本文主要研究内容
作者陈光静（2019）在《方竹笋的加工废笋渣中多糖的分离纯化和结构解析及其生物活性研究》一文中研究指出：多糖广泛分布于植物、微生物、动物及水藻中,大量研究证实天然植物多糖具有抗氧化、抗疲劳、抗过敏、抗肿瘤、抗炎、免疫调节、降血糖和益生等诸多生理活性。相较于人工合成药物,植物多糖还具有作用效果相对较好、毒副作用小、安全性高等优点,被认为是合成药物的理想替代品。因此,从新植物原料中寻找活性多糖并对其结构、生物活性等进行分析已逐步成为国内外学者研究的热点。我国竹笋资源丰富,作为世界最大的竹笋生产国和消费国,鲜笋年产量高达500600万吨。由于新鲜竹笋易褐变和木质化,导致其保质期不长,同时,其上市周期较短和集中。因此,大量的鲜笋需要进一步加工保藏如腌制、干制和制作罐头等,但目前加工中存在的普遍问题是竹笋加工过程中原料浪费严重,如笋衣、笋渣和笋头等加工副产物占到了竹笋原料的50%70%,而这部分加工副产物只有少量用做动物饲料,大部分副产物都被丢弃,造成竹笋原料的浪费及环境污染。方竹（Chimonobambusa quadrangularis）因其竹秆呈四棱形略方而得名,是著名的珍贵竹种之一,其竹笋质量好,味道鲜美,深受市场欢迎。然而,方竹笋作为重庆、贵州特有的竹笋资源,在加工过程中也面临大量加工副产物未得到有效利用和开发的问题。因此,提高方竹笋加工副产物的利用率、开发相关的高附加值产品是目前方竹笋加工中急需解决的问题。而目前国内外关于方竹笋多糖的提取、结构解析及生理活性的研究还未见有报道。基于此,本实验以方竹笋加工副产物的废笋渣为研究对象,瞄准其中的多糖活性成分,对方竹笋的加工废笋渣中多糖的提取方法进行研究,同时对比不同提取方法对方竹笋多糖理化性质和体外抗氧化及益生活性的影响,在此基础上对方竹笋多糖进行分离和纯化并对其结构进行解析,再研究方竹笋多糖纯化组分的体外模拟消化和酵解特征,最后研究方竹笋多糖纯化组分对葡聚糖硫酸钠（DSS）诱导的小鼠急性溃疡性肠炎的影响及机理,以期为方竹笋多糖临床药物和保健食品的开发以及方竹笋加工副产物的高附加值转化提供理论依据和实践指导。主要研究结果如下:（1）方竹笋的加工废笋渣中多糖的快速溶剂提取工艺研究采用快速溶剂提取（ASE）技术提取方竹笋的加工废笋渣中的多糖,以粗多糖得率为评价指标,通过单因素和响应面优化实验对其提取工艺参数进行优化。同时与常规热水提取法（HWE）进行对比,考察采用上述两种方法提取得到的方竹笋多糖在得率、主要化学组成、热稳定性和流变学性质方面的差异。实验结果表明,方竹笋的加工废笋渣中多糖快速溶剂提取的最佳提取工艺条件为:笋渣粒径Φ250–380μm,提取温度126°C,循环次数2次,单次循环提取时间22 min。最佳提取条件下,方竹笋粗多糖的得率为9.96±0.39%,显著高于传统热水提取法的得率（7.16±0.32%,p<0.05）,其提取时间远低于传统热水提取法的单次提取时间（4 h）,而两种提取方法提取的方竹笋多糖其总糖含量和热稳定性无显著差异。静态流变学实验结果表明,ASE提取得到的方竹笋多糖（ASE-CPS）和热水提取法提取得到的方竹笋多糖（HWE-CPS）,其溶液都表现出剪切变稀的假塑性流体特征,表观粘度都随着其浓度增加而增大,在相同浓度条件下,ASE-CPS的表观粘度更大;添加CaCl2会降低ASE-CPS和HWE-CPS溶液的表观粘度,添加相同浓度的CaCl2溶液,HWE-CPS溶液的表观粘度降低的更多。动态流变学实验结果表明,HWE-CPS和ASE-CPS为弱凝胶型多糖,添加CaCl2可降低HWE-CPS和ASE-CPS的凝胶强度,且添加相同浓度的CaCl2,HWE-CPS的凝胶强度降低的更多。以上结果说明,与传统热水提取法比较,ASE可作为方竹笋的加工废笋渣中多糖的高效提取方法。（2）不同提取方法对方竹笋多糖理化性质和生物活性影响的研究分别采用快速溶剂提取、热水提取、超声辅助提取、微波辅助提取和复合酶辅助提取五种提取方法提取方竹笋的加工废笋渣中的多糖,对其初步纯化,最终得到5个对应的多糖样品,并对其理化性质进行比较;再采用体外化学抗氧化法综合比较5个多糖样品的氧化自由基吸收能力（ORAC）,清除DPPH自由基、羟基自由基、超氧阴离子自由基和ABTS自由基的能力,金属离子螯合力和还原力;考察5个多糖样品在人工胃液和α-淀粉酶液中的稳定性;将上述5个多糖样品作为碳源,加入到无碳源的MRS基础培养基中,分别接种青春双歧杆菌、两歧双歧杆菌、婴儿双歧杆菌和嗜酸乳杆菌4种益生菌菌种,通过体外厌氧发酵实验,对比不同多糖样品对益生菌的体外增殖效果及产生短链脂肪酸的情况。实验结果表明,不同提取方法显著影响方竹笋多糖的得率和某些理化性质,同时显著影响方竹笋多糖的体外抗氧化活性和益生活性,但对方竹笋多糖在人工胃液和α-淀粉酶液中的稳定性无显著影响,不同提取方法制备的方竹笋多糖在人工胃液和α-淀粉酶液中均表现出较强的稳定性（在人工胃液中水解率<1.3%,在α-淀粉酶液中水解率<5.0%）。理化性质方面,不同提取方法影响方竹笋多糖的得率、化学组成、单糖组成、分子量、粒径大小、Zeta电位和三股螺旋构象。快速溶剂提取法的多糖得率最高,超声辅助提取法制备的方竹笋多糖其糖醛酸含量最高、平均分子量最小、平均粒径最小、Zeta电位的绝对值最大;体外抗氧化活性方面,五种提取方法制备的方竹笋多糖均表现出一定的抗氧化能力,超声辅助提取法制备的方竹笋多糖具有更强的氧化自由基吸收能力,更强的清除DPPH自由基、羟基自由基、超氧阴离子自由基和ABTS自由基能力及更强的Fe2+螯合力和还原力;体外益生活性方面,五种提取方法制备的方竹笋多糖均表现出显著的益生活性,超声辅助提取法和复合酶辅助提取法制备的方竹笋多糖对促进青春双歧杆菌、两歧双歧杆菌、婴儿双歧杆菌和嗜酸乳杆菌体外增殖的效果更佳,能促进上述四种益生菌产生更多的短链脂肪酸。（3）方竹笋的加工废笋渣中多糖的分离纯化、理化性质及结构解析建立DEAE cellulose-52阴离子交换柱和Sephadex G-100葡聚糖凝胶柱两步法分离纯化方竹笋多糖工艺技术路线,对方竹笋废笋渣中多糖进行分离纯化,进一步对方竹笋多糖纯化组分进行纯度鉴定及分子量测定,最后采用化学分析结合仪器分析法对方竹笋多糖纯化组分的结构进行解析。研究结果表明,对方竹笋粗多糖进行分离纯化,最终得到PCPS-1和PCPS-2两个纯化组分,其总糖含量分别为93.26%和92.25%,糖醛酸含量分别为0.28%和0.65%,平均分子量分别为24.58 kDa和123.45 kDa;PCPS-1和PCPS-2均由甘露糖、鼠李糖、葡萄糖醛酸、葡萄糖、半乳糖和阿拉伯糖组成,PCPS-1对应的上述单糖摩尔百分含量分别为0.54%、0.49%、0.33%、5.93%、46.79%和45.93%,PCPS-2对应的上述单糖摩尔百分含量分别为0.46%、1.57%、1.40%、3.36%、45.82%、和47.38%;高碘酸氧化和Smith降解、甲基化分析、气质联机和核磁共振分析结果表明,PCPS-1和PCPS-2均主要由→5)-α-L-Araf-（1→、→3,5）-α-L-Araf-（1→、→6）-β-D-Galp-（1→、→3,6）-β-D-Galp-（1→、T-β-D-Galp、→3）-β-D-Galp-（1→和T-α-D-Glcp七种糖苷键组成,其中PCPS-1对应的上述糖苷键摩尔百分含量分别为37.16%、9.01%、17.38%、14.22%、11.19%、6.08%和4.95%;PCPS-2对应的上述糖苷键摩尔百分含量分别为32.87%、16.81%、16.86%、13.73%、5.91%、10.67%和3.14%;扫描电镜结果表明,PCPS-1主要呈片状结构,表面比较平整,而PCPS-2主要呈棒状结构,表面比较光滑;原子力显微镜和圆二色谱结果显示,PCPS-1和PCPS-2分子在水溶液中均以不规则的聚合体存在;X衍射结果表明,PCPS-1和PCPS-2中只含有少量的微晶结构,主要以无定型形态存在。（4）方竹笋多糖PCPS-2的体外模拟消化及酵解特征的研究采用人体唾液、体外模拟胃液及小肠液消化模型,研究方竹笋多糖纯化组分PCPS-2的体外消化特征,并使用DEAE cellulose-52阴离子交换柱对PCPS-2的消化产物进行纯化,再采用核磁共振技术对其主要糖苷键进行分析,用于比较PCPS-2消化前后主要糖苷键是否发生变化;同时,以人体粪便微菌群为酵解模型,研究方竹笋多糖的体外酵解特征。研究结果表明,PCPS-2在人体唾液、模拟胃液和小肠液中稳定性较好,PCPS-2经人体唾液、模拟胃液和小肠液消化后,其分子量和还原糖含量未发生显著变化,也未释放游离单糖;方竹笋多糖PCPS-2经体外模拟消化后的核磁共振图谱结果表明,PCPS-2经唾液、模拟胃液和小肠液消化后其主要糖苷键并未发生显著变化;体外酵解实验显示,发酵24 h后,PCPS-2中53.36%的总糖被肠道微生物分解利用,发酵液的pH值由8.18降低到5.31,发酵液中α-L-阿拉伯呋喃糖苷酶和β-D-半乳糖苷酶的活性显著增加（p<0.05）,发酵液中乳酸、乙酸、丙酸和总SCFAs的浓度分别从0.64、1.73、0.38和3.11 mmol/L显著增加到8.95、14.03、7.16和31.37 mmol/L。以上结果说明,PCPS-2可被肠道微生物分解利用,能够显著促进肠道微生物产生SCFAs,乳酸、乙酸和丙酸为其主要的SCFAs产物。（5）方竹笋多糖PCPS-2对小鼠急性溃疡性结肠炎的影响及其机理研究采用3%DSS诱导BALB/c雄性小鼠急性溃疡性结肠炎模型,考察方竹笋多糖PCPS-2膳食干预下对小鼠溃疡性结肠炎的作用效果,同时采用酶联免疫技术、qRT-PCR技术和Western blot等技术考察PCPS-2对溃疡性结肠炎小鼠结肠中炎症细胞因子、iNOS和COX-2炎症蛋白、NLRP3炎症小体和NF-κB信号通路的影响,探讨PCPS-2的可能作用机制。研究结果表明,PCPS-2对DSS诱导的小鼠急性溃疡性结肠炎具有显著的保护作用,具体表现为:PCPS-2可显著改善由DSS诱发的小鼠体重减轻、粪便隐血、结肠缩短、结肠水肿与溃疡、脾脏肿大等症状;可有效降低DSS对结肠组织的损伤,结肠炎小鼠结肠粘膜的溃疡得到改善,炎性细胞的浸润减少,隐窝结构得到一定程度恢复。PCPS-2对DSS诱导的小鼠急性溃疡性结肠炎的保护作用主要通过以下几种方式实现:降低结肠炎小鼠结肠中促炎细胞因子TNF-α、IL-1β、IL-6和IL-18的表达量,改善结肠炎小鼠结肠中促炎细胞因子IL-1β、IL-6和IL-18与抗炎细胞因子IL-10间的平衡状态;通过降低结肠炎小鼠血清中NO分泌量、减少结肠组织中MDA含量、提高T-SOD活力、降低MPO活力从而抑制结肠炎小鼠机体的氧化应激反应,修复组织损伤;下调结肠炎小鼠结肠中炎症蛋白iNOS和COX-2的表达;抑制结肠炎小鼠结肠中NLRP3炎症小体的激活;抑制结肠炎小鼠结肠的NF-κB信号通路中p65和IκB-α蛋白的磷酸化。结论:本实验成功建立了方竹笋的加工废笋渣中多糖的快速溶剂提取新工艺,同时建立了DEAE cellulose-52阴离子交换柱和Sephadex G-100葡聚糖凝胶柱两步法分离纯化方竹笋多糖工艺技术路线;证实了提取方法显著影响方竹笋多糖的体外抗氧化活性和益生活性,其中超声辅助提取法制备的方竹笋多糖表现出更显著的体外抗氧化活性和益生活性;方竹笋多糖纯化组分PCPS-1和PCPS-2的主要结构特点为两个纯化组分均主要含→3,5）-α-L-Araf-（1→、→5）-α-L-Araf-（1→、→6）-β-D-Galp-（1→、→3,6）-β-D-Galp-（1→、T-β-D-Galp、→3）-β-D-Galp-(1→和T-α-D-Glcp七种糖苷键;证实了PCPS-2可抵抗人体唾液、模拟胃液和小肠液的消化作用,且经体外模拟消化后其主要糖苷键并未发生显著变化;进一步证明了PCPS-2能够被肠道微生物分解利用,能够显著促进肠道微生物产生乳酸、乙酸和丙酸;最后发现PCPS-2对DSS诱导的小鼠急性溃疡性结肠炎具有显著的保护作用,证明其主要通过降低溃疡性结肠炎小鼠结肠中促炎细胞因子、炎症蛋白iNOS和COX-2的表达,改善结肠炎小鼠机体的氧化应激,抑制结肠炎小鼠结肠中NLRP3炎症小体的激活,抑制结肠炎小鼠结肠的NF-κB信号通路中p65和IκB-α蛋白的磷酸化这几种方式实现对小鼠急性溃疡性结肠炎的保护作用。
Abstract
duo tang an fan fen bu yu zhi wu 、wei sheng wu 、dong wu ji shui zao zhong ,da liang yan jiu zheng shi tian ran zhi wu duo tang ju you kang yang hua 、kang pi lao 、kang guo min 、kang zhong liu 、kang yan 、mian yi diao jie 、jiang xie tang he yi sheng deng zhu duo sheng li huo xing 。xiang jiao yu ren gong ge cheng yao wu ,zhi wu duo tang hai ju you zuo yong xiao guo xiang dui jiao hao 、du fu zuo yong xiao 、an quan xing gao deng you dian ,bei ren wei shi ge cheng yao wu de li xiang ti dai pin 。yin ci ,cong xin zhi wu yuan liao zhong xun zhao huo xing duo tang bing dui ji jie gou 、sheng wu huo xing deng jin hang fen xi yi zhu bu cheng wei guo nei wai xue zhe yan jiu de re dian 。wo guo zhu sun zi yuan feng fu ,zuo wei shi jie zui da de zhu sun sheng chan guo he xiao fei guo ,xian sun nian chan liang gao da 500600mo dun 。you yu xin xian zhu sun yi he bian he mu zhi hua ,dao zhi ji bao zhi ji bu chang ,tong shi ,ji shang shi zhou ji jiao duan he ji zhong 。yin ci ,da liang de xian sun xu yao jin yi bu jia gong bao cang ru a zhi 、gan zhi he zhi zuo guan tou deng ,dan mu qian jia gong zhong cun zai de pu bian wen ti shi zhu sun jia gong guo cheng zhong yuan liao lang fei yan chong ,ru sun yi 、sun zha he sun tou deng jia gong fu chan wu zhan dao le zhu sun yuan liao de 50%70%,er zhe bu fen jia gong fu chan wu zhi you shao liang yong zuo dong wu si liao ,da bu fen fu chan wu dou bei diu qi ,zao cheng zhu sun yuan liao de lang fei ji huan jing wu ran 。fang zhu （Chimonobambusa quadrangularis）yin ji zhu gan cheng si leng xing lve fang er de ming ,shi zhe ming de zhen gui zhu chong zhi yi ,ji zhu sun zhi liang hao ,wei dao xian mei ,shen shou shi chang huan ying 。ran er ,fang zhu sun zuo wei chong qing 、gui zhou te you de zhu sun zi yuan ,zai jia gong guo cheng zhong ye mian lin da liang jia gong fu chan wu wei de dao you xiao li yong he kai fa de wen ti 。yin ci ,di gao fang zhu sun jia gong fu chan wu de li yong lv 、kai fa xiang guan de gao fu jia zhi chan pin shi mu qian fang zhu sun jia gong zhong ji xu jie jue de wen ti 。er mu qian guo nei wai guan yu fang zhu sun duo tang de di qu 、jie gou jie xi ji sheng li huo xing de yan jiu hai wei jian you bao dao 。ji yu ci ,ben shi yan yi fang zhu sun jia gong fu chan wu de fei sun zha wei yan jiu dui xiang ,miao zhun ji zhong de duo tang huo xing cheng fen ,dui fang zhu sun de jia gong fei sun zha zhong duo tang de di qu fang fa jin hang yan jiu ,tong shi dui bi bu tong di qu fang fa dui fang zhu sun duo tang li hua xing zhi he ti wai kang yang hua ji yi sheng huo xing de ying xiang ,zai ci ji chu shang dui fang zhu sun duo tang jin hang fen li he chun hua bing dui ji jie gou jin hang jie xi ,zai yan jiu fang zhu sun duo tang chun hua zu fen de ti wai mo ni xiao hua he jiao jie te zheng ,zui hou yan jiu fang zhu sun duo tang chun hua zu fen dui pu ju tang liu suan na （DSS）you dao de xiao shu ji xing hui yang xing chang yan de ying xiang ji ji li ,yi ji wei fang zhu sun duo tang lin chuang yao wu he bao jian shi pin de kai fa yi ji fang zhu sun jia gong fu chan wu de gao fu jia zhi zhuai hua di gong li lun yi ju he shi jian zhi dao 。zhu yao yan jiu jie guo ru xia :（1）fang zhu sun de jia gong fei sun zha zhong duo tang de kuai su rong ji di qu gong yi yan jiu cai yong kuai su rong ji di qu （ASE）ji shu di qu fang zhu sun de jia gong fei sun zha zhong de duo tang ,yi cu duo tang de lv wei ping jia zhi biao ,tong guo chan yin su he xiang ying mian you hua shi yan dui ji di qu gong yi can shu jin hang you hua 。tong shi yu chang gui re shui di qu fa （HWE）jin hang dui bi ,kao cha cai yong shang shu liang chong fang fa di qu de dao de fang zhu sun duo tang zai de lv 、zhu yao hua xue zu cheng 、re wen ding xing he liu bian xue xing zhi fang mian de cha yi 。shi yan jie guo biao ming ,fang zhu sun de jia gong fei sun zha zhong duo tang kuai su rong ji di qu de zui jia di qu gong yi tiao jian wei :sun zha li jing Φ250–380μm,di qu wen du 126°C,xun huan ci shu 2ci ,chan ci xun huan di qu shi jian 22 min。zui jia di qu tiao jian xia ,fang zhu sun cu duo tang de de lv wei 9.96±0.39%,xian zhe gao yu chuan tong re shui di qu fa de de lv （7.16±0.32%,p<0.05）,ji di qu shi jian yuan di yu chuan tong re shui di qu fa de chan ci di qu shi jian （4 h）,er liang chong di qu fang fa di qu de fang zhu sun duo tang ji zong tang han liang he re wen ding xing mo xian zhe cha yi 。jing tai liu bian xue shi yan jie guo biao ming ,ASEdi qu de dao de fang zhu sun duo tang （ASE-CPS）he re shui di qu fa di qu de dao de fang zhu sun duo tang （HWE-CPS）,ji rong ye dou biao xian chu jian qie bian xi de jia su xing liu ti te zheng ,biao guan nian du dou sui zhao ji nong du zeng jia er zeng da ,zai xiang tong nong du tiao jian xia ,ASE-CPSde biao guan nian du geng da ;tian jia CaCl2hui jiang di ASE-CPShe HWE-CPSrong ye de biao guan nian du ,tian jia xiang tong nong du de CaCl2rong ye ,HWE-CPSrong ye de biao guan nian du jiang di de geng duo 。dong tai liu bian xue shi yan jie guo biao ming ,HWE-CPShe ASE-CPSwei ruo ning jiao xing duo tang ,tian jia CaCl2ke jiang di HWE-CPShe ASE-CPSde ning jiao jiang du ,ju tian jia xiang tong nong du de CaCl2,HWE-CPSde ning jiao jiang du jiang di de geng duo 。yi shang jie guo shui ming ,yu chuan tong re shui di qu fa bi jiao ,ASEke zuo wei fang zhu sun de jia gong fei sun zha zhong duo tang de gao xiao di qu fang fa 。（2）bu tong di qu fang fa dui fang zhu sun duo tang li hua xing zhi he sheng wu huo xing ying xiang de yan jiu fen bie cai yong kuai su rong ji di qu 、re shui di qu 、chao sheng fu zhu di qu 、wei bo fu zhu di qu he fu ge mei fu zhu di qu wu chong di qu fang fa di qu fang zhu sun de jia gong fei sun zha zhong de duo tang ,dui ji chu bu chun hua ,zui zhong de dao 5ge dui ying de duo tang yang pin ,bing dui ji li hua xing zhi jin hang bi jiao ;zai cai yong ti wai hua xue kang yang hua fa zeng ge bi jiao 5ge duo tang yang pin de yang hua zi you ji xi shou neng li （ORAC）,qing chu DPPHzi you ji 、qiang ji zi you ji 、chao yang yin li zi zi you ji he ABTSzi you ji de neng li ,jin shu li zi ao ge li he hai yuan li ;kao cha 5ge duo tang yang pin zai ren gong wei ye he α-dian fen mei ye zhong de wen ding xing ;jiang shang shu 5ge duo tang yang pin zuo wei tan yuan ,jia ru dao mo tan yuan de MRSji chu pei yang ji zhong ,fen bie jie chong qing chun shuang qi gan jun 、liang qi shuang qi gan jun 、ying er shuang qi gan jun he shi suan ru gan jun 4chong yi sheng jun jun chong ,tong guo ti wai ya yang fa jiao shi yan ,dui bi bu tong duo tang yang pin dui yi sheng jun de ti wai zeng shi xiao guo ji chan sheng duan lian zhi fang suan de qing kuang 。shi yan jie guo biao ming ,bu tong di qu fang fa xian zhe ying xiang fang zhu sun duo tang de de lv he mou xie li hua xing zhi ,tong shi xian zhe ying xiang fang zhu sun duo tang de ti wai kang yang hua huo xing he yi sheng huo xing ,dan dui fang zhu sun duo tang zai ren gong wei ye he α-dian fen mei ye zhong de wen ding xing mo xian zhe ying xiang ,bu tong di qu fang fa zhi bei de fang zhu sun duo tang zai ren gong wei ye he α-dian fen mei ye zhong jun biao xian chu jiao jiang de wen ding xing （zai ren gong wei ye zhong shui jie lv <1.3%,zai α-dian fen mei ye zhong shui jie lv <5.0%）。li hua xing zhi fang mian ,bu tong di qu fang fa ying xiang fang zhu sun duo tang de de lv 、hua xue zu cheng 、chan tang zu cheng 、fen zi liang 、li jing da xiao 、Zetadian wei he san gu luo xuan gou xiang 。kuai su rong ji di qu fa de duo tang de lv zui gao ,chao sheng fu zhu di qu fa zhi bei de fang zhu sun duo tang ji tang quan suan han liang zui gao 、ping jun fen zi liang zui xiao 、ping jun li jing zui xiao 、Zetadian wei de jue dui zhi zui da ;ti wai kang yang hua huo xing fang mian ,wu chong di qu fang fa zhi bei de fang zhu sun duo tang jun biao xian chu yi ding de kang yang hua neng li ,chao sheng fu zhu di qu fa zhi bei de fang zhu sun duo tang ju you geng jiang de yang hua zi you ji xi shou neng li ,geng jiang de qing chu DPPHzi you ji 、qiang ji zi you ji 、chao yang yin li zi zi you ji he ABTSzi you ji neng li ji geng jiang de Fe2+ao ge li he hai yuan li ;ti wai yi sheng huo xing fang mian ,wu chong di qu fang fa zhi bei de fang zhu sun duo tang jun biao xian chu xian zhe de yi sheng huo xing ,chao sheng fu zhu di qu fa he fu ge mei fu zhu di qu fa zhi bei de fang zhu sun duo tang dui cu jin qing chun shuang qi gan jun 、liang qi shuang qi gan jun 、ying er shuang qi gan jun he shi suan ru gan jun ti wai zeng shi de xiao guo geng jia ,neng cu jin shang shu si chong yi sheng jun chan sheng geng duo de duan lian zhi fang suan 。（3）fang zhu sun de jia gong fei sun zha zhong duo tang de fen li chun hua 、li hua xing zhi ji jie gou jie xi jian li DEAE cellulose-52yin li zi jiao huan zhu he Sephadex G-100pu ju tang ning jiao zhu liang bu fa fen li chun hua fang zhu sun duo tang gong yi ji shu lu xian ,dui fang zhu sun fei sun zha zhong duo tang jin hang fen li chun hua ,jin yi bu dui fang zhu sun duo tang chun hua zu fen jin hang chun du jian ding ji fen zi liang ce ding ,zui hou cai yong hua xue fen xi jie ge yi qi fen xi fa dui fang zhu sun duo tang chun hua zu fen de jie gou jin hang jie xi 。yan jiu jie guo biao ming ,dui fang zhu sun cu duo tang jin hang fen li chun hua ,zui zhong de dao PCPS-1he PCPS-2liang ge chun hua zu fen ,ji zong tang han liang fen bie wei 93.26%he 92.25%,tang quan suan han liang fen bie wei 0.28%he 0.65%,ping jun fen zi liang fen bie wei 24.58 kDahe 123.45 kDa;PCPS-1he PCPS-2jun you gan lou tang 、shu li tang 、pu tao tang quan suan 、pu tao tang 、ban ru tang he a la bai tang zu cheng ,PCPS-1dui ying de shang shu chan tang ma er bai fen han liang fen bie wei 0.54%、0.49%、0.33%、5.93%、46.79%he 45.93%,PCPS-2dui ying de shang shu chan tang ma er bai fen han liang fen bie wei 0.46%、1.57%、1.40%、3.36%、45.82%、he 47.38%;gao dian suan yang hua he Smithjiang jie 、jia ji hua fen xi 、qi zhi lian ji he he ci gong zhen fen xi jie guo biao ming ,PCPS-1he PCPS-2jun zhu yao you →5)-α-L-Araf-（1→、→3,5）-α-L-Araf-（1→、→6）-β-D-Galp-（1→、→3,6）-β-D-Galp-（1→、T-β-D-Galp、→3）-β-D-Galp-（1→he T-α-D-Glcpqi chong tang gan jian zu cheng ,ji zhong PCPS-1dui ying de shang shu tang gan jian ma er bai fen han liang fen bie wei 37.16%、9.01%、17.38%、14.22%、11.19%、6.08%he 4.95%;PCPS-2dui ying de shang shu tang gan jian ma er bai fen han liang fen bie wei 32.87%、16.81%、16.86%、13.73%、5.91%、10.67%he 3.14%;sao miao dian jing jie guo biao ming ,PCPS-1zhu yao cheng pian zhuang jie gou ,biao mian bi jiao ping zheng ,er PCPS-2zhu yao cheng bang zhuang jie gou ,biao mian bi jiao guang hua ;yuan zi li xian wei jing he yuan er se pu jie guo xian shi ,PCPS-1he PCPS-2fen zi zai shui rong ye zhong jun yi bu gui ze de ju ge ti cun zai ;Xyan she jie guo biao ming ,PCPS-1he PCPS-2zhong zhi han you shao liang de wei jing jie gou ,zhu yao yi mo ding xing xing tai cun zai 。（4）fang zhu sun duo tang PCPS-2de ti wai mo ni xiao hua ji jiao jie te zheng de yan jiu cai yong ren ti tuo ye 、ti wai mo ni wei ye ji xiao chang ye xiao hua mo xing ,yan jiu fang zhu sun duo tang chun hua zu fen PCPS-2de ti wai xiao hua te zheng ,bing shi yong DEAE cellulose-52yin li zi jiao huan zhu dui PCPS-2de xiao hua chan wu jin hang chun hua ,zai cai yong he ci gong zhen ji shu dui ji zhu yao tang gan jian jin hang fen xi ,yong yu bi jiao PCPS-2xiao hua qian hou zhu yao tang gan jian shi fou fa sheng bian hua ;tong shi ,yi ren ti fen bian wei jun qun wei jiao jie mo xing ,yan jiu fang zhu sun duo tang de ti wai jiao jie te zheng 。yan jiu jie guo biao ming ,PCPS-2zai ren ti tuo ye 、mo ni wei ye he xiao chang ye zhong wen ding xing jiao hao ,PCPS-2jing ren ti tuo ye 、mo ni wei ye he xiao chang ye xiao hua hou ,ji fen zi liang he hai yuan tang han liang wei fa sheng xian zhe bian hua ,ye wei shi fang you li chan tang ;fang zhu sun duo tang PCPS-2jing ti wai mo ni xiao hua hou de he ci gong zhen tu pu jie guo biao ming ,PCPS-2jing tuo ye 、mo ni wei ye he xiao chang ye xiao hua hou ji zhu yao tang gan jian bing wei fa sheng xian zhe bian hua ;ti wai jiao jie shi yan xian shi ,fa jiao 24 hhou ,PCPS-2zhong 53.36%de zong tang bei chang dao wei sheng wu fen jie li yong ,fa jiao ye de pHzhi you 8.18jiang di dao 5.31,fa jiao ye zhong α-L-a la bai fu nan tang gan mei he β-D-ban ru tang gan mei de huo xing xian zhe zeng jia （p<0.05）,fa jiao ye zhong ru suan 、yi suan 、bing suan he zong SCFAsde nong du fen bie cong 0.64、1.73、0.38he 3.11 mmol/Lxian zhe zeng jia dao 8.95、14.03、7.16he 31.37 mmol/L。yi shang jie guo shui ming ,PCPS-2ke bei chang dao wei sheng wu fen jie li yong ,neng gou xian zhe cu jin chang dao wei sheng wu chan sheng SCFAs,ru suan 、yi suan he bing suan wei ji zhu yao de SCFAschan wu 。（5）fang zhu sun duo tang PCPS-2dui xiao shu ji xing hui yang xing jie chang yan de ying xiang ji ji ji li yan jiu cai yong 3%DSSyou dao BALB/cxiong xing xiao shu ji xing hui yang xing jie chang yan mo xing ,kao cha fang zhu sun duo tang PCPS-2shan shi gan yu xia dui xiao shu hui yang xing jie chang yan de zuo yong xiao guo ,tong shi cai yong mei lian mian yi ji shu 、qRT-PCRji shu he Western blotdeng ji shu kao cha PCPS-2dui hui yang xing jie chang yan xiao shu jie chang zhong yan zheng xi bao yin zi 、iNOShe COX-2yan zheng dan bai 、NLRP3yan zheng xiao ti he NF-κBxin hao tong lu de ying xiang ,tan tao PCPS-2de ke neng zuo yong ji zhi 。yan jiu jie guo biao ming ,PCPS-2dui DSSyou dao de xiao shu ji xing hui yang xing jie chang yan ju you xian zhe de bao hu zuo yong ,ju ti biao xian wei :PCPS-2ke xian zhe gai shan you DSSyou fa de xiao shu ti chong jian qing 、fen bian yin xie 、jie chang su duan 、jie chang shui zhong yu hui yang 、pi zang zhong da deng zheng zhuang ;ke you xiao jiang di DSSdui jie chang zu zhi de sun shang ,jie chang yan xiao shu jie chang nian mo de hui yang de dao gai shan ,yan xing xi bao de jin run jian shao ,yin wo jie gou de dao yi ding cheng du hui fu 。PCPS-2dui DSSyou dao de xiao shu ji xing hui yang xing jie chang yan de bao hu zuo yong zhu yao tong guo yi xia ji chong fang shi shi xian :jiang di jie chang yan xiao shu jie chang zhong cu yan xi bao yin zi TNF-α、IL-1β、IL-6he IL-18de biao da liang ,gai shan jie chang yan xiao shu jie chang zhong cu yan xi bao yin zi IL-1β、IL-6he IL-18yu kang yan xi bao yin zi IL-10jian de ping heng zhuang tai ;tong guo jiang di jie chang yan xiao shu xie qing zhong NOfen bi liang 、jian shao jie chang zu zhi zhong MDAhan liang 、di gao T-SODhuo li 、jiang di MPOhuo li cong er yi zhi jie chang yan xiao shu ji ti de yang hua ying ji fan ying ,xiu fu zu zhi sun shang ;xia diao jie chang yan xiao shu jie chang zhong yan zheng dan bai iNOShe COX-2de biao da ;yi zhi jie chang yan xiao shu jie chang zhong NLRP3yan zheng xiao ti de ji huo ;yi zhi jie chang yan xiao shu jie chang de NF-κBxin hao tong lu zhong p65he IκB-αdan bai de lin suan hua 。jie lun :ben shi yan cheng gong jian li le fang zhu sun de jia gong fei sun zha zhong duo tang de kuai su rong ji di qu xin gong yi ,tong shi jian li le DEAE cellulose-52yin li zi jiao huan zhu he Sephadex G-100pu ju tang ning jiao zhu liang bu fa fen li chun hua fang zhu sun duo tang gong yi ji shu lu xian ;zheng shi le di qu fang fa xian zhe ying xiang fang zhu sun duo tang de ti wai kang yang hua huo xing he yi sheng huo xing ,ji zhong chao sheng fu zhu di qu fa zhi bei de fang zhu sun duo tang biao xian chu geng xian zhe de ti wai kang yang hua huo xing he yi sheng huo xing ;fang zhu sun duo tang chun hua zu fen PCPS-1he PCPS-2de zhu yao jie gou te dian wei liang ge chun hua zu fen jun zhu yao han →3,5）-α-L-Araf-（1→、→5）-α-L-Araf-（1→、→6）-β-D-Galp-（1→、→3,6）-β-D-Galp-（1→、T-β-D-Galp、→3）-β-D-Galp-(1→he T-α-D-Glcpqi chong tang gan jian ;zheng shi le PCPS-2ke di kang ren ti tuo ye 、mo ni wei ye he xiao chang ye de xiao hua zuo yong ,ju jing ti wai mo ni xiao hua hou ji zhu yao tang gan jian bing wei fa sheng xian zhe bian hua ;jin yi bu zheng ming le PCPS-2neng gou bei chang dao wei sheng wu fen jie li yong ,neng gou xian zhe cu jin chang dao wei sheng wu chan sheng ru suan 、yi suan he bing suan ;zui hou fa xian PCPS-2dui DSSyou dao de xiao shu ji xing hui yang xing jie chang yan ju you xian zhe de bao hu zuo yong ,zheng ming ji zhu yao tong guo jiang di hui yang xing jie chang yan xiao shu jie chang zhong cu yan xi bao yin zi 、yan zheng dan bai iNOShe COX-2de biao da ,gai shan jie chang yan xiao shu ji ti de yang hua ying ji ,yi zhi jie chang yan xiao shu jie chang zhong NLRP3yan zheng xiao ti de ji huo ,yi zhi jie chang yan xiao shu jie chang de NF-κBxin hao tong lu zhong p65he IκB-αdan bai de lin suan hua zhe ji chong fang shi shi xian dui xiao shu ji xing hui yang xing jie chang yan de bao hu zuo yong 。
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