复方黄芪首乌合剂对代谢综合征（MS）大鼠早期肾损伤的保护作用及分子机制研究
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摘要
代谢综合征(Metabolic syndrome, MS)是以多种代谢性疾病合并出现(包括中心性肥胖、糖尿病或糖调节受损、高血压、脂代谢紊乱等)为特点,以胰岛素抵抗为共同病理生理基础的一组临床症候群。近年来随着经济的发展和生活方式的改变,其发病率逐年增高,已成为研究的热点。
MS中的任何单一因素均可引起和加重肾损害,无论是肥胖,还是糖尿病、高血脂和高血压。在早期阶段表现为高滤过、高灌注和肾体积肥大,并可见微量白蛋白尿；随着病情的进一步发展分别表现为肥胖相关性局灶节段性肾小球硬化、糖尿病肾病和高血压肾病(中期)；但最后均发展为终末期肾衰竭,即尿毒症。因此,要防治MS引起的肾损害,应尽量在发病早期进行干预,阻断肾损害的进一步进展的通路。
西医对MS治疗目前尚无统一标准,治疗目标集中在预防糖尿病和心血管事件发生。需要长期给予多种药物,分别进行调脂、改善胰岛素抵抗、降压和减肥等治疗。但是,多种西药的长期联合应用容易导致药物的相互拮抗作用和不良反应的发生。因此,人们开始重视中医药防治MS的研究。一方面,中医药作为一种自然疗法,可以与行为干预(饮食与运动)相结合,更易于被患者接受,另外,中药具有多途径多靶点的整体调节模式,同时制剂相对价廉。
中医认为过食和少动是MS的两大主因,从而造成食、气、血、火、痰、湿六郁作用于脾胃而酿成痰、瘀、浊、脂等病理产物。复方黄芪首乌合剂(HQSW)为纯中药制剂,由黄芪、首乌、丹参、山楂、决明子、红曲、虎杖七味药组成,方中黄芪、首乌益气补肾；丹参、山楂活血化瘀；决明子、红曲消食化浊；虎杖解毒利湿,以上诸药融于一方,共行健脾益肾,化浊祛瘀之功。
综上,在下文中我们将分四部分对MS大鼠的糖脂代谢、氧化应激反应、内皮功能和足细胞变化及HQSW的干预作用进行探讨：①HQSW对代谢综合征大鼠糖脂代谢和肾功能的影响；②HQSW对MS大鼠肾组织(?)AngⅡ和FFA分泌及氧化应激反应的影响；③HQSW对MS大鼠一氧化氮和内皮素-1分泌及其基因表达的调节作用；④HQSWMS大鼠足细胞损伤的保护及Podocin和Nephrin表达的调节作用。
第-部分复方黄芪首乌合剂对代谢综合征大鼠模型的糖脂代谢和肾功能的影响
目的观察HQSW对实验性MS大鼠血糖、血脂、血压和肾功能的改善作用。方法采用高糖高脂饲料喂养建立实验性MS大鼠模型,以复方黄芪首乌合剂(低剂量组、中剂量组和高剂量组)灌胃治疗,8周末,采血检测血脂、血糖和肾功能,光镜下观察肾脏病理改变。
结果实验8周末,模型组大鼠体重显著增加,血糖、TCH、TG和LDL显著升高(P<0.05),而高密度脂蛋白明显下降(P<0.05),同时胰岛素敏感指数显著降低(P<0.05),符合MS诊断指征。经过中药干预治疗后,实验大鼠体重比模型组明显下降,与西药(二甲双胍)对照组结果相似(P<0.05)；HQSW能够明显降低MS大鼠血清TCH、TG和LDL(P<0.05),使实验大鼠的HDL-C显著升高(P<0.05)；HQSW可以改善MS大鼠的糖代谢紊乱,使实验大鼠血清FPG、PBG、Fins显著减低,ISI显著升高；HQSW虽然对实验大鼠血肌酐、尿素氮和血尿酸水平明显下降(P>0.05)；肾脏病理显示,中药治疗组大鼠肾病病理损害明显减轻。
结论采用高糖高脂喂饲8周可成功建立实验性MS大鼠模型。HQSW能提高MS大鼠胰岛素敏感性,调节脂代谢紊乱,降低血糖,对MS大鼠早期肾脏病理损害具有保护作用。
第二部分复方黄芪首乌合剂对MS大鼠肾组织AngⅡ、FFA分泌及氧化应激反应的作用
目的探讨复方黄芪首乌合剂(HQSW)对实验性MS大鼠肾组织血管紧张素Ⅱ(AngⅡ)和游离脂肪酸(FFA)分泌及氧化应激反应的影响。
方法采用高糖高脂饲料喂养建立实验性MS大鼠模型,以HQSW(低剂量组、中剂量组和高剂量组)灌胃治疗,8周末,比色法测定实验鼠血浆和肾皮质A丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性,放射免疫法测定血浆和肾皮质FFA和AngⅡ浓度变化。
结果MS大鼠血浆和肾组织SOD活力显著下降(P<0.01),而MDA、FFA和AngⅡ水平显著上升(P<0.01)。经中药HQSW干预后,血浆和肾组织SOD活力显著上升(P<0.05)；同时MDA、FFA和AngⅡ水平显著下降(P<0.05)。
结论MS大鼠FFA和AngⅡ的分泌异常增多,同时出现异常氧化应激反应,通过HQSW的干预作用,上述异常状况得到极大地改善,这可能是其防治MS大鼠肾损伤的关键机制之一。
第三部分复方黄芪首乌合剂对MS大鼠一氧化氮和内皮素-1分泌及其基因表达的调节作用
目的观察HQSW对实验性MS大鼠血压的影响及其对一氧化氮(NO)和内皮素-1(ET-1)平衡的调节作用。
方法采用高糖高脂饲料喂养建立实验性MS大鼠模型,以HQSW(低剂量组、中剂量组和高剂量组)灌胃治疗,8周末,采用多功能生理仪测定实验鼠SBP、DBP、MAP,比色法测定实验鼠血浆和肾皮质匀浆中NO含量和一氧化氮合酶(NOS)活性,放射免疫法测定血浆和肾皮质匀浆中ET-1的含量,逆转录-聚合酶链反应(RT-PCR)检测肾皮质ET-1、NOS的mRNA表达。
结果MS大鼠SBP、DBP、MAP均有不同程度增高,血浆和肾组织NO、NOS、和肾脏NOS基因表达水平显著下降(P<0.05),同时血浆和肾组织中ET-1和肾脏ET-1mRNA表达水平显著上升(P<0.05),经中药HQSW干预治疗后,HQSW可以改善MS大鼠的高血压状况,使实验大鼠SBP、DBP、MAP均有不同程度减低；血浆和肾皮质匀浆中NO、NOS和肾组织NOSmRNA表达上调(P<0.05),相反,血浆和肾皮质ET-1浓度和肾组织ET-1mRNA表达下调(P<0.05)。
结论MS大鼠发生NO/ET-1平衡的紊乱,从而导致血压增高。HQSW可以改善MS大鼠的高血压状况,使实验大鼠SBP、DBP、MAP均有不同程度减低,它依赖于对大鼠的ET-1/NO的动态平衡具有调节作用,这可能是其发挥防治早期MS肾损伤的主要机制之一。
第四部分复方黄芪首乌合剂MS大鼠足细胞的保护及Podocin和Nephrin表达的调控作用
目的观察HQSW对实验性代谢综合征大鼠早期足细胞损伤的保护作用并探索其作用机制。
方法用高糖高脂饲料喂养建立实验性代谢综合征大鼠模型,以HQSW(低剂量组、中剂量组和高剂量组)灌胃治疗。8周后,比色法测定实验大鼠24小时尿蛋白(24hPro)；免疫比浊法测定尿微量白蛋白(MA)和尿微量白蛋白与尿肌酐比值(albumin to creatinine ratio, ACR);免疫组织化学法检测肾组织Nephrin和Podocin的蛋白表达；逆转录-聚合酶链反应(RT-PCR)检测肾皮质Nephrin和Podocin mRNA的表达；电镜下观察肾脏基底膜(GBM)和足细胞的变化。
结果MS大鼠24hPro.MA和ACR较正常对照组明显增加(P<0.01),肾小球基底膜显著增厚,足细胞与对照组大鼠比较明显减少,足突排列紊乱,足突融合；肾小球Podocin和Nephrin的蛋白和基因表达明显低于正常对照组(P<0.05)。经HQSW干预治疗8周后,24hPro、MA、MA/UCr明显减少(P<0.05)；肾小球基底膜病变减轻,细胞外基质增多减轻,足细胞数目增多,足细胞融合减轻；肾脏皮质Podocin和Nephrin的蛋白与基因表达分别较模型组最显著增加(P<0.05)。
结论MS大鼠肾脏足细胞发生明显的病理损害,同时伴随着Nephrin和Podocin的表达异常。HQSW能防治MS大鼠的足细胞损害,修复肾组织Nephrin和Podocin的蛋白和基因的异常表达,这可能是其发挥防治MS早期肾损伤的主要机制之一。
Part I The effects of the mixture of Radix Astragalus and Polygonum mulflorum on glucose and lipid metabolism and renal function in rats with MS
Objective To investigate the effect of mixture Radix Astragalus and Polygonum mulflorum on glucose and lipid metabolism and renal function in rats with metabolic syndrome.
Methods To establish experimental metabolic syndrome rat model through a high-sugar and high-fat diet, Animals were administered with the mixture of Radix Astragalus and Polygonum mulflorum (low dose group, middle dose group and high dose group). At the8th week, the serum were used to detect TG、TCH、Scr、Bun、 UA and Glu with biochemical analyzer, the pathologic change of kidney was observed by light microscope
Results At the end of8th week,In MS model group, there is a significant increase in body weight, serum glucose, cholesterol, triglycerides and low density lipoprotein, and accompany with the significant decrease of high-density lipoprotein and insulin sensitivity index, those satisfied the diagnostic indications of MS. The body weight of rats treated with the mixture of Radix Astragalus and Polygonum mulflorum was decreased significantly compared with model group, and there is similar results in rats treated with Metformin(P＜0.05); Radix Astragalus and Polygonum mulflorum mixture can significantly reduce serum levels of TCH, TG and LDL (P＜0.05), at the same time the serum level of HDL-C increased significantly (P＜0.05); there is significant change on Scr、BUN and UA in experimental rat treated with the mixture of Radix Astragalus and Polygonum mulflorum (P<0.05), the pathologic change was improved after treatment.
Conclusion:The model of metabolic syndrome in rats can be successfully replicated through High-sugar and high-fat diet. The mixture of Radix Astragalus and Polygonum mulflorum can improve insulin sensitivity, and regulate lipid metabolism, reduce blood glucose, and protect renal function. Above all it can improve the pathologic change in MS rats.
Part II The effects of the mixture of Radix Astragalus and Polygonum mulflorum on renal secretion of AngⅡ and FFA and oxidative stress in rats with MS
Objective To investigate the effect of the mixture of Radix Astragalus and Polygonum mulflorum on renal secretion of angiotensin Ⅱ (AngⅡ) and free fatty acids (FFA) and oxidative stress in rats with metabolic syndrome.
Methods To establish experimental metabolic syndrome rat model through a high-sugar and high-fat diet, Animals were administered with the mixture of Radix Astragalus and Polygonum mulflorum (low dose group, middle dose group and high dose group). At the end of8th week, the level of Super Oxygen Dehydrogenises (SOD) and Methane Dicarboxylic Aldehyde (MDA) in the serum and renal cortex were determined by colorimetric method, the level of AngⅡ and FFA in the serum and renal cortex were detected by radioimmunoassay.
Results The enzyme activity of SOD in the serum and renal cortex was decreased significantly in rats with MS (P＜0.01), on the contrary, the level of MDA、AngⅡ and FFA in the serum and renal cortex were increased significantly (P＜0.01). After treated with the mixture of Radix Astragalus and Polygonum mulflorum, The enzyme activity of SOD in the serum and renal cortex was increased significantly (P＜0.05), At the same time, The level of MDA、AngⅡ and FFA in the serum and renal cortex were decreased significantly (P＜0.05)
Conclusion The abnormal secretion of FFA and AngⅡ were occurred in rats with MS, at the same time, the oxidative stress response was being aggravated. After treated with the mixture of Radix Astragalus and Polygonum mulflorum, the above abnormal change was improved, that might be its prevention and treatment mechanism of early renal injury in rats with MS.
Part Ⅲ The regulation of mixture of Radix Astragalus and Polygonum mulflorum on balance of NO and ET-1in rats with MS
Objective To investigate the effect on blood pressure and regulation of the mixture of Radix Astragalus and Polygonum mulflorum on the balance of Nitric Oxide (NO) and endothelin-1(ET-1) in rats with metabolic syndrome.
Methods To establish experimental metabolic syndrome rat model through a high-sugar and high-fat diet, Animals were administered with Radix Astragalus compound mixture (low dose group, middle dose group and high dose group) for8weeks. Then, the level of SBP、DBP、MAP were detected with multifunction physiological monitor, The level of NO and the enzyme activity of NOS in the serum and renal cortex were determined by colorimetric method, he level of ET-1in the serum and renal cortex were detected by radioimmunoassay, The mRNA expressions of Renal ET-1and NOS were detected through reverse transcription polymerase chain reaction(RT-PCR).
Results The level of SBP、DBP、MAP in MS rats was increased significantly compared with normal rats (P＜0.05), The enzyme activity of NOS and the level of NO in the serum and renal cortex and the renal gene expression of NOS was decreased significantly in rats with MS (P＜0.05), On the contrary, the level of ET-1in the serum and renal cortex and the renal gene expression of ET-1was increased significantly (P＜0.05). After treated with the mixture of Radix Astragalus and Polygonum mulflorum, The level of SBP、DBP、MAP in MS rats was decreased significantly compared with normal rats (P＜0.05),The enzyme activity of NOS and the level of NO in the serum and renal cortex and the renal gene expression of NOS was increased significantly (P＜0.05)(P＜0.05), At the same time, the level of ET-1in the serum and renal cortex and the renal gene expression of ET-1was decreased significantly (P＜0.05)
Conclusion The balance of NO/ET-1was destroyed in rats with MS, and then the abnormal level of SBP、DBP、MAP were detected, The mixture of Radix Astragalus and Polygonum mulflorum can lower the blood pression in MS rats through to regulate the balance of NO/ET-1in MS rats, this might play key role in protection the early renal injury of MS rats.
Part IV The effects of the mixture of Radix Astragalus and Polygonum mulflorum on renal expression of Podocin and Nephrin in rats with MS
Objective To investigate the effects and mechanism of the mixture of Radix Astragalus and Polygonum mulflorum on early injury of renal podocyte in rats with metabolic syndrome.
Methods To establish experimental metabolic syndrome rat model through a high-sugar and high-fat diet, Animals were administered with Radix Astragalus compound mixture (low dose group, middle dose group and high dose group) for8weeks. Then, the level of24hPro was detected by colorimetric method; the level of MA and ACR were detected by lmmunoturbidimetry; the protein expression of Nephrin and Podocin in renal cortex was determined by immunohistochemistry; the gene expression of Nephrin and Podocin in renal cortex was determined by RT-PCR; The pathological change of Glomerular Basement Membrane(GBM) and podocyte was observed by electron microscope.
Results The level of24hPro、MA and ACR were increased significantly in rats with MS compared with normal rats(P＜0.05); Under electronic microscopic examination, fusion of foot process, disruption of GBM and enlargement of infiltration pores, thickness of GBM were observed; the protein and gene expression of Podocin and Nephrin in renal cortex was decreased significantly compared with normal rats(P＜0.05). After8weeks treatment with the mixture of Radix Astragalus and Polygonum mulflorum, The level of24hPro、MA and ACR were decreased significantly in rats with MS compared with normal rats(P＜0.05); the pathologic change of GBM and podocte was reduced significantly compared with MS rats; the protein and gene expression of Podocin and Nephrin in renal cortex was increased significantly compared with normal rats(P＜0.05).
Conclusion The pathological change of podocyte was observed in rats with MS, accompanied by the abnormal protein and gene expression of Nephrin and Podocin in renal cortex. After the treatment of the mixture of Radix Astragalus and Polygonum mulflorum, the abnormal condition was improved, and it might play a key role in protection the early renal injury of MS rats.
MS中的任何单一因素均可引起和加重肾损害,无论是肥胖,还是糖尿病、高血脂和高血压。在早期阶段表现为高滤过、高灌注和肾体积肥大,并可见微量白蛋白尿；随着病情的进一步发展分别表现为肥胖相关性局灶节段性肾小球硬化、糖尿病肾病和高血压肾病(中期)；但最后均发展为终末期肾衰竭,即尿毒症。因此,要防治MS引起的肾损害,应尽量在发病早期进行干预,阻断肾损害的进一步进展的通路。
西医对MS治疗目前尚无统一标准,治疗目标集中在预防糖尿病和心血管事件发生。需要长期给予多种药物,分别进行调脂、改善胰岛素抵抗、降压和减肥等治疗。但是,多种西药的长期联合应用容易导致药物的相互拮抗作用和不良反应的发生。因此,人们开始重视中医药防治MS的研究。一方面,中医药作为一种自然疗法,可以与行为干预(饮食与运动)相结合,更易于被患者接受,另外,中药具有多途径多靶点的整体调节模式,同时制剂相对价廉。
中医认为过食和少动是MS的两大主因,从而造成食、气、血、火、痰、湿六郁作用于脾胃而酿成痰、瘀、浊、脂等病理产物。复方黄芪首乌合剂(HQSW)为纯中药制剂,由黄芪、首乌、丹参、山楂、决明子、红曲、虎杖七味药组成,方中黄芪、首乌益气补肾；丹参、山楂活血化瘀；决明子、红曲消食化浊；虎杖解毒利湿,以上诸药融于一方,共行健脾益肾,化浊祛瘀之功。
综上,在下文中我们将分四部分对MS大鼠的糖脂代谢、氧化应激反应、内皮功能和足细胞变化及HQSW的干预作用进行探讨：①HQSW对代谢综合征大鼠糖脂代谢和肾功能的影响；②HQSW对MS大鼠肾组织(?)AngⅡ和FFA分泌及氧化应激反应的影响；③HQSW对MS大鼠一氧化氮和内皮素-1分泌及其基因表达的调节作用；④HQSWMS大鼠足细胞损伤的保护及Podocin和Nephrin表达的调节作用。
第-部分复方黄芪首乌合剂对代谢综合征大鼠模型的糖脂代谢和肾功能的影响
目的观察HQSW对实验性MS大鼠血糖、血脂、血压和肾功能的改善作用。方法采用高糖高脂饲料喂养建立实验性MS大鼠模型,以复方黄芪首乌合剂(低剂量组、中剂量组和高剂量组)灌胃治疗,8周末,采血检测血脂、血糖和肾功能,光镜下观察肾脏病理改变。
结果实验8周末,模型组大鼠体重显著增加,血糖、TCH、TG和LDL显著升高(P<0.05),而高密度脂蛋白明显下降(P<0.05),同时胰岛素敏感指数显著降低(P<0.05),符合MS诊断指征。经过中药干预治疗后,实验大鼠体重比模型组明显下降,与西药(二甲双胍)对照组结果相似(P<0.05)；HQSW能够明显降低MS大鼠血清TCH、TG和LDL(P<0.05),使实验大鼠的HDL-C显著升高(P<0.05)；HQSW可以改善MS大鼠的糖代谢紊乱,使实验大鼠血清FPG、PBG、Fins显著减低,ISI显著升高；HQSW虽然对实验大鼠血肌酐、尿素氮和血尿酸水平明显下降(P>0.05)；肾脏病理显示,中药治疗组大鼠肾病病理损害明显减轻。
结论采用高糖高脂喂饲8周可成功建立实验性MS大鼠模型。HQSW能提高MS大鼠胰岛素敏感性,调节脂代谢紊乱,降低血糖,对MS大鼠早期肾脏病理损害具有保护作用。
第二部分复方黄芪首乌合剂对MS大鼠肾组织AngⅡ、FFA分泌及氧化应激反应的作用
目的探讨复方黄芪首乌合剂(HQSW)对实验性MS大鼠肾组织血管紧张素Ⅱ(AngⅡ)和游离脂肪酸(FFA)分泌及氧化应激反应的影响。
方法采用高糖高脂饲料喂养建立实验性MS大鼠模型,以HQSW(低剂量组、中剂量组和高剂量组)灌胃治疗,8周末,比色法测定实验鼠血浆和肾皮质A丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性,放射免疫法测定血浆和肾皮质FFA和AngⅡ浓度变化。
结果MS大鼠血浆和肾组织SOD活力显著下降(P<0.01),而MDA、FFA和AngⅡ水平显著上升(P<0.01)。经中药HQSW干预后,血浆和肾组织SOD活力显著上升(P<0.05)；同时MDA、FFA和AngⅡ水平显著下降(P<0.05)。
结论MS大鼠FFA和AngⅡ的分泌异常增多,同时出现异常氧化应激反应,通过HQSW的干预作用,上述异常状况得到极大地改善,这可能是其防治MS大鼠肾损伤的关键机制之一。
第三部分复方黄芪首乌合剂对MS大鼠一氧化氮和内皮素-1分泌及其基因表达的调节作用
目的观察HQSW对实验性MS大鼠血压的影响及其对一氧化氮(NO)和内皮素-1(ET-1)平衡的调节作用。
方法采用高糖高脂饲料喂养建立实验性MS大鼠模型,以HQSW(低剂量组、中剂量组和高剂量组)灌胃治疗,8周末,采用多功能生理仪测定实验鼠SBP、DBP、MAP,比色法测定实验鼠血浆和肾皮质匀浆中NO含量和一氧化氮合酶(NOS)活性,放射免疫法测定血浆和肾皮质匀浆中ET-1的含量,逆转录-聚合酶链反应(RT-PCR)检测肾皮质ET-1、NOS的mRNA表达。
结果MS大鼠SBP、DBP、MAP均有不同程度增高,血浆和肾组织NO、NOS、和肾脏NOS基因表达水平显著下降(P<0.05),同时血浆和肾组织中ET-1和肾脏ET-1mRNA表达水平显著上升(P<0.05),经中药HQSW干预治疗后,HQSW可以改善MS大鼠的高血压状况,使实验大鼠SBP、DBP、MAP均有不同程度减低；血浆和肾皮质匀浆中NO、NOS和肾组织NOSmRNA表达上调(P<0.05),相反,血浆和肾皮质ET-1浓度和肾组织ET-1mRNA表达下调(P<0.05)。
结论MS大鼠发生NO/ET-1平衡的紊乱,从而导致血压增高。HQSW可以改善MS大鼠的高血压状况,使实验大鼠SBP、DBP、MAP均有不同程度减低,它依赖于对大鼠的ET-1/NO的动态平衡具有调节作用,这可能是其发挥防治早期MS肾损伤的主要机制之一。
第四部分复方黄芪首乌合剂MS大鼠足细胞的保护及Podocin和Nephrin表达的调控作用
目的观察HQSW对实验性代谢综合征大鼠早期足细胞损伤的保护作用并探索其作用机制。
方法用高糖高脂饲料喂养建立实验性代谢综合征大鼠模型,以HQSW(低剂量组、中剂量组和高剂量组)灌胃治疗。8周后,比色法测定实验大鼠24小时尿蛋白(24hPro)；免疫比浊法测定尿微量白蛋白(MA)和尿微量白蛋白与尿肌酐比值(albumin to creatinine ratio, ACR);免疫组织化学法检测肾组织Nephrin和Podocin的蛋白表达；逆转录-聚合酶链反应(RT-PCR)检测肾皮质Nephrin和Podocin mRNA的表达；电镜下观察肾脏基底膜(GBM)和足细胞的变化。
结果MS大鼠24hPro.MA和ACR较正常对照组明显增加(P<0.01),肾小球基底膜显著增厚,足细胞与对照组大鼠比较明显减少,足突排列紊乱,足突融合；肾小球Podocin和Nephrin的蛋白和基因表达明显低于正常对照组(P<0.05)。经HQSW干预治疗8周后,24hPro、MA、MA/UCr明显减少(P<0.05)；肾小球基底膜病变减轻,细胞外基质增多减轻,足细胞数目增多,足细胞融合减轻；肾脏皮质Podocin和Nephrin的蛋白与基因表达分别较模型组最显著增加(P<0.05)。
结论MS大鼠肾脏足细胞发生明显的病理损害,同时伴随着Nephrin和Podocin的表达异常。HQSW能防治MS大鼠的足细胞损害,修复肾组织Nephrin和Podocin的蛋白和基因的异常表达,这可能是其发挥防治MS早期肾损伤的主要机制之一。
Part I The effects of the mixture of Radix Astragalus and Polygonum mulflorum on glucose and lipid metabolism and renal function in rats with MS
Objective To investigate the effect of mixture Radix Astragalus and Polygonum mulflorum on glucose and lipid metabolism and renal function in rats with metabolic syndrome.
Methods To establish experimental metabolic syndrome rat model through a high-sugar and high-fat diet, Animals were administered with the mixture of Radix Astragalus and Polygonum mulflorum (low dose group, middle dose group and high dose group). At the8th week, the serum were used to detect TG、TCH、Scr、Bun、 UA and Glu with biochemical analyzer, the pathologic change of kidney was observed by light microscope
Results At the end of8th week,In MS model group, there is a significant increase in body weight, serum glucose, cholesterol, triglycerides and low density lipoprotein, and accompany with the significant decrease of high-density lipoprotein and insulin sensitivity index, those satisfied the diagnostic indications of MS. The body weight of rats treated with the mixture of Radix Astragalus and Polygonum mulflorum was decreased significantly compared with model group, and there is similar results in rats treated with Metformin(P＜0.05); Radix Astragalus and Polygonum mulflorum mixture can significantly reduce serum levels of TCH, TG and LDL (P＜0.05), at the same time the serum level of HDL-C increased significantly (P＜0.05); there is significant change on Scr、BUN and UA in experimental rat treated with the mixture of Radix Astragalus and Polygonum mulflorum (P<0.05), the pathologic change was improved after treatment.
Conclusion:The model of metabolic syndrome in rats can be successfully replicated through High-sugar and high-fat diet. The mixture of Radix Astragalus and Polygonum mulflorum can improve insulin sensitivity, and regulate lipid metabolism, reduce blood glucose, and protect renal function. Above all it can improve the pathologic change in MS rats.
Part II The effects of the mixture of Radix Astragalus and Polygonum mulflorum on renal secretion of AngⅡ and FFA and oxidative stress in rats with MS
Objective To investigate the effect of the mixture of Radix Astragalus and Polygonum mulflorum on renal secretion of angiotensin Ⅱ (AngⅡ) and free fatty acids (FFA) and oxidative stress in rats with metabolic syndrome.
Methods To establish experimental metabolic syndrome rat model through a high-sugar and high-fat diet, Animals were administered with the mixture of Radix Astragalus and Polygonum mulflorum (low dose group, middle dose group and high dose group). At the end of8th week, the level of Super Oxygen Dehydrogenises (SOD) and Methane Dicarboxylic Aldehyde (MDA) in the serum and renal cortex were determined by colorimetric method, the level of AngⅡ and FFA in the serum and renal cortex were detected by radioimmunoassay.
Results The enzyme activity of SOD in the serum and renal cortex was decreased significantly in rats with MS (P＜0.01), on the contrary, the level of MDA、AngⅡ and FFA in the serum and renal cortex were increased significantly (P＜0.01). After treated with the mixture of Radix Astragalus and Polygonum mulflorum, The enzyme activity of SOD in the serum and renal cortex was increased significantly (P＜0.05), At the same time, The level of MDA、AngⅡ and FFA in the serum and renal cortex were decreased significantly (P＜0.05)
Conclusion The abnormal secretion of FFA and AngⅡ were occurred in rats with MS, at the same time, the oxidative stress response was being aggravated. After treated with the mixture of Radix Astragalus and Polygonum mulflorum, the above abnormal change was improved, that might be its prevention and treatment mechanism of early renal injury in rats with MS.
Part Ⅲ The regulation of mixture of Radix Astragalus and Polygonum mulflorum on balance of NO and ET-1in rats with MS
Objective To investigate the effect on blood pressure and regulation of the mixture of Radix Astragalus and Polygonum mulflorum on the balance of Nitric Oxide (NO) and endothelin-1(ET-1) in rats with metabolic syndrome.
Methods To establish experimental metabolic syndrome rat model through a high-sugar and high-fat diet, Animals were administered with Radix Astragalus compound mixture (low dose group, middle dose group and high dose group) for8weeks. Then, the level of SBP、DBP、MAP were detected with multifunction physiological monitor, The level of NO and the enzyme activity of NOS in the serum and renal cortex were determined by colorimetric method, he level of ET-1in the serum and renal cortex were detected by radioimmunoassay, The mRNA expressions of Renal ET-1and NOS were detected through reverse transcription polymerase chain reaction(RT-PCR).
Results The level of SBP、DBP、MAP in MS rats was increased significantly compared with normal rats (P＜0.05), The enzyme activity of NOS and the level of NO in the serum and renal cortex and the renal gene expression of NOS was decreased significantly in rats with MS (P＜0.05), On the contrary, the level of ET-1in the serum and renal cortex and the renal gene expression of ET-1was increased significantly (P＜0.05). After treated with the mixture of Radix Astragalus and Polygonum mulflorum, The level of SBP、DBP、MAP in MS rats was decreased significantly compared with normal rats (P＜0.05),The enzyme activity of NOS and the level of NO in the serum and renal cortex and the renal gene expression of NOS was increased significantly (P＜0.05)(P＜0.05), At the same time, the level of ET-1in the serum and renal cortex and the renal gene expression of ET-1was decreased significantly (P＜0.05)
Conclusion The balance of NO/ET-1was destroyed in rats with MS, and then the abnormal level of SBP、DBP、MAP were detected, The mixture of Radix Astragalus and Polygonum mulflorum can lower the blood pression in MS rats through to regulate the balance of NO/ET-1in MS rats, this might play key role in protection the early renal injury of MS rats.
Part IV The effects of the mixture of Radix Astragalus and Polygonum mulflorum on renal expression of Podocin and Nephrin in rats with MS
Objective To investigate the effects and mechanism of the mixture of Radix Astragalus and Polygonum mulflorum on early injury of renal podocyte in rats with metabolic syndrome.
Methods To establish experimental metabolic syndrome rat model through a high-sugar and high-fat diet, Animals were administered with Radix Astragalus compound mixture (low dose group, middle dose group and high dose group) for8weeks. Then, the level of24hPro was detected by colorimetric method; the level of MA and ACR were detected by lmmunoturbidimetry; the protein expression of Nephrin and Podocin in renal cortex was determined by immunohistochemistry; the gene expression of Nephrin and Podocin in renal cortex was determined by RT-PCR; The pathological change of Glomerular Basement Membrane(GBM) and podocyte was observed by electron microscope.
Results The level of24hPro、MA and ACR were increased significantly in rats with MS compared with normal rats(P＜0.05); Under electronic microscopic examination, fusion of foot process, disruption of GBM and enlargement of infiltration pores, thickness of GBM were observed; the protein and gene expression of Podocin and Nephrin in renal cortex was decreased significantly compared with normal rats(P＜0.05). After8weeks treatment with the mixture of Radix Astragalus and Polygonum mulflorum, The level of24hPro、MA and ACR were decreased significantly in rats with MS compared with normal rats(P＜0.05); the pathologic change of GBM and podocte was reduced significantly compared with MS rats; the protein and gene expression of Podocin and Nephrin in renal cortex was increased significantly compared with normal rats(P＜0.05).
Conclusion The pathological change of podocyte was observed in rats with MS, accompanied by the abnormal protein and gene expression of Nephrin and Podocin in renal cortex. After the treatment of the mixture of Radix Astragalus and Polygonum mulflorum, the abnormal condition was improved, and it might play a key role in protection the early renal injury of MS rats.
引文
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