生姜多糖的提取及其对糖尿病小鼠肠道菌群的调节作用

汪妮; 陈梦霞; 孟凡强; 周立邦; 陆兆新

doi:10.13386/j.issn1002-0306.2022060155

生姜多糖的提取及其对糖尿病小鼠肠道菌群的调节作用

Extraction of Polysaccharide from Zingiber officinale Roscoe and Its Regulatory Effect on Intestinal Flora in Diabetic Mice

摘要

摘要: 目的：研究生姜多糖对高脂饮食（High-fat diet，HFD）和小剂量链脲佐菌素（Streptozotocin，STZ）联合诱导的2型糖尿病（Type 2 diabetes，T2D）小鼠肠道菌群的调节作用。方法：采用热水浸提乙醇沉淀的方法，从生姜中提取出多糖，通过单因素实验和正交试验优化提取条件，随后选择最佳的脱蛋白方法得到生姜多糖（Ginger polysaccharide，GP），并通过测定GP对2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸二铵盐（2,2'-Azinobis-（3-ethylbenzthiazoline-6-sulphonate），ABTS）自由基和羟基自由基的清除能力，评价了其体外抗氧化能力。随后，通过高脂饮食和STZ建立2型糖尿病小鼠模型，测定各组小鼠的空腹血糖（Fasting blood glucose，FBG）浓度和糖化血清蛋白（Glycated serum protein，GSP）水平，采用16S rRNA高通量测序技术分析糖尿病小鼠的肠道菌群组成。结果：GP最优提取工艺为料液比1:20 g/mL、提取温度90 ℃、提取时间1 h；采用Sevag试剂处理5次进行脱蛋白得到GP，得率为2.91%±0.25%，其中GP中总糖含量为43.44%±0.99%；当GP浓度达到4 mg/mL时，GP对ABTS^＋自由基和羟基自由基的清除率分别为66.09%和65.73%；在GP的干预下，显著降低了糖尿病小鼠FBG浓度和GSP水平（P<0.05），16S rRNA测序结果表明，GP改变了糖尿病小鼠的肠道菌群组成和相对丰度，增加了阿德勒克氏菌属（Adlercreutzia）、阿克曼菌属（Akkermansia）和乳酸杆菌属（Lactobacillus）等有益菌的相对丰度，降低了普雷沃氏菌属（Prevotella）等有害菌的相对丰度。结论：通过热水浸提法提取和Sevag脱蛋白所得生姜多糖能改变糖尿病小鼠肠道菌群的组成，该研究结果为生姜多糖预防和治疗T2D提供了理论依据。

Abstract: Objectives: To investigate the regulatory effect of ginger polysaccharides on the intestinal flora of mice with type 2 diabetes (T2D) induced by the combination of a high-fat diet (HFD) and low-dose streptozotocin (STZ). Methods: Polysaccharides were extracted from ginger by hot water extraction and ethanol precipitation. Single-factor experiments and an orthogonal experiment were used to optimize the extraction conditions, and then the best deproteinization method was selected to obtain ginger polysaccharide (GP). The scavenging ability of GP to 2,2'-diazo-bis-3-ethylbenzothiazoline-6-sulfonic diammonium salt (ABTS⁺) free radicals and hydroxyl radicals evaluated its antioxidant capacity in vitro. Subsequently, a type 2 diabetes mouse model was established by a high-fat diet and STZ. The fasting blood glucose (FBG) concentration and glycated serum protein (GSP) level of mice in each group were measured. The composition of the intestinal flora of diabetic mice was analyzed by 16S rRNA high-throughput sequencing technique. Results: The optimum extraction process of GP was as follows: The ratio of material to water was 1:20 g/mL, the extraction temperature was 90 ℃, the extraction time was 1 h, and Sevag reagent was used 5 times to deproteinize to obtain GP, the yield was 2.91%±0.25%, of which the total sugar content was 43.44%±0.99%. At the concentration of 4 mg/mL in GP, the scavenging rates of ABTS⁺ free radical and hydroxyl free radical by GP were 66.09% and 65.73% respectively. After the intervention of GP, FBG concentration and GSP level in the diabetic mice were significantly reduced (P<0.05). The 16S rRNA sequencing results showed that GP altered the gut microbiota composition and relative abundance in diabetic mice, and the relative abundance of beneficial bacteria such as Adlercreutzia, Akkermansia and Lactobacillus increased, while the relative abundance of harmful bacteria such as Prevotella decreased. Conclusions: Ginger polysaccharides obtained by hot water extraction and Sevag deproteinization could alter the composition of the intestinal flora in diabetic mice. The results of this study would provide a theoretical basis for ginger polysaccharide in the prevention and treatment of T2D.