益生菌复合制剂对头孢曲松钠作用小鼠的抗氧化指标、细胞因子及肠道菌群的影响

目的:探究长双歧杆菌BB536和乳双歧杆菌HN019组成的益生菌复合制剂对头孢曲松钠引起的肠道菌群失调的改善效果。方法:头孢曲松钠（2 mg/g）连续灌胃小鼠5 d，构建肠道菌群失调模型小鼠，然后随机分为模型组，益生菌复合制剂低剂量组（2×105 CFU/g）、中剂量组（4×105 CFU/g）、高剂量组（1.2×106 CFU/g），另设正常鼠为对照组，第6 d起各剂量组灌胃相应剂量的益生菌复合制剂，对照组和模型组灌胃等体积的生理盐水，连续灌胃30 d。灌胃结束后无菌收集小鼠粪便对肠道菌群计数，并进行16S rDNA高通量测序分析菌群的多样性以及结构，测定血清中白细胞介素2（Interleukin-2，IL-2）、白细胞介素6（Interleukin-6，IL-6）、白细胞介素1β（Interleukin-1β，IL-1β）和肿瘤坏死因子-α（tumor necrosis factor，TNF-α）水平，测定空肠和肝脏中丙二醛（Malondialdehyde，MDA）、总超氧化物歧化酶（Total Superoxide Dismutase，T-SOD）、谷胱甘肽（glutathione，GSH）和谷胱甘肽过氧化物酶（Glutathione peroxidase，GSH-PX）水平。结果:给药头孢曲松钠后，血清中IL-2、IL-6、IL-1β和TNF-α水平有上升趋势，空肠中MDA水平显著上升（P<0.05）且T-SOD水平显著下降（P<0.05），在高剂量的益生菌复合制剂的干预下，IL-6和IL-1β水平显著降低（P<0.05），IL-2和TNF-α水平极显著降低（P<0.01），此外空肠和肝脏中的MDA水平均明显下降、T-SOD水平显著提高（P<0.05），GSH-PX水平极显著提高（P<0.01），空肠中GSH水平极显著提高（P<0.01）。在肠道微生物方面，抗生素造损后再灌胃益生菌复合制剂，与灌胃益生菌复合制剂前相比小鼠粪便中肠球菌和肠杆菌数量降低，乳杆菌和双歧杆菌数量提高，微生物多样性分析结果表明各剂量组与模型组相比微生物丰富度有所恢复，且中、高剂量组预测的肠道功能与对照组相比更为接近。结论:益生菌复合制剂可以促进抗氧化物质产生，降低细胞因子水平，促进有益菌的繁殖，提高肠道菌群的丰富度，改善了由头孢曲松钠引起的肠道菌群失调状态。

Effects of Probiotic Compound Preparations on Antioxidant Indexes,Cytokines and Intestinal Flora in Mice Treated with Ceftriaxone Sodium

Objective: To explore the effect of a probiotic compound preparation composed of Bifidobacterium longum BB536 and Bifidobacterium lactis HN019 on dysbiosis of murine gut microbiota induced by ceftriaxone sodium. Methods: Ceftriaxone sodium (2 mg/g) was given to mice for 5 days to construct intestinal flora dysbiosis model mice and then they were randomly divided into model group, low-dose (2×105 CFU/g), medium-dose (4×105 CFU/g) and high-dose (1.2×106 CFU/g) groups of probiotic compound preparations. In addition, normal mice were set as the control group. From the 6th day, each dose group was administered with the corresponding dose of probiotic compound preparation, the control group and model group were administered with equal volume of normal saline for 30 days. After the gavage, the mice feces were collected to count the intestinal flora and 16S rDNA high-throughput sequencing was performed to analyze the diversity and structure of the flora. The levels of IL-2, IL-6, IL-1β and TNF-α in serum were measured. The contents of MDA, T-SOD, GSH and GSH-PX in the jejunum and liver were measured. Results: After administration of ceftriaxone sodium, the levels of IL-2, IL-6, IL-1β and TNF-α in serum showed an upward trend, the level of MDA in the jejunum increased significantly (P<0.05) and the level of T-SOD decreased significantly (P<0.05). After the intervention of high-dose probiotic compound preparation, the levels of IL-6 and IL-1β were significantly decreased (P<0.05), the levels of IL-2 and TNF-α were significantly decreased (P<0.01). In addition, the levels of MDA in the jejunum and liver were significantly decreased, the level of T-SOD was significantly increased (P<0.05), the level of GSH-PX was significantly increased (P<0.01) and the level of GSH in the jejunum was significantly increased (P<0.01). In terms of intestinal microbes, the number of Enterococci and Enterobacteria in the feces decreased, while the number of Lactobacilli and Bifidobacteria increased when the probiotic compound preparation was administered after antibiotic injury compared with that before the probiotic compound preparation was administered. The results of microbial diversity analysis showed that the microbial richness of each dose group was restored compared with the model group, and the predicted intestinal function of the middle-dose and high-dose groups was closer to that of the control group. Conclusion: The probiotic compound preparation can promote the production of antioxidants, reduce the level of cytokines, promote the reproduction of beneficial bacteria, increase the abundance of intestinal flora and improve the intestinal flora imbalance caused by ceftriaxone sodium.