竹笋降压降脂有效成分及其活性研究
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摘要
竹笋(Bamboo shoots)是我国的大宗林副产品,水煮笋罐头是竹笋加工的主要产品形态。传统的水煮笋罐头是指以新鲜竹笋为原料,经去壳、漂洗、煮制等初级加工方式处理后,再经罐装、密封、杀菌等工艺制成的竹笋罐头。大罐的水煮笋罐头除直接进入流通领域以外,也常在工厂中作为小包装产品(尤其是方便调味笋制品)的原料。上述加工过程伴随着大量废弃物的产生,给产地带来很大的环保压力。本论文以水煮笋加工过程中的大宗废弃物(包括笋煮液、烫漂液、罐内液、下脚料压榨汁等,统称笋液)的开发利用为切入点,瞄准笋液中丰富的氨基酸、肽、多糖、黄酮和酚酸等次生代谢产物,以期挖掘出有益于人类健康的天然功能性素材。本论文以竹笋ACE抑制剂为重点研究对象,对其分离制备工艺、特征性成分的化学结构、体外ACE抑制活性以及体内降压、降脂功能进行了系统研究。现将主要研究成果归纳如下：
1、经过预处理的浓缩笋液(AEBS)富含氨基酸(包括肽),含量为21.32±1.16g/100g干基；其次是多糖,含量为13.17+0.98g/100g干基；此外,还含有一定量的黄酮和酚酸类化合物,总黄酮含量为1.41±0.12g/100g干基,酚酸含量为4.78+0.24g/100g干基。以AEBS为出发物料,以体外血管紧张素转化酶(ACE)抑制活性和抗氧化活性(DPPH·、 ABTS·+、FRAP)为评价和示踪指标,采用膜分离和大孔树脂吸附-解吸等技术进行精制,分别得到了具有较佳体外ACE抑制活性和抗氧化性能的竹笋ACE抑制剂(BSP)和竹笋粗多糖(多糖含量68.73±3.24g/100g)二个有效活性部位。
2、通过对从AEBS中得到的竹笋粗多糖(膜截留分子量大于10KDa)进行水解,结果表明,水解后总黄酮含量没有显著变化,但总酚含量大幅度增加。其中,酸水解样品总酚含量显著高于碱水解样品,尤其是对香豆酸和阿魏酸含量显著增加。这可能是由于在酸水解过程中,竹笋粗多糖中的多糖酚酸酯解构释放出游离酚酸所致。与粗多糖及其碱水解样品相比,酸水解样品表现出更强的抗氧化性能和体外ACE抑制活性。同时,结果还表明不同试样中总酚含量与其抗氧化活性和体外ACE抑制活性呈显著正相关。
3、采用乙酸乙酯和正丁醇对竹笋ACE抑制剂(BSP)的水溶液进行分级萃取,乙酸乙酯相(BSME)表现出最强的抗氧化活性,水分级相(BSML)表现出最强的体外ACE抑制活性(即剩余相)。对乙酸乙酯相(BSME)、正丁醇相(BSMB)和水分级相(BSML)进行抗氧化和体外ACE抑制活性评价,结果表明抗氧化活性与黄酮含量呈正相关,而氨基酸含量与体外ACE抑制活性呈正相关。以水分级相(BSML)为出发材料,以体外ACE抑制活性为指征,采用Sephadex G-15凝胶对BSML进一步分离纯化,得到活性最强的部位P2。以半制备反相高效液相色谱对P2组分进行分离制备,得到高活性组分P2-2,采用UPLC-ESI-MS进行结构解析,确定活性二肽为天冬氨酸-酪氨酸(Asp-Tyr)。委托相关机构,对上述特征性竹笋肽进行化学合成,获得了g级Asp-Tyr二肽试样。
4、竹笋ACE抑制剂(BSP)中富含氨基酸、肽和多酚类化合物,质量指标为：氨基酸总量42.78±0.57g/100g,总黄酮含量5.07±0.47g RE/100g,酚酸含量为10.91±0.98gGAE/100g。对BSP改善高脂血大鼠心血管功能和氧化应激试验研究表明,高剂量BSP (350mg/day kg BW)能显著降低高脂血症大鼠血清总胆固醇(Total cholesterol, TC)、甘油三酯(Triacylglycerol, TG)、低密度脂蛋白胆固醇(Low-density lipoprotein cholesterol, LDL-c)含量和动脉粥样硬化指数(Atherosclerosis index, AI),抑制肝脏中脂肪酸合成酶(FAS)活性；中剂量BSP (200mg/day kg BW)能有效降低大鼠肝脏TC、TG水平；低剂量BSP (100mg/day kg BW)能有效降低大鼠肝脏TC水平。各剂量BSP组均可显著改善高脂血大鼠的氧化应激状态,提高机体内源性超氧化物歧化酶(Superoxide dismutase, SOD)和谷胱甘肽过氧化物(Glutathione peroxidase, GSH-Px)活力,降低肝脏丙二醛(Malonaldehyde, MDA)含量。
5、本部分实验以自发性高血压大鼠(Spontaneously hypertensive rats, SHRs)为试验模型评价BSP的降压作用。一次性降压实验表明,给药4h-6h后,100mg/day kg BW剂量BSP降压效果与10mg/day kg BW剂量的阳性对照药物卡普托利(Captopril)接近；50mg/day kg BW BSP降压效果与10mg/day kg BW合成二肽Asp-Tyr相当。在上述试验的基础上,对SHR大鼠进行连续四周的降压试验,设以下六个试验组：SHR对照组、阳性对照组(Captopril,10mg/day kg BW), BSP二个剂量组(50mg/day kg BW和100mg/day kg BW)合成肽组(Asp-Tyr,10mg/day kg BW)以及BSP和竹茹三萜(EZR2002)复配组(BSP50mg/day kg BW+EZR200250mg/day kg BW)。试验结果表明：(1)连续给药不同剂量的BSP7d后,大鼠血压开始下降,21d实验组血压与空白组对比差异明显；BSP与EZR2002复配协同增效降压效果最为显著,显示出良好的降压效果。(2)不同剂量BSP均能有效抑制SHR大鼠肺部组织中的ACE活性,并呈现剂量依赖关系(p<0.05)。BSP与EZR2002复配协同增效,显著抑制肺部ACE活性(p<0.01)。(3)合成肽(10mg/day kg BW)显著提高血清GSH-Px活性(p<0.05)；不同受试剂量的BSP均可显著提高显著升高血清SOD、GSH-Px酶活力和肝脏总抗氧化能力,升高血清一氧化氮(Nitric oxide, NO)水平,增加肾脏一氧化氮合酶(Nitric oxide synthase, NOS)活性,并呈现剂量效应；BSP与EZR2002复配可显著增加血清SODD(p<0.01)、GSH-Px(p<0.01)活力,肾脏NOS活性(p<0.01),降低血清和肝脏MDA水平(p<0.01)。
本研究结果表明,源自罐头笋加工废液的功能性食品素材——竹笋ACE抑制剂(BSP)富含氨基酸、寡肽、黄酮和酚酸类化合物等有效成分,是竹笋的最佳有效活性部位。体内外试验研究均表明,BSP具有高效ACE抑制活性和显著的降压、降脂、抗氧化功能,能显著改善高脂血大鼠脂质代谢和氧化应激状态、降低SHR大鼠的血压、改善SHR大鼠的氧化应激状态。鉴于我国丰富的竹笋资源及其巨大的笋加工副产物总量,从竹笋中挖掘天然来源的ACE抑制剂具有特殊的理论意义和现实意义,既能为心血管疾病的膳食防治找到新的手段和途径,同时又能推动竹产业的资源转化和技术进步。
Bamboo shoot, the young stems of bamboo, is a kind of common vegetables for centuries in China. Phyllostachys pubescens, which is widely distributed in China, is the main bamboo of producing bamboo shoot. Canned bamboo shoot is the main commercial product besides fresh edible. Traditional canned bamboo shoots are usually made from fresh bamboo shoot with shelled, rinsed, cooked, canned and sterilized. Large pot of canned bamboo shoot often made into small package besides directly into circulation. This process also is prone to cause serious environmental pollution, with excess boiled liquids of bamboo shoots.
In this work, our attention has been focused on the functional ingredients in boiled water of bamboo shoots, filled liquid, squeezed juice (collectively referred abstract of bamboo shoot, called AEBS in our study). We focused on AEBS, which is rich in active peptides, amino acid, polysaccharide, flavonoids and phenolics, and aimed to dig out certain natural ingredients beneficial to human health. Bamboo shoot peptides were undergone systematic chemical, technology and functional studies, in order to utilize bamboo shoots in more reasonable way. The functional roles in reducing blood lipid and lowering blood pressure of bamboo shoot peptides were investigated, with main results summarized as follows.
1) The pretreatment of AEBS was collected which is rich in amino acid (21.32±1.16g/100g, dry basis), polysaccharide (13.17±0.98g/100g, dry basis), total flavonoids (1.41±0.12g/100g, dry basis) and phenolic acids (4.78±0.24g/100g, dry basis). We selected active section, which was rich bamboo shoot peptides, using angiotensin converting enzyme (ACE, EC3.4.15.1) inhibitoiy activity and anti-oxidant system (DPPH·, ABTS·+, FRAP) as indicators. Ultra-filtration of membrane cut-off technology and macroporous resin DA201-C enrichment technology were applied, in order to enrichment active ingredients. During this process bamboo shoot ACE inhibitory preparation (BSP) and crude polysaccharide were obtained. Crude polysaccharide was obtained by ethanol precipitation from molecular weight cut-off membrane more than10KDa, which was the combination compound of phenolic components and polysaccharide polymers.
2) Crude polysaccharide polymer was hydrolysated under acid and alkali condition, respectively. The results come out that total flavonoids contents didn't exhibit significant differences before and after the hydrolysis. However, total phenolics contents were higher in the sample after acid hydrolysis, particularly for the content of p-coumaric acid and ferulic acid, which may cause by the decomposition of phenolic components and polysaccharide polymer during the acid hydrolysis. Furthermore, total phenolics contents were significantly positively correlated with the antioxidant and ACE inhibitory activities, which was the reason that the sample of acid hydrolysis exhibited the better antioxidant and higher ACE inhibitory activities compared with crude sample and hydrolysated sample under alkali condition.
3) Fractional extraction of ethyl acetate and n-butanol was applied to separate bamboo shoot ACE inhibit peptides. The result come out that the left water fraction exhibited the highest ACE inhibitoiy activity. All fractions, including ethyl acetate fraction (BSME), n-butanol fraction (BSMB) and the water fraction (BSML), were evaluated by antioxidant capacities. Results indicated that antioxidant capacities were related to total flavonoids contents, while ACE inhibitory activity was related to the content of peptides. P2, which possessed higher ACE inhibitory activity, was obtained though Sephadex G-15gel and purified further by semi-preparative reversed-phase high performance liquid chromatography. The active fraction P2-2was obtained, key component of which was dipeptide with effective sequence detected to be Asp-Tyr-by UPLC-ESI-MS.
4) BSP is rich in high contents of amino aicds (42.78±5.75g/100g), total flavonoids (5.07±0.47g RE/100g), phenolic acids (10.91±0.98g GAE/100g). Experiment was applied to investigate the effect of BSP on improving the cardiovascular status and oxidative stress status in hyperlipidemic rats. Results showed that350mg/day kg BW BSP administration could significantly reduce content of total cholesterol (TC), triacylglycerol (TG), low-density lipoprotein cholesterol (LDL-c) in the serum of hyperlipidemia rats, the atherosclerosis index (AI) and fatty acid synthase (FAS) in Liver.200mg/day kg BW BSP administration could effectively reduce the content of TC, TG and LDL-c in the liver of hyperlipidemia rats, while100mg/day kg BW BSP administration could only reduce content of TC in the liver of hyperlipidemia rats. All administration groups could significantly increase the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and reduce content of malonaldehyde (MDA), which meaned that the oxidative stress status had been alleviated.
5) Spontaneously hypertensive rats (SHR) was applied as test model for evaluation the biology effect of BSP. In the (0～8h) experiment of anti-hypertension in SHR,100mg/day kg BW BSP exhibited equivalent effect to10mg/day kg BW Captoril after4h～6h administration.50mg/day kg BW BSP exhibited equivalent effect to10mg/day kg BW synthetic peptides.
Based on above test,4weeks experiment of BSP regulation blood pressure was carried out: SHR control group (distilled water), positive control group (Captopril,10mg/day kg BW), BSP (50mg/day kg BW), BSP (100mg/day kg BW), Combined group of BSP and bamboo shavings (EZR2002)(BSP50mg/day kg BW+EZR200250mg/day kg BW).
The blood pressure of SHR decreased after7d BSP (50mg/day kg BW,100mg/day kg BW) administration, compared with control group. BSP (50mg/day kg BW,100mg/day kg BW) could inhibit ACE activity in lung and in serum with dose effective. The combination of BSP50mg/day kg BW and EZR200250mg/day kg BW showed excellent effect on anti-hypertensive and inhibited ACE activity in lung of SHR (p<0.01) and in the serum (p<0.01).
The oxidative stress status was evaluated. Synthetic peptide could significantly increase the serum GSH-Px activity (p<0.05) of SHRs. BSP (50mg/day kg BW,100mg/day kg BW) could increase the activities of serum SOD, GSH-Px, NO, NOS in kidney and total anti-oxidant capaticy in liver with dose effective. The combination of BSP50mg/day kg BW and EZR200250mg/day kg BW showed excellent effect on oxidative stress status:increase the activities of serum SOD (p<0.01), GSH-Px (p<0.01), NO(p<0.01), NOS in kidney (p<0.01), MDA contents both in the serum (p<0.01)and liver (p<0.01).
In conclusion, BSP and EZR2002derived from bamboo secondary metabolites exhibited prominent capacity to improve oxidative stress status in hyperlipidemic rats and SHRs. The antihyperlipidemic effect was attributed to phenolic compounds via FAS inhibition. Antihypertension experiment effect contributes to ACE inhibition and improving NO level. Howerver, the regulation mechanism and related signal pathways need further research and investigation. The active ingredients were enrichment by combination of ultra-filtration of cut-off membrane technology and macroporous resin DA201-C technology, which could provide reference for further utilizatin of by-product from canned bamboo shoot industry.
1、经过预处理的浓缩笋液(AEBS)富含氨基酸(包括肽),含量为21.32±1.16g/100g干基；其次是多糖,含量为13.17+0.98g/100g干基；此外,还含有一定量的黄酮和酚酸类化合物,总黄酮含量为1.41±0.12g/100g干基,酚酸含量为4.78+0.24g/100g干基。以AEBS为出发物料,以体外血管紧张素转化酶(ACE)抑制活性和抗氧化活性(DPPH·、 ABTS·+、FRAP)为评价和示踪指标,采用膜分离和大孔树脂吸附-解吸等技术进行精制,分别得到了具有较佳体外ACE抑制活性和抗氧化性能的竹笋ACE抑制剂(BSP)和竹笋粗多糖(多糖含量68.73±3.24g/100g)二个有效活性部位。
2、通过对从AEBS中得到的竹笋粗多糖(膜截留分子量大于10KDa)进行水解,结果表明,水解后总黄酮含量没有显著变化,但总酚含量大幅度增加。其中,酸水解样品总酚含量显著高于碱水解样品,尤其是对香豆酸和阿魏酸含量显著增加。这可能是由于在酸水解过程中,竹笋粗多糖中的多糖酚酸酯解构释放出游离酚酸所致。与粗多糖及其碱水解样品相比,酸水解样品表现出更强的抗氧化性能和体外ACE抑制活性。同时,结果还表明不同试样中总酚含量与其抗氧化活性和体外ACE抑制活性呈显著正相关。
3、采用乙酸乙酯和正丁醇对竹笋ACE抑制剂(BSP)的水溶液进行分级萃取,乙酸乙酯相(BSME)表现出最强的抗氧化活性,水分级相(BSML)表现出最强的体外ACE抑制活性(即剩余相)。对乙酸乙酯相(BSME)、正丁醇相(BSMB)和水分级相(BSML)进行抗氧化和体外ACE抑制活性评价,结果表明抗氧化活性与黄酮含量呈正相关,而氨基酸含量与体外ACE抑制活性呈正相关。以水分级相(BSML)为出发材料,以体外ACE抑制活性为指征,采用Sephadex G-15凝胶对BSML进一步分离纯化,得到活性最强的部位P2。以半制备反相高效液相色谱对P2组分进行分离制备,得到高活性组分P2-2,采用UPLC-ESI-MS进行结构解析,确定活性二肽为天冬氨酸-酪氨酸(Asp-Tyr)。委托相关机构,对上述特征性竹笋肽进行化学合成,获得了g级Asp-Tyr二肽试样。
4、竹笋ACE抑制剂(BSP)中富含氨基酸、肽和多酚类化合物,质量指标为：氨基酸总量42.78±0.57g/100g,总黄酮含量5.07±0.47g RE/100g,酚酸含量为10.91±0.98gGAE/100g。对BSP改善高脂血大鼠心血管功能和氧化应激试验研究表明,高剂量BSP (350mg/day kg BW)能显著降低高脂血症大鼠血清总胆固醇(Total cholesterol, TC)、甘油三酯(Triacylglycerol, TG)、低密度脂蛋白胆固醇(Low-density lipoprotein cholesterol, LDL-c)含量和动脉粥样硬化指数(Atherosclerosis index, AI),抑制肝脏中脂肪酸合成酶(FAS)活性；中剂量BSP (200mg/day kg BW)能有效降低大鼠肝脏TC、TG水平；低剂量BSP (100mg/day kg BW)能有效降低大鼠肝脏TC水平。各剂量BSP组均可显著改善高脂血大鼠的氧化应激状态,提高机体内源性超氧化物歧化酶(Superoxide dismutase, SOD)和谷胱甘肽过氧化物(Glutathione peroxidase, GSH-Px)活力,降低肝脏丙二醛(Malonaldehyde, MDA)含量。
5、本部分实验以自发性高血压大鼠(Spontaneously hypertensive rats, SHRs)为试验模型评价BSP的降压作用。一次性降压实验表明,给药4h-6h后,100mg/day kg BW剂量BSP降压效果与10mg/day kg BW剂量的阳性对照药物卡普托利(Captopril)接近；50mg/day kg BW BSP降压效果与10mg/day kg BW合成二肽Asp-Tyr相当。在上述试验的基础上,对SHR大鼠进行连续四周的降压试验,设以下六个试验组：SHR对照组、阳性对照组(Captopril,10mg/day kg BW), BSP二个剂量组(50mg/day kg BW和100mg/day kg BW)合成肽组(Asp-Tyr,10mg/day kg BW)以及BSP和竹茹三萜(EZR2002)复配组(BSP50mg/day kg BW+EZR200250mg/day kg BW)。试验结果表明：(1)连续给药不同剂量的BSP7d后,大鼠血压开始下降,21d实验组血压与空白组对比差异明显；BSP与EZR2002复配协同增效降压效果最为显著,显示出良好的降压效果。(2)不同剂量BSP均能有效抑制SHR大鼠肺部组织中的ACE活性,并呈现剂量依赖关系(p<0.05)。BSP与EZR2002复配协同增效,显著抑制肺部ACE活性(p<0.01)。(3)合成肽(10mg/day kg BW)显著提高血清GSH-Px活性(p<0.05)；不同受试剂量的BSP均可显著提高显著升高血清SOD、GSH-Px酶活力和肝脏总抗氧化能力,升高血清一氧化氮(Nitric oxide, NO)水平,增加肾脏一氧化氮合酶(Nitric oxide synthase, NOS)活性,并呈现剂量效应；BSP与EZR2002复配可显著增加血清SODD(p<0.01)、GSH-Px(p<0.01)活力,肾脏NOS活性(p<0.01),降低血清和肝脏MDA水平(p<0.01)。
本研究结果表明,源自罐头笋加工废液的功能性食品素材——竹笋ACE抑制剂(BSP)富含氨基酸、寡肽、黄酮和酚酸类化合物等有效成分,是竹笋的最佳有效活性部位。体内外试验研究均表明,BSP具有高效ACE抑制活性和显著的降压、降脂、抗氧化功能,能显著改善高脂血大鼠脂质代谢和氧化应激状态、降低SHR大鼠的血压、改善SHR大鼠的氧化应激状态。鉴于我国丰富的竹笋资源及其巨大的笋加工副产物总量,从竹笋中挖掘天然来源的ACE抑制剂具有特殊的理论意义和现实意义,既能为心血管疾病的膳食防治找到新的手段和途径,同时又能推动竹产业的资源转化和技术进步。
Bamboo shoot, the young stems of bamboo, is a kind of common vegetables for centuries in China. Phyllostachys pubescens, which is widely distributed in China, is the main bamboo of producing bamboo shoot. Canned bamboo shoot is the main commercial product besides fresh edible. Traditional canned bamboo shoots are usually made from fresh bamboo shoot with shelled, rinsed, cooked, canned and sterilized. Large pot of canned bamboo shoot often made into small package besides directly into circulation. This process also is prone to cause serious environmental pollution, with excess boiled liquids of bamboo shoots.
In this work, our attention has been focused on the functional ingredients in boiled water of bamboo shoots, filled liquid, squeezed juice (collectively referred abstract of bamboo shoot, called AEBS in our study). We focused on AEBS, which is rich in active peptides, amino acid, polysaccharide, flavonoids and phenolics, and aimed to dig out certain natural ingredients beneficial to human health. Bamboo shoot peptides were undergone systematic chemical, technology and functional studies, in order to utilize bamboo shoots in more reasonable way. The functional roles in reducing blood lipid and lowering blood pressure of bamboo shoot peptides were investigated, with main results summarized as follows.
1) The pretreatment of AEBS was collected which is rich in amino acid (21.32±1.16g/100g, dry basis), polysaccharide (13.17±0.98g/100g, dry basis), total flavonoids (1.41±0.12g/100g, dry basis) and phenolic acids (4.78±0.24g/100g, dry basis). We selected active section, which was rich bamboo shoot peptides, using angiotensin converting enzyme (ACE, EC3.4.15.1) inhibitoiy activity and anti-oxidant system (DPPH·, ABTS·+, FRAP) as indicators. Ultra-filtration of membrane cut-off technology and macroporous resin DA201-C enrichment technology were applied, in order to enrichment active ingredients. During this process bamboo shoot ACE inhibitory preparation (BSP) and crude polysaccharide were obtained. Crude polysaccharide was obtained by ethanol precipitation from molecular weight cut-off membrane more than10KDa, which was the combination compound of phenolic components and polysaccharide polymers.
2) Crude polysaccharide polymer was hydrolysated under acid and alkali condition, respectively. The results come out that total flavonoids contents didn't exhibit significant differences before and after the hydrolysis. However, total phenolics contents were higher in the sample after acid hydrolysis, particularly for the content of p-coumaric acid and ferulic acid, which may cause by the decomposition of phenolic components and polysaccharide polymer during the acid hydrolysis. Furthermore, total phenolics contents were significantly positively correlated with the antioxidant and ACE inhibitory activities, which was the reason that the sample of acid hydrolysis exhibited the better antioxidant and higher ACE inhibitory activities compared with crude sample and hydrolysated sample under alkali condition.
3) Fractional extraction of ethyl acetate and n-butanol was applied to separate bamboo shoot ACE inhibit peptides. The result come out that the left water fraction exhibited the highest ACE inhibitoiy activity. All fractions, including ethyl acetate fraction (BSME), n-butanol fraction (BSMB) and the water fraction (BSML), were evaluated by antioxidant capacities. Results indicated that antioxidant capacities were related to total flavonoids contents, while ACE inhibitory activity was related to the content of peptides. P2, which possessed higher ACE inhibitory activity, was obtained though Sephadex G-15gel and purified further by semi-preparative reversed-phase high performance liquid chromatography. The active fraction P2-2was obtained, key component of which was dipeptide with effective sequence detected to be Asp-Tyr-by UPLC-ESI-MS.
4) BSP is rich in high contents of amino aicds (42.78±5.75g/100g), total flavonoids (5.07±0.47g RE/100g), phenolic acids (10.91±0.98g GAE/100g). Experiment was applied to investigate the effect of BSP on improving the cardiovascular status and oxidative stress status in hyperlipidemic rats. Results showed that350mg/day kg BW BSP administration could significantly reduce content of total cholesterol (TC), triacylglycerol (TG), low-density lipoprotein cholesterol (LDL-c) in the serum of hyperlipidemia rats, the atherosclerosis index (AI) and fatty acid synthase (FAS) in Liver.200mg/day kg BW BSP administration could effectively reduce the content of TC, TG and LDL-c in the liver of hyperlipidemia rats, while100mg/day kg BW BSP administration could only reduce content of TC in the liver of hyperlipidemia rats. All administration groups could significantly increase the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and reduce content of malonaldehyde (MDA), which meaned that the oxidative stress status had been alleviated.
5) Spontaneously hypertensive rats (SHR) was applied as test model for evaluation the biology effect of BSP. In the (0～8h) experiment of anti-hypertension in SHR,100mg/day kg BW BSP exhibited equivalent effect to10mg/day kg BW Captoril after4h～6h administration.50mg/day kg BW BSP exhibited equivalent effect to10mg/day kg BW synthetic peptides.
Based on above test,4weeks experiment of BSP regulation blood pressure was carried out: SHR control group (distilled water), positive control group (Captopril,10mg/day kg BW), BSP (50mg/day kg BW), BSP (100mg/day kg BW), Combined group of BSP and bamboo shavings (EZR2002)(BSP50mg/day kg BW+EZR200250mg/day kg BW).
The blood pressure of SHR decreased after7d BSP (50mg/day kg BW,100mg/day kg BW) administration, compared with control group. BSP (50mg/day kg BW,100mg/day kg BW) could inhibit ACE activity in lung and in serum with dose effective. The combination of BSP50mg/day kg BW and EZR200250mg/day kg BW showed excellent effect on anti-hypertensive and inhibited ACE activity in lung of SHR (p<0.01) and in the serum (p<0.01).
The oxidative stress status was evaluated. Synthetic peptide could significantly increase the serum GSH-Px activity (p<0.05) of SHRs. BSP (50mg/day kg BW,100mg/day kg BW) could increase the activities of serum SOD, GSH-Px, NO, NOS in kidney and total anti-oxidant capaticy in liver with dose effective. The combination of BSP50mg/day kg BW and EZR200250mg/day kg BW showed excellent effect on oxidative stress status:increase the activities of serum SOD (p<0.01), GSH-Px (p<0.01), NO(p<0.01), NOS in kidney (p<0.01), MDA contents both in the serum (p<0.01)and liver (p<0.01).
In conclusion, BSP and EZR2002derived from bamboo secondary metabolites exhibited prominent capacity to improve oxidative stress status in hyperlipidemic rats and SHRs. The antihyperlipidemic effect was attributed to phenolic compounds via FAS inhibition. Antihypertension experiment effect contributes to ACE inhibition and improving NO level. Howerver, the regulation mechanism and related signal pathways need further research and investigation. The active ingredients were enrichment by combination of ultra-filtration of cut-off membrane technology and macroporous resin DA201-C technology, which could provide reference for further utilizatin of by-product from canned bamboo shoot industry.
引文
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