podocin staining became discontinuous, podocin intensity decreased and FP swelled. Nephrin protein and mRNA decreased at day 5. Both podocin and nephrin intensity decreased dramatically when heavy proteinuria occurred, but nephrin mRNA was regained. When proteinuria disappeared, podocin recovered whereas nephrin did not (P = 0.02); alpha-actinin intensity increased (P = 0.009) and the distribution changed. The podocyte FP volume density correlated negatively with nephrin (r = -0.78, P = 0.0001) and podocin immunofluorescence intensity (r = -0.76, P = 0.0001). We conclude that, before the onset of proteinuria, the first response was the nephrin and podocin distribution change, podocin protein decrease and swollen FP; the podocin quantitative change was earlier than nephrin. Podocin and nephrin distribution and the protein level was associated with proteinuria more closely than their mRNA level. The delayed alpha-actinin induction might be a reparative response.     

厄贝沙坦对早期糖尿病肾病大鼠足细胞nephrin表达的影响

目的： 观察nephrin在糖尿病肾病（DN）大鼠肾小球足细胞中的表达，探讨厄贝沙坦对DN肾脏保护作用的机制。方法： 应用链脲佐菌素建立大鼠DN模型，将DN模型大鼠随机分为2组：厄贝沙坦治疗组、模型对照组；另设正常对照组。各组分别干预8周后，观察大鼠体重、肾重、相对肾重、24 h尿蛋白定量、血糖、血清尿素氮、肌酐、总胆固醇、甘油三酯变化，利用光镜、电镜观察肾脏病理改变，应用免疫组化技术观察nephrin表达情况。结果： 应用厄贝沙坦干预后，DN大鼠24 h尿蛋白定量明显减少、肾功能改善、肾脏病理改变显著减轻、足细胞nephrin表达量明显减少。结论： 厄贝沙坦对DN肾脏的保护作用可能与其抑制足细胞nephrin表达有关。     

沉默微小RNA-218表达保护STZ诱导的糖尿病大鼠肾组织

目的:探讨沉默微小RNA-218(microRNA-218,miR-218)表达对链脲佐菌素(streptozotocin,STZ)诱导的糖尿病肾病大鼠肾脏组织的保护作用及其可能机制。方法:采用单次腹腔注射STZ(50 mg/kg)方法制备糖尿病大鼠模型并构建miR-218短发夹RNA(short hairpin RNA,shRNA)慢病毒载体。SD大鼠被随机分为健康对照组、糖尿病模型组、空载慢病毒组及miR-218-shRNA组。于自动生化仪上检测不同时点(4、8和12周)大鼠血糖、24h尿蛋白量、血清肌酐(serum creatinine,SCr)及血尿素氮(blood urea nitrogen,BUN)含量。实时荧光定量PCR(RTqPCR)检测肾脏组织miR-218的表达。RT-qPCR和Western blot检测血红素氧合酶1(heme oxygenase-1,HO-1)、肾病蛋白(nephrin)和p38丝裂原激活的蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)的mRNA及蛋白表达水平。Caspase-3活性检测试剂盒检测caspase-3活性。末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling,TUNEL)法检测肾脏组织细胞凋亡。结果:与健康对照组相比,STZ处理后大鼠miR-218表达水平显著升高。同时模型大鼠的血糖、24 h尿蛋白量、SCr及BUN含量显著升高(P0.05);模型大鼠肾脏组织中HO-1和nephrin的mRNA和蛋白表达水平显著降低,而p38 MAPK蛋白的磷酸化水平显著升高;另外,模型大鼠肾脏组织中的caspase-3活性也显著升高。模型大鼠感染miR-218-shRNA后,miR-218表达水平显著下降并可以显著逆转上述效应。miR-218-shRNA组肾脏组织细胞的凋亡水平显著低于糖尿病模型组及空载慢病毒组。结论:miR-218参与了糖尿病大鼠的肾脏损伤,慢病毒载体沉默其表达能有效抑制肾脏组织细胞的凋亡,提示miR-218可以作为糖尿病肾病的基因治疗靶点。