高糖对体外培养大鼠肾小球系膜细胞VEGF和PEDF表达的影响

目的观察高糖环境对肾小球系膜细胞(HBZY-1)血管内皮生长因子(VEGF)和色素上皮衍生因子(PEDF)表达的影响,进一步探讨糖尿病肾病的发生、发展机制。方法体外常规培养HBZY-1,传代后分五组培养:对照组培养液葡萄糖浓度5.6 mmol/L,高糖a、b、c、d组培养液葡萄糖浓度分别为10、15、20、30 mmol/L;分别作用24、48 h后RT-PCR法测定各组细胞VEGF和PEDF mRNA表达水平;用ELISA法测定各组细胞培养液中VEGF和PEDF蛋白含量。结果与对照组相比,高糖各组细胞VEGF mRNA及VEGF蛋白的表达均升高,PEDF mRNA及PEDF蛋白表达均降低(P<0.05);高糖各组随葡萄糖浓度升高,VEGF表达升高、PEDF表达降低,呈浓度依赖性。结论高糖可导致肾小球系膜细胞VEGF和PEDF表达失衡,可能为糖尿病肾病微血管病变及肾小球硬化发生发展的机制之一     

高糖环境下VEGF与PEDF在GMC中表达及rAAV-AS基因的干预作用

目的 探讨高糖对大鼠肾小球系膜细胞(GMC)色素上皮细胞衍生因子(PEDF) 和血管内皮生长因子(VEGF)表达影响以及腺相关病毒介导血管抑素(rAAV-AS)基因的干预作用.方法 将培养的大鼠GMC株分为6组,高糖作为刺激因素,rAAV-AS 作为干预因素.分别设低糖组、高糖组、高糖加腺相关病毒(rAAV)组及高糖加rAAV-AS 治疗组.用免疫细胞化学及ELISA法测定各组系膜细胞PEDF 以及VEGF蛋白表达.结果 高糖可下调GMC中PEDF蛋白的表达,上调GMC中VEGF蛋白的表达.rAAV-AS可逆转高糖导致GMC的VEGF蛋白表达的增加及PEDF蛋白表达的降低.结论 rAAV-AS可以一定程度改善高糖诱导的VEGF和PEDF表达失衡而发挥对糖尿病肾病(DN)的保护作用.     

VEGF和色素上皮衍生因子的表达失衡对糖尿病肾病的影响

建立链脲佐菌素诱导的糖尿病大鼠模型,用免疫组化法观察肾小管间质血管内皮生长因子(VEGF)和色素上皮衍生因子(PEDF)的表达变化;体外培养大鼠肾成纤维细胞(NRK),用RT-PCR检测高糖对VEGF和PEDF mRNA表达的影响.结果 显示糖尿病大鼠肾脏组织VEGF表达增加,而PEDF表达下降.在NRK细胞,高糖呈时间、剂量依赖性地增加VEGF和抑制PEDF的mRNA表达.提示VEGF和PEDF的表达失衡可能在糖尿病肾病的发生发展中起一定作用.     

血管生成素1对高糖培养内皮细胞血管生成调控因子表达的影响研究

目的观察高糖培养人脐静脉内皮细胞(HUVECs)血管生成调控因子表达变化,探讨血管生成素1(Ang-1)对血管生成调控因子的影响。方法将HUVECs随机分为正常组(NC)、甘露醇组(MT)、高糖组(HG)和Ang-1组。加入不同处理因素后培养24、48h,采用免疫组织化学法及RT-PCR检测血管生成素2(Ang-2)、色素上皮衍生因子(PEDF)、血管内皮生成因子(VEGF)蛋白及mRNA表达水平。结果与NC组比较,24h时HG组Ang-2、VEGF的蛋白及mRNA表达均上调[(26.99±1.69)vs(23.22±1.51),P0.05;(37.32±1.39)vs (29.29±0.78),P0.05;(165.00±32.08)vs(108.00±9.64),P0.05;(185.33±14.44)vs(113.33±16.48),P0.05],且48h上调明显,PEDF两时点蛋白及mRNA表达下调[(30.72±1.32)vs(37.81±1.43),(26.73±0.86)vs(37.43±1.48),P0.05;(69.00±12.12)vs(129.33±19.74),(30.33±6.69)vs (122.67±20.88),P0.05]。与同时点HG组比较,Ang-1组的Ang-2、VEGF蛋白及mRNA降低,PEDF蛋白及mRNA升高(P0.05)。结论高糖培养HUVECs导致血管生成调控因子VEGF、Ang-2、PEDF表达异常。外源性给予Ang-1可缓解高糖导致血管生成调控因子的异常表达。     

葡萄糖对内皮祖细胞表达低氧诱导因子1α的影响

目的 观察葡萄糖对内皮祖细胞(EPC)表达低氧诱导因子1α(HIF-1α)和血管内皮生长因子(VEGF)的影响.方法 在EPC培养体系中加入不同浓度葡萄糖,分别在常氧(21%氧浓度)和低氧(1%氧浓度)条件下培养.检测各组HIF-1α及VEGF的基因和蛋白表达水平.结果 葡萄糖浓度相同,低氧组表达HIF-1α mRNA高于常氧组,VEGF的表达无统计学差异;氧浓度相同,10mmol/L葡萄糖组EPC表达HIF-1α mRNA高于其余两组,且随着葡萄糖浓度增加,VEGF表达逐渐下降.常氧浓度下测不到HIF-1α蛋白表达;低氧浓度下,10mmol/L葡萄糖组EPC表达HIF-1α最高.结论 高糖对低氧条件下的EPC具有毒性作用,能减弱其表达HIF-1α和VEGF.     

表观遗传修饰炎症相关基因在高血糖“代谢记忆”中的作用及机制

目的研究表观遗传修饰炎症相关基因在高血糖"代谢记忆"中的作用及相关机制。方法将Wistar大鼠分为正常对照组、高糖组和代谢记忆组,对比3组内皮细胞一氧化氮(NO)活性水平、单核细胞趋化蛋白(MCP-1)、细胞间黏附分子(ICAM-1)和白细胞介素(IL)-6 mRNA表达水平、活性氧(ROS)水平、内源性NO合酶(eNOS)基因表达和eNOS(Ser-1177)磷酸化水平。结果正常对照组NO含量显著高于高糖组和代谢记忆组,而MCP-1、ICAM-1和IL-6 mRNA表达则显著低于高糖组和代谢记忆组(P0.01);代谢记忆组NO含量显著高于高糖组,而MCP-1、ICAM-1和IL-6 mRNA表达则显著低于高糖组(P0.01)。正常对照组的超氧化物阴离子荧光探针(DHE)和活性氧簇细胞渗透性指示剂(CM-H2DCFDA)荧光强度显著低于高糖组和代谢记忆组(P0.01);代谢记忆组DHE和CM-H2DCFDA荧光强度显著低于高糖组(P0.01)。正常对照组eNOS mRNA和蛋白表达水平显著低于高糖组和代谢记忆组,而Ser-1177磷酸化水平显著高于高糖组和代谢记忆组(P0.01);代谢记忆组eNOS mRNA和蛋白表达水平显著低于高糖组,而Ser-1177磷酸化水平显著高于高糖组(P0.01)。结论早期高血糖可持续损伤血管内皮细胞舒张功能,进而令其功能紊乱,即使血糖水平恢复正常,仍然存在着持久的eNOS高表达,产生高血糖代谢记忆。     

大鼠脉络膜新生血管色素上皮衍生因子、血管内皮生长因子、血管生成素-2的表达

目的探讨大鼠脉络膜新生血管(DNV)组织色素上皮衍生因子(PEDF)和血管内皮生长因子(VEGF)、血管生成素-2(Ang-2)表达,探讨它们在CNV中的作用机制。方法应用氪激光光凝视网膜制备BN大鼠脉络膜新生血管(CNV)模型,随机分为6组,分别在光凝后1,3,7,14,21,28 d行荧光素眼底血管造影(FFA)后摘除眼球。应用实时荧光定量反转录PCR及免疫组化(SP)法检测各组大鼠光凝区和正常视网膜中PEDF、VEGF和Ang-2 mRNA和蛋白的表达,并对结果进行分析。结果光凝后3 d激光损伤区PEDF mRNA和蛋白表达程度最高,与其他各组比较差异显著(均P<0.05),其后表达逐渐下降;光凝后14 d CNV增殖区VEGF表达程度最高,与其他各组比较差异显著(均P<0.05);7 d时Ang-2表达程度最高,与其他各组比较差异显著(均P<0.05)。结论 PEDF表达不足可能是CNV形成和增殖一个主要原因,VEGF与PEDF表达失衡可能在CNV形成过程中起重要调控作用。     

蛋白激酶Cβ_2通过调控PPARα介导高糖诱导的人内皮细胞损伤

目的 研究蛋白激酶C(PKC)β_2、PPARα在高糖诱导入脐静脉内皮细胞(HUVECs)表达血管内皮生长因子(VEGF)、血管细胞黏附分子1(VCAM-1)mRNA中的作用及相互关系.方法 将培养的HUVECs分为以下8组:正常糖(NG,5 mmol/L D-葡萄糖)组、高糖(HG,25 mmol/L D-葡萄糖)组、渗透压对照(L,NG+20 mmol/L L-葡萄糖)组、正常糖卒载体转染(NN,NG+Ad5-null)组、高糖PKCβ_2转染(HB,HG+Ad5-PKCβ_2)组、高糖+非诺贝特(HF,HG+40 μmol/L非诺贝特)组、高糖PKCβ_2转染+非诺贝特(HBF,HB+40μmol/L非诺贝特)组,另以非诺贝特共孵育20 min作为HF20组,以上各组细胞均培养6 d.以RT-PCR 检测VEGF、VCAM-1 mRNA的表达水平,用Western印迹法测定PPARα蛋白表达,采用激光共聚焦检测PKCβ_2蛋白的表达和转位.结果 (1)HG组VEGF、VCAM-1 mRNA表达增加,分别为NG组的1.91倍和1.56倍(均P<0.05);HB组VEGF、VCAM-1 mRNA表达较HG组进一步增加,分别为NG组的2.59倍和2.07倍(均P<0.05).HF组VEGF、VCAM-1 mRNA表达则明显下调,分别为HG组的68%和74%(均P<0.05);与HG组相比,HF20组VEGF、VCAM-1 mRNA表达差异无统计学意义.(2)HG组PPARa蛋白表达较NG组减少了20%,HB组PPARα蛋白水平进一步下降,为HG组的78%,与HG组相比,HF组PPARα蛋白水平上调了13%(P<0.05).(3)HG诱导PKCβ_2核转位激活,定量分析示HG组浆/核荧光强度比值较NC组降低37%(P<0.05),HB组PKCβ_2核转位与HG组相比更加明显.结论 高糖通过诱导HUVECs PKCβ_2核转位,进而调控PPARα表达,增加VEGF、VCAM-1 mRNA的表达.     

色素上皮衍生因子对高糖环境下大鼠视网膜Müller细胞的影响

目的 探讨高糖环境下色素上皮衍生因子(PEDF)对大鼠视网膜Müller细胞的影响.方法将体外25 mmoL/L葡萄糖培养的大鼠Müller细胞以100 ng/ml PEDF或10 ng/ml白细胞介素-1β(IL-1β)孵育24 h,采用间接免疫荧光、Western印迹和实时定量PCR检测相关蛋白和mRNA表达,采用MTT检测Müller细胞活性.结果细胞免疫荧光组化、Western印迹和实时定量PCR检测显示在高糖环境下PEDF可以下调Müller细胞IL-1β蛋白和mRNA的表达,同样IL-1β也可以下调Müller细胞PEDF蛋白和mRNA的表达(P＜0.05);PEDF明显提高视网膜Müller细胞活性(0.48±0.09对0.64±0.17,P＜0.05).结论模拟糖尿病状态下,PEDF可以下调Müller细胞中IL-1β的表达,提高Müller细胞活性,可能对糖尿病炎症引起的病理改变起到一定抑制作用.     

腹膜透析液对大鼠腹膜间皮细胞血管内皮生长因子及其受体表达的影响

目的观察不同浓度葡萄糖腹膜透析液(dextrose)、艾考糊精腹膜透析液(icodextrin)对大鼠腹膜间皮细胞血管内皮生长因子(VEGF)及其受体VEGFR1、VEGFR2、sF lt-1合成的影响。方法分离、培养SD雄性大鼠腹膜间皮细胞(RPMCs),第二代细胞用于实验研究。实验分为5组,分别以不同腹膜透析液[1.50%dextrose(低糖组)、2.50%dextrose(中糖组)、4.25%dextrose(高糖组)、7.50%icodextrin组(糊精组)]进行刺激培养,无血清DMEM为阴性对照(对照组)。RT-PCR法检测VEGF及其受体VEGFR1、VEGFR2、sFlt-1 mRNA表达,Western印迹法检测RPMCs的VEGFR1、VEGFR2蛋白表达;酶联免疫吸附试验(ELISA)检测细胞培养上清中VEGF及sFlt-1表达。结果各组均见VEGFR1、VEGFR2、sFlt-1 mRNA和蛋白表达。在核酸和蛋白水平,中糖组、高糖组VEGFR1表达明显高于低糖组、糊精组及对照组(P0.01,P0.05);中糖组、高糖组VEGFR2表达显著低于低糖组、糊精组及对照组(均P0.01);低糖组VEGFR2蛋白表达显著低于对照组(P0.01)。在核酸水平,中糖组、高糖组VEGF、sFlt-1表达明显高于对照组(P0.01);RPMCs培养上清液中,VEGF、sFlt-1蛋白表达强弱趋势与RT-PCR一致。结论正常培养的RPMCs表达VEGFR1、VEGFR2及sFlt-1。腹膜透析液,尤其是高浓度dextrose,能明显上调VEGF、VEGFR1及sFlt-1的表达,但下调VEGFR2的表达,提示其可通过调节VEGF受体从而调节VEGF系统,这可能是导致长期腹膜透析过程中腹膜血管增生的机制之一。icodextrin的生物相容性可能优于高浓度dextrose。     

Pigment-epithelium-derived factor (PEDF) inhibits angiotensin-II-induced vascular endothelial growth factor (VEGF) expression in MOLT-3 T cells through anti-oxidative properties

Angiotensin II (Ang II), the dominant effector of the renin-angiotensin system, elicits numerous inflammatory-proliferative responses, thereby being involved in angiogenesis. T cells play an important role in angiogenesis as well by delivering vascular endothelial growth factor (VEGF) to inflammatory sites. Since we have previously shown that pigment-epithelium-derived factor (PEDF) blocks the Ang II signaling in endothelial cells, we studied here whether PEDF could inhibit the Ang-II-induced VEGF expression in MOLT-3 T and examined the potential mechanism of PEDF inhibitory effects. Ang II significantly up-regulated VEGF mRNA levels in MOLT-3 T cells, which was inhibited by PEDF or olmesartan, an Ang II type 1 receptor blocker. PEDF blocked the Ang-II-induced reactive oxygen species (ROS) generation in MOLT-3 T cells. Furthermore, H(2)O(2) was found to up-regulate VEGF mRNA levels in MOLT-3 T cells in a dose-dependent manner. These results demonstrate that PEDF could inhibit the Ang-II-induced VEGF expression in MOLT-3 T cells via suppression of ROS generation. Blockade by PEDF of VEGF expression in T cells may become a novel therapeutic target for pathological angiogenesis.     

Pigment epithelium-derived factor inhibits leptin-induced angiogenesis by suppressing vascular endothelial growth factor gene expression through anti-oxidative properties

Leptin, a circulating hormone secreted mainly from adipose tissues, is involved in the control of body weight. Recently, leptin was found to be an angiogenic factor, and its vitreous levels are associated with angiogenic eye diseases such as proliferative diabetic retinopathy. However, the molecular mechanism for leptin-elicited angiogenesis remains to be elucidated. Pigment epithelium-derived factor (PEDF) has been shown to be the most potent natural inhibitor of angiogenesis in the mammalian eye, and its levels in the vitreous were decreased in angiogenic eye diseases. In this study, we investigated whether and how PEDF could inhibit the leptin-induced DNA synthesis in microvascular endothelial cells (EC), a key step of angiogenesis. Leptin significantly increased intracellular reactive oxygen species (ROS) generation in microvascular EC. PEDF was found to inhibit the leptin-induced ROS generation in EC. An anti-oxidant, N-acetylcysteine, or PEDF completely prevented the leptin-induced upregulation of vascular endothelial growth factor (VEGF) mRNA levels as well as any increase in DNA synthesis in microvascular EC. Polyclonal antibodies against human VEGF were also found to completely inhibit DNA synthesis in leptin-exposed EC. The present study suggests that leptin could elicit angiogenesis through autocrine VEGF production via intracellular ROS generation. PEDF may block the angiogenic effects of leptin through its anti-oxidative properties.     

Pigment epithelium-derived factor inhibits oxidative stress-induced apoptosis and dysfunction of cultured retinal pericytes

Pigment epithelium-derived factor (PEDF) is a potent inhibitor of angiogenesis in the mammalian eye, suggesting that loss of PEDF is implicated in the pathogenesis of proliferative diabetic retinopathy. However, a role for PEDF in early diabetic retinopathy remains to be elucidated. Since oxidative stress is thought to be involved in pericyte loss and dysfunction, one of the changes characteristic of early diabetic retinopathy, we investigated whether and how PEDF could protect cultured retinal pericyte against oxidative stress injury. High glucose (30 mM) increased intracellular reactive oxygen species (ROS) generation in pericytes, which was completely blocked by PEDF. High glucose or H2O2 was found to induce growth retardation and apoptotic cell death of pericytes. PEDF completely restored these cytopathic effects on pericytes. An increased ratio of bax to bcl-2 mRNA level with subsequent activation of caspase-3 was observed in high-glucose- or H2O2-exposed pericytes, which was also completely prevented by PEDF. PEDF significantly increased glutathione peroxidase (GPx) mRNA levels and activity in pericytes. Further, PEDF was found to completely inhibit high-glucose- or H2O2-induced increase in a mRNA ratio of angiopoietin-2 to angiopoietin-1 and up-regulation of VEGF mRNA levels in pericytes. PEDF mRNA levels themselves were down-regulated in high-glucose- or H2O2-exposed pericytes. These results demonstrate that PEDF protects against high-glucose- or H2O2-induced pericyte apoptosis and dysfunction through its anti-oxidative properties via GPx induction. Our present study suggests that substitution of PEDF proteins might be a promising therapeutic strategy for treatment of patients with early diabetic retinopathy.     

Pigment epithelium-derived factor inhibits TNF-alpha-induced interleukin-6 expression in endothelial cells by suppressing NADPH oxidase-mediated reactive oxygen species generation

Pigment epithelium-derived factor (PEDF) has recently been shown to be involved in the pathogenesis of proliferative diabetic retinopathy. Atherosclerosis is an inflammatory-fibroproliferative disease as well. Oxidative stress plays a major role in retinopathy and atherosclerosis. Accordingly, we investigated effects of PEDF on reactive oxygen species (ROS) generation, NF-kappaB activation and interleukin (IL)-6 expression in TNF-alpha-exposed HUVEC. TNF-alpha significantly increased intracellular ROS generation, which was completely blocked by PEDF or diphenylene iodonium, an inhibitor of NADPH oxidase. Further, PEDF completely prevented the TNF-alpha-induced increase in NADPH oxidase activity. PEDF or an antioxidant, N-acetylcysteine, significantly inhibited the TNF-alpha-induced NF-kappaB activation. PEDF inhibited TNF-alpha-induced expression of IL-6 at both mRNA and protein levels. Moreover, TNF-alpha downregulated PEDF mRNA levels. Ligand blot analysis revealed that HUVEC possessed a membrane protein with binding affinity for PEDF. The results demonstrated that PEDF inhibited TNF-alpha-induced NF-kappaB activation and subsequent IL-6 overexpression in HUVEC by suppressing NADPH oxidase-mediated ROS generation. Our present study suggests that PEDF may play an important role in the development and progression of atherosclerosis.     

Activation of nicotinamide adenine dinucleotide phosphate (reduced form) oxidase by advanced glycation end products links oxidative stress to altered retinal vascular endothelial growth factor expression

Increasing evidence indicates that advanced glycation end products (AGEs) promote retinal alterations through oxidative stress. However, the pathways involved in AGE-induced generation of reactive oxygen species (ROS) in retinal cells are poorly defined. In the present study, we investigated the role of nicotinamide adenine dinucleotide phosphate (reduced form) (NADPH) oxidase in AGE-induced ROS intracellular generation and vascular endothelial growth factor (VEGF) expression in bovine retinal endothelial cells (BRECs). Incubation of BRECs with 100 microg/mL AGEs increased ROS generation and VEGF expression in these cells. Treatment of the cells with the NADPH oxidase inhibitors, apocynin and diphenylene iodonium, inhibited these effects. In retinal endothelial cells exposed to AGEs, translocation of protein kinase C (PKC)-beta2 and p47phox was observed. Inhibition of PKC by treatment of the cells with calphostin C, GF10923X, and LY379196 totally suppressed AGE-mediated p47phox translocation and ROS generation. Incubation of BRECs with gliclazide inhibited AGE-induced PKC-beta2 and p47phox translocation and totally abrogated AGE-mediated ROS generation and VEGF expression. Overall, these results demonstrate that AGEs induce intracellular ROS generation and VEGF expression in retinal endothelial cells through a PKC-dependent activation of NADPH oxidase. Inhibition of retinal NADPH oxidase expression and ROS generated by this system provides a new potential mechanism by which gliclazide may affect retinal VEGF expression and exert a beneficial effect on diabetic retinopathy.     

色素上皮衍生因子抑制高糖诱导的人肾小球系膜细胞转化生长因子β1、纤连蛋白和Ⅳ型胶原的表达

随着葡萄糖浓度的增加,人肾小球系膜细胞转化生长因子β1(TGF-β1)、纤连蛋白(FN)和Ⅳ型胶原(ColⅣ)的表达显著增加(P＜0.01).在30 mmol/L的高糖条件下,40～160 nmol/L的色素上皮衍生因子(PEDF)显著抑制TGF-β1表达,呈剂量依赖性(P＜0.01);5～40 nmol/L的PEDF也剂量依赖性地抑制FN和ColⅣ的表达(P＜0.01),提示PEDF可能通过其抗纤维化的作用延缓糖尿病肾病的进程.

Abstract：

The expressions of transforming growth factor β1 (TGF-β1), fibronectin (FN), and collagen Ⅳ in human mesangial cells were augmented with increased concentrations of glucose.Pigment epithelium-derived factor (PEDF) at concentrations of 40-160 nmol/L significantly inhibited TGF-β1 expression in a dose-dependent manner(P＜0.01).PEDF at concentrations of 5-40 nmol/L significantly decreased FN and collagen Ⅳ expressions in a dose-dependent manner(P＜0.01) ,suggesting that PEDF may play a salutary role in diabetic nephropathy by its antifibrogenic activity.     

冬虫夏草对高糖诱导的大鼠肾小球系膜细胞色素上皮衍生因子和血管内皮生长因子表达的影响

目的观察冬虫夏草对高糖诱导的大鼠肾小球系膜细胞色素上皮衍生因子（PEDF）和血管内皮生长因子（VEGF）表达的影响,探讨其肾脏保护的作用机制。方法35只8周龄清洁级雄性sD大鼠,体重260～280g,根据体重按随机数字表法分为正常血清组（n=18）、低剂量虫草提取液喂养组（5g·kg^-1·d^-1,n=8）和高剂量虫草提取液喂养组（10g·kg^-1·d^-1,n=9）,采用生理盐水及不同剂量虫草提取液灌胃8d后,分离静脉血提取对照血清及不同浓度的冬虫夏草含中药血清。体外培养HBZY-1大鼠肾小球系膜细胞,之后分别加入正常血糖正常血清[正常血糖组（NC组）：5．6mmol／L葡萄糖＋10％胎牛血清（FBS）＋1％正常大鼠血清]、高血糖正常血清[高血糖组（HG组）：30．0mmol／L葡萄糖＋10％FBS＋1％正常大鼠血清]、低剂量虫草含药血清[低药组（LD）：30．0mmol／L葡萄糖＋10％FBS＋1％低剂量虫草含药血清]和高剂量虫草含药血清[高药组（HD）：30．0mmol／L葡萄糖＋10％FBS＋1％高剂量虫草含药血清]进行培养,采用逆转录．聚合酶链反应（RT—PCR）测定各组细胞PEDF及VEGFmRNA的表达,酶联免疫吸附试验（ELISA）测定各组细胞上清液中PEDF及VEGF的水平。组间资料比较采用单因素方差分析,样本间资料比较采用t检验。结果与NC组相比,HG组PEDFmRNA表达明显下降（24h：0．375±0．011比0．507±0．008,t=16．828,P〈0．0l;72h：0．376±0．014比0．515±0．010,t=14．034,P〈0．01）,VEGFmRNA表达明显上升（24h：1．005±0．011比0．864±0．012,t=14．963,P〈0．01;72h：1．168±0．012比0．873±0．011,t=31．417,P〈0．01）;LD组和HD组PEDFmRNA表达较HG组显著上调（24h：0．505±0．018、0．639±0．012比0．375±0．011,F=354．719,P〈0．01;72h：0．612±0．023、0．700±0．019比0．376±0．014,F=97．82,P〈0．01）,VEGFmRNA表达明显下降（24h：0．838±0．014、0．767±0．017比1．005±0．011．F=79．55,P〈0．01：72h：0．923±0．016、0．784±0．012比1．168±0．012,F=244．28,P〈0．01）,且HD组疗效更明显。经对照血清及CS血清培养24、72h后,与NC组相比,HG组PDEF水平明显降低[24h：（267±8）比（303±10）ng／L,t=6．493,P〈0．01;72h：（265±27）比（338±42）ng／L,t=3．259,P〈0．01],VEGF明显升高[24h：（128±3）比（113±8）ng／L,t=3．737,P〈0．01;72h：（144±7）比（125±7）ng／L,t=4．215,P〈0．01;与HG组相比,LD组和HD组PEDF明显升高[24h：（297±12）、（296±26）比（267±8）ng／E,F=5．427,P〈0．01;72h：（323±20）、（332±33）比（265±27）ng／L,F=5．690,P〈0．01],VEGF明显降低[24h：（106±6）、（109±11）比（128±3）ng／L,F=5．738,P〈0．01;72h：（124±9）、（125±7）比（144±7）ng／L,F=7．878,P〈0．01]。结论冬虫夏草可能通过降低肾小球系膜细胞VEGF的表达,升高PEDF的表达,对肾脏起到保护作用。