【作者】 李杰；

【导师】 郭庆启；

【作者基本信息】 东北林业大学 ， 食品科学与工程， 2025， 硕士

【摘要】 随着现代社会生活节奏的加快、社会压力加剧及生活方式改变等多重因素影响下,在全球,便秘的患病率逐渐增加已达到14%左右,成为影响人类健康的重要因素。便秘会显著影响并降低患者的健康和生活质量,据报道,有便秘症状的患者患其他疾病的风险更大。但是,现如今大多数用于治疗便秘方法会带来一定的副作用。红松种仁油因其含有多种具有药用作用和开发价值的生物活性成分符合“药食同源”健康理念,对于调节肠道功能、缓解便秘和预防胃肠道疾病方面大有裨益。同时为满足人体对脂肪酸平衡和营养的需要以及加强直饮油稳定性,以红松种仁油为基油添加椰子油、米糠油、茶油调配出红松种仁直饮油,研究直饮油缓解便秘的作用和机制。主要研究内容如下:首先对红松种仁直饮油进行制备。使用GC-MS测定红松种仁油及其配方油的脂肪酸组成。根据中国营养学会推荐摄入的脂肪酸比例:饱和脂肪酸(Saturated fatty acid,SFA):单不饱和脂肪酸(Monounsaturated fatty acid,MUFA):多不饱和脂肪酸(Polyunsaturated fatty acid,PUFA)=1:1:1,n-6:n-3=(4-6):1。采用Matlab构建数学模型计算出红松种仁直饮油配方。通过检测直饮油的脂肪酸组成、微量营养成分含量、HS-SPME-GC-MS进行风味分析。验证直饮油的脂肪酸组成、营养价值及风味情况。结果表明直饮油中SFA:MUFA:PUFA=1.14:1.08:1,n-6:n-3=4.17:1,微量营养成分含量为多酚205.30 mg/kg、黄酮92.19 mg/kg、谷维素1053 mg/kg、植物甾醇490.06 mg/kg,直饮油相对含量较高的风味物质是铃兰醛和二烯醛。其次通过小鼠实验研究直饮油缓解小鼠便秘的作用。采用盐酸洛哌丁胺灌胃法建立便秘小鼠模型,以排便量减少、粪便干硬、排便时间增加、毛发粗糙、精神不振为小鼠便秘的判断指标。研究发现与正常小鼠相比便秘小鼠表现出大便干燥、排便困难、神经递质水平异常、氧化应激、结肠组织损伤、occludin与claudin-1的蛋白表达水平显著下降86.11%和25.00%(P<0.05),m RNA表达水平显著下降70.80%和59.00%(P<0.05)。经红松种仁直饮油灌胃后,显著改善了便秘小鼠的排便状况;降低便秘小鼠血清中血管活性肽(VIP),内皮素-1(ET-1)和NO水平以及提高p物质(SP)水平;显著提高便秘小鼠血清中SOD和CAT的含量(P<0.05);结肠组织损伤缓解;显著上调occludin与claudin-1的蛋白表达(P<0.05)以及m RNA的表达水平(P<0.05)。红种仁直饮油通过提高肠道动力、调节血清神经递质、缓解氧化应激、修复肠道屏障损伤、提高紧密连接蛋白的表达以此来缓解小鼠便秘。最后探究红松种仁直饮油缓解小鼠便秘的机制。对小鼠的结肠内容物的肠道菌群和代谢物质进行分析。结果显示便秘小鼠肠道菌群的多样性和丰富度下降,在门水平上便秘小鼠的厚壁菌门(Firmicutes)下降至36.59±2.10%(P>0.05),拟杆菌门(Bacteroidota)上升至51.41±3.91%(P>0.05),厚壁菌门/拟杆菌门(Firmicutes/Bacteroidota,F/B)下降至0.73±0.02(P<0.05),在属水平上Lachnospiraceae＿NK4A136＿group(7.04±0.74%)相对丰度减少。经直饮油灌胃后提高了便秘小鼠厚壁菌门的相对丰度,降低拟杆菌门、疣微菌门和变形菌门的相对丰度,提高了F/B比值,进而恢复了肠道菌群的平衡,在属水平上下调了拟杆菌属和阿克曼菌属相对丰度,显著增加Lachnospiraceae＿NK4A136＿group相对丰度,结果表明直饮油上调了便秘导致的肠道功能紊乱其中高剂量直饮油组肠道菌群功能更接近空白组。在非靶向代谢组学中便秘小鼠的代谢物质发生显著改变,经直饮油灌胃后改善了便秘小鼠的代谢紊乱。KEGG数据库结果表明红松种仁直饮油通过影响叶酸介导的一碳代谢、AMPK信号通路和丙氨酸、天冬氨酸和谷氨酸生物合成代谢途径来改善便秘。更多还原

【Abstract】 With the influence of multiple factors such as the acceleration of the pace of modern life,the aggravation of social pressure and lifestyle changes,the prevalence of constipation has gradually increased by about 14%globally,which has become an important factor affecting human health.Constipation significantly affects and reduces patient health and quality of life,and patients with symptoms of constipation are reported to be at greater risk of other diseases.However,most constipation have side effects.Korean pine seed oil contains a variety of bioactive ingredients with medicinal effects and development value,which is in line with the health concept of"medicine and food",and is of great benefit to regulating intestinal function,relieving constipation and preventing gastrointestinal diseases.At the same time,in order to meet the needs of human body for fatty acid balance and nutrition and strengthen the stability of direct drinking oil,coconut oil,rice bran oil and camellia oil were added with red pine seed oil and mixed with red pine seed oil,and the role and mechanism of direct drinking oil to relieve constipation were studied.The main research contents are described as follows:First,the Korean pine seed seed oil was prepared.The fatty acid composition of Korean pine seed oil and its formula oil was determined using GC-MS.According to the proportion of fatty acids recommended by the Chinese Nutrition Society:saturated fatty acids(SFA):monounsaturated fatty acids(MUFA):polyunsaturated fatty acids(PUFA)=1:1:1,n-6:n-3=(4-6):1.The Matlab was used to calculate the Korean pine seed oil formula.Flavor analysis was performed by detecting the fatty acid composition,trace nutrient content,and HS-SPME-GC-MS.Verify the fatty acid composition,nutritional value and flavor of direct drinking oil.The results showed that SFA:MUFA:MUFA=1.14:1.08:1,n-6:n-3:4.17:1,micronutrient polyphenols 205.30 mg/kg,flavone 92.19 mg/kg,1053 mg/kg,phytosterol 490.06 mg/kg,the relatively high content of oil were bollaldehyde and dioleal.Secondly,the effect of drinking oil directly to relieve constipation in mice.The mouse model of constipation was established by loperamide hydrochloride perfusion method,and reduced defecation volume,dry stool,increased defecation time,rough hair and low spirit were the indicators of constipation in mice.The study found that compared with normal mice,constipated mice showed dry stool,difficult defecation,abnormal neurotransmitter levels,oxidative stress,colon tissue damage,occludin and claudin-1 protein expression levels by86.11%and 25.00%(P<0.05),and m RNA expression levels by 70.80%and 59.00%(P<0.05).After administration,it significantly improved the defecation condition of constipated mice;reduced the vasoactive peptide(VIP)in serum,endothelin-1(ET-1)and NO and increased the level of p substance(SP);significantly increased the amount of SOD and CAT in constipated mice(P<0.05);reduced the amount of colon tissue injury;significantly increased the protein expression of occludin and claudin-1(P<0.05)and the expression level of m RNA(P<0.05).Red seed kernel oil consumption alleviates constipation in mice by improving intestinal motility,regulating serum neurotransmitters,relieving oxidative stress,repairing intestinal barrier damage,and improving the expression of tight junction proteins.Finally,the mechanism of Korean pine seed oil consumption to relieve constipation in mice was explored.The gut microbiota and metabolic materials of the colonic contents of the mice were analyzed.The results showed the decreased diversity and richness of the intestinal microbiota of constipated mice,At the phylum level,the Firmicutes decreased to 36.59±2.10%(P>0.05),Bacteroidetes rose to 51.41±3.91%(P>0.05),Firmicutes/Bacteroidetes(F/B)decreased to 0.73±0.02(P<0.05),The relative abundance of Lachnospiraceae＿NK4A136＿group(7.04±0.74%)was decreased at the genus level.After the direct drinking oil infusion improves the relative abundance of Firmicutes constipation mice,reduce the relative abundance of Bacteroidetes,wart and proteobacteria,improve the F/B ratio,and restore the balance of the intestinal flora,reduce the relative abundance of Bacteroides and ackermania at the genus level,significantly increase the relative abundance of Lachnospiraceae＿NK4A136＿group,the results show that the direct drinking oil upregulated the intestinal dysfunction caused by constipation in the high dose of direct drinking oil group closer to the blank group.Metisms were significantly altered in constipated mice in untargeted metabolomics,and improved metabolic disturbances in constipated mice after direct oil gavage.The KEGG database results indicate that Korean pine seed oil improves constipation by affecting folate-mediated one-carbon metabolism,AMPK signaling pathway and alanine,aspartate and glutamate biosynthesis metabolism pathways.更多还原