雷公藤多苷对阿霉素肾病大鼠Dicer酶的影响

雷公藤制剂被用于治疗肾小球肾炎已有30多年的历史,在减少尿蛋白方面,其疗效显著[1].目前已发现其抗蛋白尿作用与恢复nephrin、podocin表达有关.我们用阿霉素肾病大鼠模型进行研究,进一步探讨其抗蛋白尿作用机制.     

雷公藤多苷对阿霉素肾病大鼠Dicer酶的影响

雷公藤制剂被用于治疗肾小球肾炎已有30多年的历史,在减少尿蛋白方面,其疗效显著[1].目前已发现其抗蛋白尿作用与恢复nephrin、podocin表达有关.我们用阿霉素肾病大鼠模型进行研究,进一步探讨其抗蛋白尿作用机制.     

雷公藤多苷对糖尿病肾病大鼠足细胞病变的影响

目的 探讨雷公藤多苷（TWP）对糖尿病肾病大鼠足细胞病变的影响。 方法 SD大鼠100只,按随机数字表法分为正常对照组（A组）、糖尿病组（B组）与TWP治疗组（C组）。并将C组按4、8、16 mg?kg-1?d-1不同给药剂量分为3组（Ca、Cb、Cc）。糖尿病组与TWP治疗组大鼠分别给予链脲菌素（STZ）一次性腹腔注射建立糖尿病大鼠模型。12周后检测24 h尿蛋白量、血肌酐（Scr）、尿素氮（BUN）、外周血白细胞（WBC）、血糖（Glu）、肾质量/体质量（KW/BW）的变化;HE染色检测肾组织病理变化;透射电镜测量足细胞超微结构变化;免疫荧光法检测肾皮质nephrin和podocin蛋白表达。 结果 （1）与A组比较,B组及C组Scr、BUN增高（P ＜ 0.05）;Glu、KW/BW和24 h尿蛋白量显著增高（P ＜ 0.01）,除Cc组（3/20）出现肝酶（ALT、AST）增高及WBC下降外,各组ALT、AST、WBC差异无统计学意义;肾脏皮质nephrin和podocin蛋白表达减少,肾小球、小管间质及足细胞病变明显。（2）与B组比较,C组KW/BW和24 h尿蛋白量降低（P ＜ 0.01）,肾脏皮质nephrin和 podocin蛋白表达增高（均P ＜ 0.01）,肾小球、小管间质及足细胞病变明显减轻,并随雷公藤多苷剂量的增加而越加明显。 结论 TWP对糖尿病大鼠足细胞病变具有保护作用,并与剂量相关。其部分机制可能与上调nephrin和podocin蛋白表达有关。     

雷公藤多苷对糖尿病肾病大鼠肾脏中CTGF表达的影响

目的：观察糖尿病肾病（DN）大鼠经雷公藤多苷干预后,肾皮质中结缔组织生长因子（connective tissue growth factor,CTGF）表达的变化,并探讨雷公藤多苷在其中的作用。方法：Wister大鼠48只,随机分为正常对照组、DN组与雷公藤多苷治疗组。DN组与雷公藤多苷治疗组大鼠分别给予链脲佐菌素（STZ）55mg／kg一次性腹腔注射建立糖尿病大鼠模型。模型建立后第4、8、12周各组分别处死4只大鼠并收集标本,记录体重、检测血糖、血肌酐、尿素氮、胆固醇、三酰甘油、24h尿蛋白定量。用RT-PCR法检测肾皮质CTGF mRNA含量表达水平。结果：（1）DN组大鼠血糖、血肌酐、血尿素氮、血胆固醇、血三酰甘油、24h尿蛋白定量均明显高于正常对照组（P〈0．01）;肾组织CTGFmRNA的表达量明显高于正常对照组（P〈0．01）。（2）雷公藤多苷治疗组血糖、血肌酐、血尿素氮、血胆固醇、血三酰甘油及24h尿蛋白定量在第12周与DN组比较均有统计学差异（P〈0．01）;治疗组肾组织CTGFmRNA含量的表达明显低于DN组（P〈0．01）。结论：雷公藤多苷可减少DN大鼠尿蛋白的排泄及降低血尿素氮、血肌酐,明显降低肾组织CTGFmRNA表达量,提示雷公藤多苷能有效延缓DN进展,并有一定的保护作用。     

雷公藤多苷对早期糖尿病肾病大鼠肾小管上皮细胞Smad2的影响

目的：探讨雷公藤多苷（TW）对糖尿病肾病（DN）模型大鼠肾组织中Smad2表达的影响。方法：用TW灌服DN模型大鼠,生化检测实验大鼠24 h尿蛋白、血肌酐（Scr）、尿素氮（BUN）、血浆白蛋白（Alb）;Leica Qwin Plus图像分析系统测量肾小球直径;PAS染色观察大鼠肾脏病理;免疫组化（IHC）检测肾组织Smad2表达。结果：TW能够降低实验大鼠肾小管上皮细胞Smad2的表达及24 h尿蛋白、Scr、BUN,升高Alb,减少肾小球直径及肾重,改善肾脏病理,与模型组相比,P〈0.05或P〈0.01。结论：TW可能通过影响DN大鼠肾小管上皮细胞Smad2的表达,而具抗肾间质纤维化的作用。     

小剂量雷公藤多苷对糖尿病肾病蛋白尿影响的临床观察

虽然目前对糖尿病肾病还缺乏行之有效的治疗措施,但降低尿蛋白无疑是有效延缓糖尿病肾病发展、肾功能恶化的重要手段之一。本研究在糖尿病肾病常规治疗基础上加用小剂量雷公藤多苷（0.5 mg·kg-1·d-1）以观察其对糖尿病肾病蛋白尿的作用及安全性。     

雷公藤多苷片对糖尿病肾病患者大量蛋白尿的影响

目的：观察雷公藤多苷片对糖尿病肾病患者临床症状、蛋白尿、体液免疫的影响。方法：60例2型糖尿病患者，符合Mogensen糖尿病肾病诊断标准，选择尿蛋白排出率〉1500mg／24h，血清肌酐（Scr）〈442μmol／L早中期病例为研究对象。随机分为2组，雷公藤多苷片治疗组及科素亚对照组。疗程2个月，观察患者临床症状的变化，检测24h尿蛋白定量，IgA、IgM、IgG、C3、C4，肝肾功能，血白细胞。结果：治疗组明显改善乏力、腰酸、水肿等症状，减少尿蛋白，可提升血清白蛋白、IgG、IgA、C3、C4水平。结论：雷公藤多苷片可明显减少糖尿病肾病蛋白尿，缓解症状，提高免疫球蛋白和补体水平，改善患者免疫功能。     

雷公藤多苷治疗糖尿病肾病的利与弊

糖尿病肾病（ diabetic nephropathy,DN）为糖尿病最常见的慢性并发症之一,也是导致终末期肾衰竭（ end stage of renal disease,ESRD）最主要的原发疾病。血管紧张素转换酶及其受体抑制剂（ ACEI／ARB）在糖尿病肾病中的应用已成为共识,但由于临床疗效有限,不少学者仍在积极寻求更有效的方法。其中,富有中医药特色的雷公藤多苷以其独特的作用受到关注。下面笔者结合文献和个人经验,谈谈雷公藤多苷在DN的治疗应用。     

益肾调经方对阿霉素肾病伴雷公藤多苷致性腺抑制的雌性大鼠肾保护作用的实验研究

目的：在阿霉素尾静脉注射造成微小病变肾病动物模型的基础上，建立雷公藤多苷致雌性肾病大鼠性腺抑制的模型，并观察益肾调经方（YSTJF）对其的影响，探讨YSTJF对阿霉素肾病伴性腺抑制的雌性大鼠肾保护作用。方法：雌性SD大鼠50只，随机挑选6只作正常对照组。余用阿霉素5mg／kg一次性尾静脉注射造成微小病变肾病动物模型，并随机分为6组：（1）肾病模型组：再予蒸馏水灌胃。（2）性腺抑制组：再予雷公藤多苷80mg·kg^-1·d^-1。（3）YSTJF高、中、低剂量组：予雷公藤多苷80mg·kg^-1·d^-1、YSTJF水煎浓缩液（分别为含生药75、50、25g·kg^-1·d^-1）合用每日灌胃。（4）西药对照组：予雷公藤多苷片80mg·kg^-1·d^-1和结合雌激素倍美力0．02mg·kg^-1·d^-1叫合用每日灌胃。该实验共8周，实验结束后收集标本，24h尿蛋白、N-乙酰-β-D-氨基葡萄糖苷酶（NAG），血肌酐（Scr），尿素氮（BUN）；光镜下观察肾脏组织的病理。结果：肾病模型组的尿蛋白、尿NAG酶明显高于正常对照组；性腺抑制组、YSTJF各剂量组和西药对照组能降低尿蛋白、尿NAG酶；、YSTJF中低剂量组降低尿蛋白与性腺抑制组比较（P〈0．05）；、YSTJF高剂量组降低尿NAG酶优于低剂量组和性腺抑制组。肾病模型组BUN高于正常对照组（P〈0．05）；性腺抑制组、、YSTJF各剂量组和西药对照组均低于肾病模型组（P〈0．05）；YSTJF各剂量组、西药对照组比性腺抑制组降低更明显（P〈0．05），YSTJF各剂量组比西药对照组为佳（P〈0．05）。从肾脏病理改变的小管间质病变积分分析：YSTJF各剂量组积分低于肾病模型组、性腺抑制组（P〈0．05），高、中剂量组较西药对照组为佳。结论：YSTJF可以通过降低尿蛋白、NAG酶、BUN，降低肾小管间质积分，改善肾小管间质病理改变，对于阿霉素肾病伴性腺抑制的雌性大     

雷公藤多苷对糖尿病肾病患者转化生长因子β1的影响

目的:观察雷公藤多苷(TL)对糖尿病肾病(DN)患者血清和尿液转化生长因子-β1(TGF-β1)的影响,探讨TL对DN的治疗的作用.方法:87例DN患者分为两组,治疗组(45例)采用TL和糖尿病(DM)常规治疗;对照组(42例)单用DM常规治疗,疗程3个月,观察患者治疗前后血清和尿液TGF-β1等指标的变化.结果:DN患者血清和尿液TGF-β1水平显著高于健康人(P<0.01),且与尿蛋白呈显著正相关(P<0.01).治疗组治疗后血清和尿液TGF-β1、尿蛋白水平显著降低(P<0.01,P<0.01,P<0.05)且血清和尿液TGF-β1降低与尿蛋白降低呈显著正相关(P<0.01).结论:TL能明显降低DN患者血清尿液TGF-β1和尿蛋白水平,对DN患者肾功能有一定的保护作用.     

雷公藤多甙对白细胞介素-1β诱导大鼠滑膜细胞株RSC-364基质金属蛋白酶-3、c-Jun基因表达的影响

目的 观察雷公藤多甙对白细胞介素(IL)-1β诱导大鼠滑膜细胞株RSC-364基质金属蛋白酶-3(MMP3)、c-Jun mRNA表达的影响.方法 选择不同浓度的雷公藤多甙作用IL-1β刺激的大鼠滑膜细胞株RSC-364,37℃,5%CO2的环境下培养48 h,运用酶联免疫吸附试验(ELISA)检测细胞上清的MMP3和实时聚合酶链反应(real-time PCR)检测分析细胞中的c-Jun mRNA表达.结果 不同浓度的雷公藤多甙(0、5、10、20 mg/L)对IL-1β刺激的大鼠滑膜细胞株RSC-364分泌MMP3的水平依次是198、176、140、115 μg/L,c-Jun mRNA表达分别是1.17×106、3.31×105、9.32 ×104、2.81 ×104copies/ml,即呈现明显地抑制.重要的是:c-Jun mRNA表达抑制呈现明显的剂量依赖性.结论 雷公藤多甙治疗类风湿性关节炎的有效的作用机制,可能与其抑制MMP3、c-Jun基因的表达相关.     

Effect of recurrent pregnancies on the evolution of adriamycin nephropathy

BACKGROUND.: In rats with incipient adriamycin nephropathy, pregnancy increasesurine protein excretion and mean arterial pressure, with nochanges in the glomerular filtration rate. Renal histology isnormal and the glomerular TxB₂/PGE2 ratio is increased. METHODS.: In the present study we evaluated the influence of repeatedpregnancies on the evolution of adriamycin nephropathy. Twoweeks after a first delivery, rats were mated again and werefollowed till 35 days after the second delivery. RESULTS.: In pregnant rats with adriamycin nephropathy, urine proteinexcretion and mean arterial pressure returned to values identicalto those found in age-sex-matched virgin rats with adriamycinnephropathy. At the end of the second pregnancy, mean arterialpressure and urine protein excretion were again elevated, comparedwith virgin rats with adriamycin nephropathy. Thirty-five daysafter the second delivery, urine protein excretion and meanarterial pressure remained elevated, 296±50 mg/day vs115±26 and 121±4 vs 110±1 mmHg respectively,P<0.05. Glomerular nitration rate remained unchanged 0.84±0.09vs 0.79±0.09 ml/min in virgin rats with adriamycin nephropathy.The glomerular TxB₂/PGE₂ ratio was decreased, contrasting withthe first pregnancy. At the end of the second pregnancy, histologicalexamination of the kidneys in rats with adriamycin nephropathyrevealed a significant increase in mesangial expansion. It waseven more marked 35 days later, at the last follow-up, witha semiquantitative score of 162±29 vs 81±20 invirgin adriamycin nephropathy rats, P<0.05. CONCLUSIONS.: In rats with adriamycin nephropathy, repetitive pregnanciesseem to aggravate the natural course of the disease in an irreversiblefashion. The earlier changes in glomerular prostanoid synthesis,particularly on thromboxane, may play a pathogenic role by activatingmesangial cell matrix synthesis.     

Effect of allopurinol in the course of adriamycin induced nephropathy

The role of superoxide in adriamycin-induced nephropathy (single dose; i.v. 3 mg/kg) has been studied by blocking superoxide synthesis through the administration of allopurinol (500 mg/L in drinking water). In Experiment I (EI), allopurinol administration was started 3 days prior to nephropathy induction and continued until day 14. In Experiment II (EII) allopurinol administration was started 2 weeks after nephropathy induction and was maintained until the end of the experiment (26 weeks). Affected glomeruli frequency and tubulointerstitial lesion index (TILI) were determined at Weeks 2 and 4 (EI) and Week 26 (EII). In EI, the 24 h mean proteinuria in the nephrotic control group (NCG-I) differed from that of the treated nephrotic group (TNG-I) at Week 1 (TNG = 33.3 +/- 6.39 mg/24 h; NCG = 59.8 +/- 6.3 mg/24 h; p < 0.05) and 2 (NCG-I = 80.0 +/- 17.5 mg/24 h; TNG-I = 49.1 +/- 8.4 mg/24 h; p < 0.05). No glomerular alterations were observed and TILI medians were not different in both nephrotic groups at week 2 (NCG-I = 1+: TNG = 1+) and 4 (NCG = 4+; TNG = 4+). In EII, NCG-II and TNG-II presented different 24 h proteinuria values only at Week 6, (136.91 +/- 22.23 mg/24 h and 72.66 +/- 10.72 mg/24 h, respectively; p < 0.05). Between nephrotic groups, there was no statistical difference in the median of affected glomeruli (CNG-II = 56%; TNG-II = 48%) and TILI (NCG-II = 8+; TNG-II = 9+). Thus, allopurinol was associated with a transient reduction in proteinuria and it did not alter the progression of the nephropathy.     

Effect of mycophenolate mofetil on the progression of adriamycin nephropathy.

In order to assess the effects of mycophenolate mofetil (MMF) on the development of adriamycin-induced nephropathy, the development of this nephropathy in rats treated with MMF was compared to that in non-treated animals (group ADR + V) over 28 weeks. At weeks 8, 16 and 20, 24-h proteinuria of the treated group statistically differed from that of the non-treated group. However, no significant difference in proteinuria was observed thereafter between the groups. At the end of the experiment, there was no significant difference between both groups regarding the frequency of glomerular lesion (Group ADR + V: Md = 35%, P25 = 20%, P75 = 68%; Group ADR + MMF: Md = 27%, P25 = 9.5%, P75 = 54%); tubulointerstitial lesion index (Group ADR + V: Md = 7, P25 = 1.5, P75 = 9; Group ADR + MMF: Md = 8, P25 = 2, P75 = 9); glomerulosclerosis area (group ADR + V = 2779 microm2, P25 = 751.8 microm2, P75 = 3115 microm2; Group ADR + MMF = 1147 microm2, P25 = 3969.7 microm2, P75 = 1560 microm2); and, interstitial fibrosis area (Group ADR + V: Md = 218200 microm2, P25 = 78670 microm2, P75 = 282700 microm2 group ADR + MMF: Md = 136000, P25 = 25010, P75 = 255800 microm2). In conclusion, MMF caused a temporary reduction in proteinuria but did not change the severity of the renal lesion observed after 28 weeks.     

Effects of cyclosporine on adriamycin induced nephropathy

SUMMARY: The effect of cyclosporine on adriamycin nephropathy was studied over both short (6 weeks) and long (26 weeks) periods. In the short-term study, cyclosporine was introduced 2 weeks after nephritis induction and in the long-term study, 14 weeks after the first adriamycin injection. the animals with adriamycin nephropathy treated with cyclosporine, studied for 6 weeks, developed proteinuria with increasd renal size and glomerular area. the nephrotic animals treated with cyclosporine showed less proteinuria than the nephrotic control animals. There were no differences in glomerular area, creatinine clearance or serum creatinine and kidney weight between the treated nephrotic group and the health control group. the nephrotic animals, studied for the longer period, developed intense proteinuria, decreased creatinine clearance, glomerular necrosis and sclerosis, severe tubulointerstitial nephritis, increased hydroxyproline concentration and tubulointerstitial area. No difference was observed between the nephrotic animals treated with cyclosporine and those not treated. In conclusion, Cyclosporine A reduced proteinuria, glomerular hypertrophy and kidney weight in rats with short-term nephropathy but had no effect on the established nephropathy.     

Suppressive effect of superoxide dismutase on adriamycin nephropathy.

A single intravenous injection of adriamycin (ADR) results in marked proteinuria and glomerular morphological changes that are similar to minimal-change disease in humans. We examined the effect of superoxide dismutase (SOD) on ADR-induced proteinuria. ADR in a dose of 7.5 mg/kg body weight significantly increased urinary protein by day 14; proteinuria rapidly increased thereafter. Concurrent administration of SOD (50 mg/kg) over 30 min prior to and 30 min following ADR injection markedly reduced proteinuria. Twenty-one days after the treatment with SOD, the amount of urinary protein was 108.6 +/- 43.1 mg/24 h in the experimental animals, while it was 221.6 +/- 102.9 mg/24 h in the ADR control group (p less than 0.05). There were also less severe glomerular morphologic changes in the SOD group versus ADR controls. The protective effects of SOD provide indirect evidence that oxygen free radicals are important mediators of ADR-induced proteinuria.     

贝那普利对大鼠环孢素A慢性肾毒性模型转化生长因子-β1的影响

目的 探讨贝那普利防治环孢素A(CsA)慢性肾毒性的机理。方法 给进低盐饮食的SD大鼠灌服CsA，28d后用逆转录聚合酶链反应(RT-PCR)的方法观察肾内转化生长因子-β1(TGF-β1)及肾素mRNA表达，用免疫组化测定肾组织中TGF-β1及四型胶元(Collagen Ⅳ)的改变，并观察贝那普利对上述指标的影响。结果 使用CsA的大鼠肾内TGF-β1及肾素mRNA呈现高表达，并伴有Collagen Ⅳ肾内沉积增加；加用贝那普利能抑制TGF-β1的表达，但对肾素mRNA表达的影响不明显。结论 贝那普利对CsA的慢性肾毒性的防治与其抑制TGF-β1 mRNA的高表达及减轻肾内CollagenⅣ的沉积有关。     

他克莫司对阿霉素肾病大鼠足细胞损伤的修复作用

目的探讨他克莫司对阿霉素肾病大鼠足细胞损伤的修复作用。方法通过阿霉素尾静脉注射建立大鼠微小病变肾病模型,并以他克莫司进行干预。大鼠随机分为对照组、模型组和他克莫司组。每组分别于第0、7、14、21、28、35天采集尿液,检测尿蛋白排泄量,于造模第35天处死大鼠收集肾脏标本。电镜观察各组大鼠足细胞足突融合情况;TUNEL检测法对肾小球进行染色;免疫组化或免疫荧光对WT-1、caspase-3进行定位和半定量检测,观察大鼠足细胞的数量和凋亡。结果阿霉素尾静脉注射后大鼠尿蛋白显著增加,足细胞数目明显减少,足突广泛融合;与模型组比较,他克莫司组大鼠24 h尿蛋白明显减少,足突融合改善,足细胞数目增多。与对照组比较,模型组大鼠肾小球内caspase-3蛋白表达增加,TUNEL染色加深;与模型组比较,他克莫司组肾小球内caspase-3表达量下降,TUNEL阳性率减少,说明足细胞凋亡减少。结论他克莫司可修复阿霉素肾病大鼠足细胞损伤,其机制与抑制足细胞凋亡有关。