肾康注射液对糖尿病肾病大鼠细胞外基质影响的实验研究

目的:探讨肾康注射液(SKI)对DN肾小球硬化的防治作用及机制.方法:采用单肾切除加链脲佐菌素诱导的方法制造大鼠糖尿病肾病(DN)模型,设置高低两种剂量的肾康注射液治疗组,洛汀新对照组,模型组和正常组.观察各组肾小球中的细胞外基质(ECM)不同组份的变化.结果:DN大鼠肾小球中ECM成份FN、LN和CoⅣ的合成和分泌明显增多.SKI能明显抑制肾小球合成和分泌FN、LN和CoⅣ,其药物作用优于洛汀新,并且与剂量呈依赖关系.结论:减少肾小球合成和分泌FN、LN和CoⅣ是SKI防治DN肾小球硬化的作用机制之一.     

肾康注射液对缺血再灌注急性肾损伤小鼠的作用探讨

目的探讨肾康注射液对缺血再灌注致急性肾损伤小鼠的保护作用。方法随机选取相同数量C57BL/6小鼠,夹闭双侧肾动脉不同时间(15、25、45 min),以制备轻、中、重度缺血再灌注急性肾损伤小鼠模型,并根据造模后立即腹腔注射10 m L/kg肾康注射液(SKI)或10 m L/kg无菌生理盐水(NS)下分为6组各6只:Ⅰ轻度损伤+SKI干预组;Ⅱ轻度损伤+NS组;Ⅲ中度损伤+SKI组;Ⅳ中度损伤+NS组;Ⅴ重度损伤+SKI组;Ⅵ重度损伤+NS组;同时设置一组空白对照组(Ⅶ组):只开关腹,不做夹闭肾动脉处理。干预24 h后,处死小鼠取标本,予生化法检测血肌酐(Scr)、血尿素氮(BUN)、组织超氧歧化酶(SOD)及丙二醛(MDA);HE染色法观察肾小管坏死、免疫组织化学染色法观察肾小管细胞凋亡及中性粒细胞浸润;实时PCR检测组织炎症因子表达。结果 SKI干预后轻、中度肾损伤小鼠Scr下降,轻度损伤+SKI组对比轻度损伤+NS组,下降明显(P0.05);中度损伤+SKI组比中度损伤+NS组Scr明显下降(P0.05)。同时轻、中度模型组肾组织病理损伤也有所改善,但重度损伤组均无改善。进一步机制探究发现SKI干预后氧化应激改善:SKI干预组较NS干预组肾组织SOD活性更高,MDA则更低(P0.01)。在组织炎症方面,SKI干预后组织浸润的中性粒细胞减少,i NOS、IL-6表达下降。此外肾小管凋亡减轻。结论SKI可以改善缺血再灌注急性肾损伤小鼠肾功能,并逆转一定程度的病理损伤,其机制与改善氧化应激、炎症反应相关。     

肾康注射液对腹膜透析液诱导的大鼠腹膜组织形态及TNF-α,TGF-β1的影响

目的:研究肾康注射液(Shenkang injection,SKI)对腹膜透析液(peritoneal dialysis solution,PDS)诱导的大鼠腹膜组织形态及肿瘤坏死因子-α(TNF-α),转化生长因子-β1(TGF-β1)影响。方法:50只SD大鼠,随机分为正常组,4.25%PDS组和SKI低、中、高剂量组;除正常组不注射PDS外,其余4组分别ip 4.25%PDS,低、中、高剂量的SKI+4.25%PDS。于注射8周后,水合氯醛麻醉各组大鼠,同时沿腹白线剪开腹壁,量取腹腔内的透出液,ELISA法测定透出液中TNF-α的含量。取壁层腹膜组织,观察腹膜组织形态改变,并用免疫组化法检测TGF-β1的表达。结果:与正常组比较,PDS组大鼠透出液中TNF-α含量和壁层腹膜间皮细胞的TGF-β1表达显著升高(P0.01),且腹膜间皮细胞重度肿胀变性、脱落,间皮下基质明显增生,并可见大量血管增生及炎细胞浸润。与PDS组比较,SKI中、高剂量组的TNF-α含量显著降低(P0.01),SKI各剂量组壁层腹膜间皮细胞TGF-β1的表达明显减少(P0.05),且腹膜结构改变减轻、炎症细胞浸润和基质增生减少。结论:腹膜透析液中加入SKI具有改善PDS诱导腹膜纤维化,其作用机制可能与抑制TNF-α,TGF-β1的增加有关。     

中医药防治糖尿病肾病的机理研究现状

糖尿病肾病(diabeticnephropathy,DN)是糖尿病最严重和最常见的慢性并发症之一,1型(IDDM)及2型(NIDDM)糖尿病患者肾脏受累率分别为30%和20%[1]。DN的病理特征是:肾小球基底膜(GBM)增厚、膜基质的扩张和细胞外基质(ECM)的重构。DN临床病程可简单概括为:尿微量白蛋白前期、尿微量白蛋白期、尿蛋白期、肾功能不全期、终末肾衰期。随着DN的发病率逐年上升,DN愈来愈成为导致终末期肾衰(ESRD)的主要原因。中医药在防治DN的研究方面做了大量的工作,本文仅就中医药防治DN的实验研究机理综述如下。1纠正与高糖相关的生化代谢紊乱1.1抑…     

大黄治疗糖尿病肾病的研究近况

通过对相关临床文献的搜集、整理及归纳,本文从糖尿病肾病(DN)的发病机制、大黄的相关药理作用及大黄治疗糖尿病肾病(DN)的研究与应用3个方面进行分析、研究。结果表明:大黄能有效减少肠道中氨基氮的重吸收,改善氮质血症,抑制残余肾组织的代偿性肥大,抑制肾小球系膜细胞的增殖、纠正脂代谢紊乱,减少蛋白尿,改善微循环、抗凝、抗血栓等降低微血管并发症的相关作用。由此可见,中药大黄及其提取物对DN有较好的疗效,并取得了可喜的成绩。现笔者就相关文献作一综述。     

雷公藤甲素对糖尿病肾病小鼠肾足细胞自噬及调亡的影响

目的:探讨雷公藤甲素对糖尿病肾病(DN)小鼠肾足细胞自噬及调亡的影响。方法:选择小鼠60只,随机分为DN组和雷公藤甲素组各30只,光镜下观察各组肾组织病理学改变情况,观察肾功能变化情况,对其行行威廉姆斯肿瘤蛋白1(WT-1)免疫组化染色,观察足细胞情况,透射电镜观察足细胞自噬情况,采用Western blot法检测肾组织中Podocin、Lc3、Bax及Caspase-3的表达情况。结果:PAS染色结果显示,DN组小鼠系膜基质明显增多,雷公藤甲素组小鼠系膜基质明显减少。WT-1免疫组织化学染色显示,雷公藤甲素组肾足细胞数量比DN组明显增多。Western Blot结果显示,与DN组小鼠比较,雷公藤甲素组Podocin表达明显增加,LC3-Ⅱ/LC3-Ⅰ比值明显上升,凋亡相关蛋白Bax及Caspase-3表达明显减少(P 0.05)。结论:雷公藤甲素可减轻足细胞损伤延缓DN的进展,可DN的治疗起到参考价值。     

当归补血汤对糖尿病大鼠肾组织内质网IRE1α-JNK通路的抑制作用

目的:为了研究当归补血汤对糖尿病肾病(diabetic nephropathy,DN)大鼠肾组织内质网应激通路IRE1α-JNK的作用机制及其对高糖下肾脏组织的保护作用。方法:链脲佐菌素(STZ)联合高脂饲料,建立糖尿病肾病动物模型,分为:正常对照组(N)、糖尿病肾病对照组(DN)、格列奇特组(GT)、当归补血汤低剂量组(DL)、当归补血汤高剂量组(DH)。对照组(N、DN)以生理盐水灌胃,治疗组分别以格列齐特缓释片溶液、高低浓度当归补血汤浓缩液灌胃处理8周后取材,用酶偶联比色法检测相关生化指标(血糖、血脂及肾功能指标),光镜下观察肾脏病理改变;West Blog、RT-PCR检测IRE1α、JNK蛋白的表达,TUNEL法测定各组肾脏组织细胞的凋亡。结果:结果显示,DN组大鼠空腹血糖、血生化指标较N组明显升高;肾组织中p-IRE1α,p-JNK蛋白表达量较对照组N明显增加,细胞凋亡程度高;治疗组血糖、血脂、血肌酐等指标均低于DN组;肾脏p-IRE1α,pJNK蛋白表达较DN组下降;肾脏组织中细胞凋亡较DN组改善,N组与DH组无明显差异(P0.05)。结论:研究表明当归补血汤可以抑制高糖下肾组织IRE1α-JNK通路的表达,减轻高糖下肾脏的内质网应激反应,从而保护肾组织。     

绞股蓝总皂苷对糖尿病肾病大鼠足细胞损伤的影响及机制

目的:研究绞股蓝总皂苷对糖尿病肾病(DN)大鼠肾脏足细胞相关分子(nephrin)、血管通透因子(VEGF)mRNA表达的影响,探讨其降低尿蛋白、保护肾脏的机制。方法:采用单侧肾切除加链脲佐菌素(STZ)注射法改良复制DN模型,实验分为正常组、模型组、治疗组(绞股蓝总皂苷高、中、低剂量组)和缬沙坦组。干预4周后电镜观察肾小球超微结构改变,RT-PCR检测nephrin、VEGF mRNA表达。结果:各治疗组病理变化较模型组均有不同程度改善:足突增宽或部分融合,基底膜增厚减轻,同时nephrin mRNA表达水平较模型组明显上调(P0.01),VEGF表达明显抑制(P0.01),其中高剂量组和缬沙坦组效果类似,明显优于低剂量组(P0.01)。结论:绞股蓝总皂苷可能通过上调足细胞相关分子nephrin表达,抑制分泌VEGF过表达,减轻足细胞超微结构改变,从而保护足细胞,降低尿蛋白,延缓肾小球硬化。     

肾康注射液调控ERK1/2/MMPs信号通路促进肾衰竭模型鼠细胞外基质降解的作用和机制

探讨肾康注射液(Shenkang injection,SKI)在体内调控细胞外信号调节激酶(extracellular-signal regulated protein kinase,ERK)1/2/基质金属蛋白酶(matrix metalloproteinases,MMPs)信号通路而改善肾衰竭模型鼠细胞外基质(extracellular matrix,ECM)降解的作用和机制。将20只大鼠随机分为假手术组、模型组、SKI组、马来酸依那普利(enalapril maleate,EM)组。采用腺嘌呤灌胃联合单侧输尿管结扎术(unilateral ureteral obstruction,UUO)建立肾衰竭模型。造模后,SKI组和EM组大鼠分别经腹腔注射或灌胃给予SKI或EM悬浊液,其余2组大鼠经灌胃给予蒸馏水,共3周;其间,检测各组大鼠24 h尿蛋白排泄量(urinary protein excretion,Upro)和尿N-乙酰-β-D-氨基葡萄糖苷酶(urinary N-acety1-β-D-glucosaminidase,UNAG);给药3周后,处死全部大鼠,抽取血液,摘除双肾,观察肾组织形态特征,检测血清生化指标和肾组织IV型胶原(collagen type IV,CIV),MMP-2,MMP-9,金属蛋白酶组织抑制剂(tissue inhibitors of metalloproteinase,TIMP)-1,ERK1/2以及磷酸化ERK1/2(phosphorylated-ERK1/2,p-ERK1/2)蛋白表达量。结果表明,经SKI干预后,模型鼠血清肌酐(serum creatinine,Scr),血清尿素氮(blood urea nitrogen,BUN),尿酸(uric acid,UA),白蛋白(albumin,Alb),Upro,UNAG以及肾脏组织形态均得到不同程度的改善,这些作用与EM相仿;SKI还可以调节模型鼠肾组织MMP-2,MMP-9,TIMP-1蛋白表达,下调p-ERK1/2蛋白表达,这些作用不同于EM。总之,SKI在体内可能是通过调控肾组织ERK1/2信号通路活性,干预MMPs/TIMP-1表达,促进ECM降解,延缓肾衰竭进展。     

2型糖尿病肾病患者血清胱抑素C与动脉粥样硬化的关系

目的:探究2型糖尿病肾病(DN)患者血清胱抑素C(CysC)、超敏C反应蛋白(hs–CRP)与动脉粥样硬化的关系。方法:选取2018年7月至2019年6月东莞市第三人民医院收治的2型糖尿病(DM)患者80例及健康志愿者30例作为研究对象,检测50例2型DN(非透析)患者、30例单纯2型DM患者(无并发症)及30例健康志愿者血清CysC、hs–CRP的含量。彩色多普勒超声测量颈动脉内中膜厚度(IMT)。结果:2型DN组的CysC、hs–CRP水平明显高于DM组及健康对照组,差异具有统计学意义(P 0.05);相关分析结果发现CysC、hs–CRP均与2型DN患者颈动脉IMT呈正相关。结论:2型DN患者血清CysC、hs–CRP与颈动脉IMT呈正相关,可能促进2型DN患者动脉粥样硬化的发生发展。     

Shenkang Injection protects against diabetic nephropathy in streptozotocin (STZ)-induced mice through enhancement of anti-oxidant and anti-inflammatory activities

ObjectiveTo investigate the protective effects and possible mechanisms of Shenkang Injection (SKI) on the diabetic nephropathy in streptozotocin-induced mice.MethodsSTZ with the feeding of high fat diet (HFD) was used to induce diabetic mice. The balb/c mice and diabetic mice were then randomly divided into five groups: (1) control group, (2) model group, (3) alprostadil (Alp, 1.5 μg/kg) group, (4) SKI (30 ml/kg) group, (5) Alp (1.5 μg/kg) + SKI (15 ml/kg) group. After six weeks' treatment, blood, urine and kidney tissues were collected for biochemical assay, ELISA assay, and pathological analysis.ResultsDiabetic mice exhibited evident manifestations of diabetic nephropathy (DN), as indicated by increased 24-h urine volume, urinary albumin and kidney weight index (P < 0.01), which could be attenuated by SKI treatment (P < 0.01). SKI was further found to improve abnormal morphology in glomerulus with increased glomerular volume and to decrease urinary N-acetyl-b-D-glucpsaminidase (NAG), β2-microglobulin (β2-MG), and kidney injury molecules-1 (KIM-1) levels (P < 0.05, P < 0.01). Plasma levels of anti-oxidant enzymes significantly reduced in the diabetic mice, and those decreases could be reversed by SKI and Alp treatments. Additionally, SKI obviously suppressed the diabetes-induced increases of pro-inflammatory cytokines (IL-6, IL-1β and TNF-α) (P < 0.01). Meanwhile, SKI was found to effectively attenuate the diabetes-induced coagulation dysfunction, as evidenced by lengthening prothrombin and thrombin time, and decreasing plasma levels of fibrinogen (FIB), 6-K-PGF1α and thromboxane B2 (TXB2) (P < 0.05, P < 0.01). With SKI and Alp combined treatment, the anti-oxidant activities and improvements of coagulation dysfunction were enhanced.ConclusionSKI possesses a remarkable property to prevent diabetic nephropathy. The improvements of kidney function and hypercoagulability by SKI were enhanced with Alp combined treatment. The molecular mechanisms underlying the protection of SKI against DN may be related to enhancing the anti-oxidant and anti-inflammatory activities, and improving the coagulation dysfunction.     

肾康注射液对肾小球系膜细胞抑制作用的血清学研究

目的探讨肾康注射液与苯那普利对人肾小球系膜细胞(MC)增殖的不同抑制作用强度。方法采用体外培养的人肾小球系膜细胞技术和血清药理学方法，观察具有降逆泄浊、益气活血作用的中药肾康注射液及苯那普利对MC增殖的影响。结果肾康注射液与苯那普利对MC增殖均具有抑制作用，但在等效剂量浓度下，肾康注射液对MC的抑制作用优于苯那普利。结论MC是肾康注射液发挥治疗作用的重要靶细胞，抑制MC增殖可能是该方延缓肾小球硬化的重要机制之一。     

肾康注射液对肾小球系膜细胞抑制作用的血清学研究

目的：探讨肾康注射液与苯那普利对人肾小球系膜细胞（MC）增殖的不同抑制作用强度。方法：采用体外培养的人肾小球系膜细胞技术和血清药理学方法,观察具有降逆泄浊,益气活血作用的中药肾康注射液及苯那普利对MC增殖的影响。结果：肾康注射液与苯那普利对MC增殖均具有抑制作用,但在等效剂量浓度下,肾康注射液对MC的抑制作用优于苯那普利,结论：MC是肾康注射液发挥治疗作用的重要靶细胞,抑制MC增殖可能是该方延缓肾小球硬化的重要机制之一。     

肾康注射液与苯那普利对肾小球系膜基质作用的对比研究

目的 探讨肾康注射液与苯为利对人肾小球系膜细胞外基质（ＥＣＭ）不同成分的抑制作用强度。方法 采用体外培养的人肾小球系膜细胞技术，观察具有降逆泄浊、益气活血功效的中药肾康注射液及苯那普利时ＥＣＭ不同成分的影响。结果 肾康注射液具有抑制ＥＣＭ成分中纤维连接蛋白（ＦＮ）、层粘连蛋白（ＬＮ）和Ⅳ型胶原（ＣｏｌⅣ）的作用，该抑制作用具有一定的量效关系。苯那普利对ＥＣＭ中ＦＮ、ＣｏｌⅣ也有一定的抑制作用，但不     

Shenkang injection improves coagulation in patients with chronic kidney disease: a systematic review and Meta-analysis

OBJECTIVE: To investigate the effect of Shenkang injection(SKI) on chronic kidney disease(CKD).METHODS: Seven databases including Cochrane Central Register of Controlled Trials, PubMed, EMBASE, MEDLINE, China National Knowledge Infrastructure, Wanfang Database, and CQVIP from their inception to March 2018 were searched. Only randomized controlled trials that evaluated conventional treatment and conventional treatment with SKI in CKD patients were investigated. Outcomes such as fibrinogen(FIB), D-dimer, prothrombin time(PT), activated partial thromboplastin time(APTT), and the side effects of SKI were analyzed using Revman 5.3 software. The quality of the studies was assessed using the Cochrane Collaboration's Risk of Bias tool and the quality of evidence was assessed using GRADEpro.RESULTS: Four randomized controlled trials were investigated in our analysis, and these studies were of moderate quality. For FIB and D-dimer, SKI had a superior effect compared with the control group[mean difference(MD)=-1.23, 95% confidence interval(CI):-1.46,-0.99, P 0.01; MD =-0.36, 95%CI:-0.51,-0.21, P 0.01, respectively]. SKI increased APTT and PT compared with the control(MD = 7.34, 95% CI: 3.05, 11.62, P 0.01; MD = 3.40,95% CI: 2.2, 4.61, P 0.01, respectively). In the four studies, there were no side effects that were related to SKI.CONCLUSION: SKI may be effective in improving coagulation in patients with CKD without obvious adverse reactions. However, more well-designed studies are required to confirm the findings.     

Effect of SKI 306X, a new herbal anti-arthritic agent, in patients with osteoarthritis of the knee: a double-blind placebo controlled study.

SKI 306X is a purified extract from a mixture of three oriental herbal medicines (Clematis mandshurica, Trichosanthes kirilowii and Prunella vulgaris) that have been widely used for the treatment of inflammatory diseases such as lymphadenitis and arthritis in far East Asia. A double-blind, controlled study was performed to evaluate the efficacy and safety of SKI 306X with placebo in 96 patients with classical osteoarthritis of the knee. Patients were randomized to four treatment groups: placebo, 200 mg, 400 mg and 600 mg of SKI 306X t.i.d.. Clinical efficacy and safety were evaluated for 4 weeks continuous treatment. SKI 306X demonstrated its clinical efficacy, as assessed by 100 mm visual analogue scale (VAS), Lequesne index and patients' and investigators opinion of the therapeutic effect compared with placebo (p<0.01). No significant adverse events were observed in patients treated with SKI 306X. This study demonstrated that SKI 306X, a new herbal anti-arthritic agent provided clinical efficacy in patients with osteoarthritis.     

肾康注射液对肾小管上皮细胞转化生长因子-βmRNA基因表达的影响

目的：探讨肾康注射液对体外培养的肾小管上的细胞LLC-PK1转化生长因子-βmRNA（TGF-βmRNA）基因表达的影响。方法：用逆转录聚合酶链反应（TR-PCR）、Northern印迹杂交（Northern blot）方法，以单味大黄注射液为实验对照组，检测肾康注射液对肾小管上皮细胞TGF-βmRNA基因表达的影响。结果：肾康注射液可以明显下调肾小管上皮细胞TGF-βmRNA基因的表达，并呈剂量依赖关系，肾康注射液作为明显优于同等含量的单味大黄注射液。结论：肾康注射液下调肾小管上皮细胞TGF-βmRNA基因表达，是其防治慢性肾功能衰竭（CRF）的机理之一。     

Clematis mandshurica protected to apoptosis of rat chondrocytes

OBJECTIVE: To investigate the effect of SKI 306X, a purified extract from the mixture of three herbs, i.e. Clematis mandshurica, Trichosanthes kirilowii and Prunella vulgaris, on apoptosis in chondrocytes. DESIGN: Rat chondrocyte cell line RCJ3.1C.18 cells were incubated with 1 microM staurosporin and SKI 306X or each of its components. Cell viability was determined by trypan blue exclusion assay. Induction of apoptosis was determined by nuclear condensation or fragmentation after Hoechst staining. Amount of apoptosis was quantified both by nuclear morphology and flow cytometry. Expression level of Bcl-2, and caspase-3 and PARP activations were assayed by Western blot. RESULTS: SKI 306X significantly prevented staurosporin-induced apoptosis. Among its three components, only Clematis mandshurica significantly decreased the amount of staurosporin-induced apoptosis. Although the level of Bcl-2 expression was decreased after staurosporin treatment, it was sustained after the combination treatment with Clematis mandshurica. Whereas staurosporin induced the degradation of 32 kDa caspase-3 precursor and the production of 85-kDa cleavage products of PARP in a time-dependent fashion, Clematis mandshurica treatment prevented those manifestations. CONCLUSIONS: Pharmacological efficacy of SKI 306X protecting osteoarthritis in part may result from the inhibition of apoptosis in chondrocytes by Clematis mandshurica.     

六味地黄丸对早期糖尿病肾病患者红细胞醛糖还原酶活性的影响

目的观察中药六味地黄丸对早期糖尿病肾病(DN)患者红细胞醛糖还原酶(AR)活性的抑制作用,以探讨六味地黄丸作为醛糖还原酶抑制剂防治DN的临床意义。方法选用我院诊断为早期DN且中医辨证为气阴两虚证的患者72例,按随机对照原则分为对照组(31例,常规治疗,即口服糖适平或注射胰岛素)和治疗组(41例,常规治疗加六味地黄丸),3个月为1个疗程,观察治疗前后DN的症状与体征;测定空腹血糖(FBG)、早餐后2小时血糖(2hPBG)、血胆固醇(TC)、甘油三酯(TG)、红细胞AR活性、尿白蛋白排泄率(UAER),血、尿β2-微球蛋白(β2-MG)的改善情况。结果(1)六味地黄丸可使DN患者的症状与体征改善;(2)六味地黄丸使红细胞AR活性受到抑制,明显低于对照组(P＜0．05);UAER,血β2-MG明显低于对照组(P＜0．05);(3)对血糖、血脂、平均动脉血压无明显影响(P＞0．05)。结论六味地黄丸可明显抑制早期DN红细胞AR活性,改善DN各项指标,有助于早期DN的治疗     

肾康注射液与苯那普利对人肾小球内皮细胞外基质抑制作用的对比研究

目的探讨肾康注射液与苯那普利对人肾小球内皮细胞外基质（ECM）不同成分的抑制作用强度。方法采用体外培养的人肾小球内皮细胞（EC）技术和血清药理学方法,观察具有降逆泄浊、益气活血功效的中药肾康注射液及苯那普利对ECM不同成分的影响。结果肾康注射液与苯那普利对ECM中纤维连接蛋白（FN）、层粘连蛋白（LN）和Ⅳ型胶原（ColⅣ）均具有一定的抑制作用,但在等效剂量浓度下,肾康注射液对ECM的抑制作用优于苯那普利。结论EC是肾康注射液发挥治疗作用的重要靶细胞之一,抑制ECM聚积可能是该方延缓肾小球硬化的重要作用机制之一。