【摘要】：連錢草[Glechoma longituba(Nakai)Kupr.]為唇形科活血丹屬草本植物,具有溶石、抗菌、消炎等作用。本文對(duì)連錢草多糖進(jìn)行研究,優(yōu)化了連錢草多糖的脫蛋白脫色素工藝,分離純化出分子量均一分布的多糖并對(duì)其理化性質(zhì)、結(jié)構(gòu)組成及生物活性進(jìn)行了研究,為連錢草多糖的功能性開(kāi)發(fā)奠定了基礎(chǔ)。1.通過(guò)水提醇沉法獲取連錢草粗多糖(GP),以多糖保留率、蛋白質(zhì)清除率和脫色率為指標(biāo),在單因素試驗(yàn)的基礎(chǔ)上采用正交試驗(yàn)分別考察了活性炭、大孔樹(shù)脂D301G對(duì)GP溶液的脫蛋白脫色素效果,確定最優(yōu)脫蛋白脫色素工藝。結(jié)果表明,活性炭對(duì)GP的最佳脫蛋白脫色素條件為:活性炭添加量1.0%(g/mL),振蕩時(shí)間1.5 h,多糖濃度6 mg/mL,樣液pH 3.0,此條件下多糖保留率、蛋白質(zhì)清除率、脫色率分別為(95.32±1.66)%、(94.59±0.34)%、(86.49±0.81)%;D301G對(duì)GP溶液的最佳脫蛋白脫色素條件為:大孔樹(shù)脂添加量15%(g/mL),振蕩時(shí)間0.5 h,多糖濃度2 mg/mL,樣液pH 7.0,此條件下多糖保留率、蛋白質(zhì)清除率、脫色率分別為(93.41±0.87)%、(95.82±0.19)%、(96.97±0.05)%;綜合比較,D301G對(duì)GP的脫蛋白脫色素效果優(yōu)于活性炭。脫蛋白脫色素后的粗多糖GPC依次經(jīng)DEAE Sephadex A-25離子交換層析和Sephacryl S-200凝膠過(guò)濾層析,得到三個(gè)組分:GRa-1、GRb-1、GRb-2。2.考察了GRa-1、GRb-1、GRb-2的理化性質(zhì)和純度,并分析了GRa-1的單糖組成及結(jié)構(gòu)特點(diǎn)。結(jié)果表明,GRa-1、GRb-1為中性多糖,GRb-2為堿性多糖,三者均不含蛋白質(zhì);HPLC分析顯示GRa-1為分子量均一分布的多糖,相對(duì)分子質(zhì)量為3 584 Da,而GRb-1和GRb-2則含多種多糖組分,有待進(jìn)一步純化;GRa-1的紅外圖譜顯示該多糖含有D型甘露呋喃糖;GC-MS分析GRa-1的單糖組成為阿拉伯糖、甘露糖、葡萄糖、半乳糖四種,摩爾比為0.005:0.014:0.168:0.813。3.研究連錢草多糖分離純化中各組分的體外抗氧化活性和分布。結(jié)果表明,粗多糖GP和GPC在各方面都顯示出較強(qiáng)的體外抗氧化活性;GRa-1在Fe2+螯合及O-2·、OH·、DPPH+·清除上表現(xiàn)出較強(qiáng)的活性;GRb-1對(duì)O-2·和OH·表現(xiàn)出較強(qiáng)的清除能力;GRb-2除Fe2+螯合和O-2·、OH·、DPPH+·清除力較強(qiáng)外還具有強(qiáng)的總還原力。4.通過(guò)腹腔注射葡萄糖耐量試驗(yàn)(Intraperitoneal Glucose Tolerance Test,IPGTT)、口服脂肪耐量試驗(yàn)(Oral Fat Tolerance Test,OFTT)、二甲苯致小鼠耳腫脹試驗(yàn)、老年小鼠抗氧化試驗(yàn),檢測(cè)了GRa-1的降血糖、降血脂、抗炎和體內(nèi)抗氧化活性,結(jié)果表明,GRa-1能顯著提高小鼠對(duì)葡萄糖的利用能力;加速小鼠體內(nèi)甘油三酯的代謝;顯著抑制二甲苯致小鼠耳腫脹;增強(qiáng)老年小鼠血清和肝臟中部分抗氧化酶活性,降低脂質(zhì)過(guò)氧化產(chǎn)物(MDA)含量。
[Abstract]:Glechoma longituba (Nakai) Kupr. It is a herbaceous plant of Huoxue Dan genus in Lip family. It has the functions of litholysis, antibacterial, anti-inflammatory and so on. In this paper, we studied the polysaccharides of Polygonum chinensis, optimized the deproteinization and depigmentation process, isolated and purified the polysaccharides with uniform molecular weight distribution, and studied their physicochemical properties, structural composition and biological activities. It lays a foundation for the functional development of polysaccharides. 1. The crude polysaccharide (GP), obtained by water extraction and alcohol precipitation was characterized by polysaccharide retention rate, protein clearance rate and decolorization rate. On the basis of single factor experiment, the activated carbon was investigated by orthogonal test. The effect of macroporous resin D301G on the depigmentation of GP solution was studied. The results showed that the optimum conditions for deproteinization of GP by activated carbon were as follows: the dosage of activated carbon was 1.0% (g/mL), the oscillating time was 1.5 h, and the concentration of polysaccharide was 6 mg/mL, sample solution pH 3.0. Under this condition, the polysaccharide retention rate and protein clearance rate were obtained. The decolorization rates were (95.32 鹵1.66), (94.59 鹵0.34), (86.49 鹵0.81) and (86.49 鹵0.81), respectively. The optimum conditions for decolorization of GP solution were as follows: the amount of macroporous resin was 15% (g/mL), the oscillating time was 0.5 h, and the concentration of polysaccharide was 2 mg/mL, like solution pH 7.0. The decolorization rates were (93.41 鹵0.87), (95.82 鹵0.19), (96.97 鹵0.05), respectively. The crude polysaccharide GPC was separated by DEAE Sephadex A-25 ion exchange chromatography and Sephacryl S-200 gel filtration chromatography. The physicochemical properties and purity of GRa-1,GRb-1,GRb-2 were investigated, and the monosaccharide composition and structure of GRa-1 were analyzed. The results showed that GRa-1 GRb-1 was a neutral polysaccharide and GRb-2 was a basic polysaccharide. HPLC analysis showed that GRa-1 was a homogeneous polysaccharide with a molecular weight of 3 584 Da, while GRb-1 and GRb-2 contained a variety of polysaccharides. The IR spectra of the polysaccharides to be further purified showed that the polysaccharides contained D-type mannofuran sugar. GC-MS showed that the monosaccharides of GRa-1 were composed of arabinose, mannose, glucose and galactose, and the molar ratio was 0.005: 0.014: 0.168: 0.813.3. To study the antioxidant activity and distribution of each component in the separation and purification of polysaccharides. The results show that Crude polysaccharides (GP and GPC) showed strong antioxidant activity in vitro. GRa-1 showed strong activity in Fe2 chelation and O-2 hydroxyl radical scavenging. GRb-1 showed stronger scavenging ability to O-2 and OH. GRb-2 chelates with Fe2 and O-2 with hydroxyl hydroxide with higher scavenging ability. Strong external also has a strong total reduction force. 4. By intraperitoneal injection of glucose tolerance test (Intraperitoneal Glucose Tolerance Test,IPGTT), oral fat tolerance test (Oral Fat Tolerance Test,OFTT), ear swelling test induced by xylene in mice and antioxidation test in aged mice, the antihyperglycemic, antilipids, anti-inflammatory and anti-oxidation activities of GRa-1 were measured. The results showed that GRa-1 could significantly increase the utilization of glucose in mice, accelerate the metabolism of triglycerides in mice, inhibit ear swelling induced by xylene, and enhance the activities of some antioxidant enzymes in serum and liver of aged mice. The content of (MDA) was decreased.
【學(xué)位授予單位】：湖北民族學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TQ464.1

【參考文獻(xiàn)】

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本文編號(hào)：2237145