约氏乳酸杆菌BS15对小鼠脂肪沉积和非酒精性脂肪肝的影响
发布时间：2018-10-10 18:35
【摘要】：本论文通过饲喂小鼠高脂饲料和灌胃不同浓度的约氏乳酸杆菌BS15,来探讨约氏乳酸杆菌BS15对高脂饲料引起的肥胖和非酒精性脂肪肝的作用,并研究其机理。选用120只雄性ICR小鼠,试验前35d小鼠分为三组,ND组60只小鼠均灌胃磷酸缓冲生理盐水(PBS), L-BS15组和H-BS15组各30只分别灌胃不同浓度约氏乳酸杆菌BS15(2×107或2×108 cfu/d),均给予普通饲料(NCD)。上述处理35d后将ND组小鼠分出一半,分出小鼠饲喂高脂饲料和灌胃PBS为HFD组,其余小鼠继续饲喂NCD和灌胃PBS仍为ND组；L-BS15组和H-BS15组小鼠改喂高脂饲料同时继续灌胃不同浓度约氏乳酸杆菌BS15,持续12周。在第35、63、91、119d采取小鼠样本进行检测,试验结果如下：1.与ND组小鼠相比,HFD组体重显著升高；与HFD组相比,L-BS15和H-BS15组小鼠体重显著降低。2.肝脏病理组织学切片观察,HFD组小鼠肝脏内空泡化严重,肝脏脂肪变性较L-BS15和H-BS15组显著。3.通过流式细胞技术(FCM)对小鼠肝脏细胞进行检测,与ND组相比,HFD小鼠肝脏细胞凋亡率显著升高；L-BS15和H-BS15组肝脏细胞凋亡率较HFD组显著降低。4.实时荧光定量(RT-PCR)技术检测小鼠盲肠内容物特定菌群发现,BS15的补充显著增加了肠道内乳酸菌、约氏乳酸杆菌及拟杆菌的数量,同时降低了肠道内肠杆菌和厚壁杆菌门／拟杆菌门的比例,有效改善了高脂饲料诱导的肠道菌群的紊乱。5.实时荧光定量(RT-PCR)技术检测小鼠肝脏内脂肪代谢相关因子(ACC1、 FAS、FIAF、PPARy)基因表达发现,与ND组相比,HFD组ACC1、FAS和PPARy基因表达量显著升高,FIAF表达量显著降低；灌胃BS15后,ACC1、FAS和PPARγ基因表达量较HFD组显著降低,FIAF表达量显著升高。6.肠道通透性试验表明,高脂饲料显著诱导了HFD组小鼠肠道通透性；BS15的补充显著降低了小鼠肠道通透性。7.机体炎症相关指标检测表明,HFD组小鼠血清中LPS含量和肝脏组织TNF-α表达水平在119 d显著高于ND组。灌胃BS15后,与HFD组相比,血清LPS含量和肝脏TNF-α表达量在L-BS15组和H-BS15组显著降低。与ND组相比,从第63d到119d小鼠血清中C反应蛋白(CR_P)含量在HFD组显著升高。灌胃BS15后,与HFD组相比,从第63d到119d小鼠血清中CRP含量在L-BS15组和H-BS15组显著降低。BS15显著降低了高脂饲料诱导的机体炎症。8.小鼠胰岛素抗性检测发现,与ND组相比在第63、91、119 d, HFD组小鼠血清中胰岛素含量和胰岛素抗性均显著升高,灌胃BS15后,小鼠血清中胰岛素含量及胰岛素抗性在63、91、119d三个检测点均显著降低。与ND组相比,小鼠血清中TG、TC、LDL-C、ALT、FFA含量在HFD组显著升高,HDL-C含量在HFD组显著降低。灌胃BS15后,与HFD组相比,TG、LDL-C、ALT、FFA含量显著降低,HDL-C含量显著升高。BS15的补充抑制了胰岛素抗性的产生及血脂指标的紊乱。9.氧化应激相关指标检测发现,与ND组相比HFD组肝脏线粒体和肝细胞胞浆中H2O2、GSH和MDA含量均显著升高,CAT、GSH-PX和T-SOD酶活力在线粒体和胞浆中显著降低。灌胃BS15后与HFD组相比,L-BS15组和H-BS15组线粒体内H2O2、GSH和MDA含量均显著降低,CAT和GSH-PX酶活力显著升高,T-SOD酶活力变化不显著。与HFD组相比,L-BS15组和H-BS15组胞浆内H2O2、GSH含量均显著降低,MDA、T-SOD酶活力显著升高,CAT、GSH-PX酶活力仅在H-BS15组内升高显著。BS15的补充有利于抑制肝脏组织氧化应激的发生。综上所述,约氏乳酸杆菌BS15具有抗肥胖和预防非酒精性脂肪肝的作用,其主要机理是通过改善肠道菌群,降低肠道通透性、机体炎症、肝脏氧化应激和胰岛素抵抗。
[Abstract]:In order to investigate the effect of Lactobacillus casei BS15 on obesity and non-alcoholic fatty liver induced by high fat feed, the mechanism was studied by feeding mice with high fat feed and Lactobacillus bulgaricus BS15 at different concentrations. A total of 120 male ICR mice were divided into three groups, and 60 mice in ND group were given oral phosphate buffered saline (PBS), and 30 rats in L-B15 group and H-BS15 group were respectively given different concentrations of Lactobacillus bulgaricus BS15 (2, 107 or 2, 108 cfu/ d), all of which were given normal feed (NCD). After 35d treatment, the ND group mice were divided into half, the mice were fed with high fat feed and PBS as HFD group, the remaining mice continued to feed NCD and gavage PBS were still ND group; L-BS15 group and H-BS15 group mice were fed with high fat feed while continuing to gavage at different concentrations of Lactobacillus bulgaricus B15 for 12 weeks. Mice samples were tested on Days 35, 63, 91, 119d and the results were as follows: 1. Compared with the ND group, the body weight of the HFD group increased significantly; compared with the HFD group, the body weight of the L-B15 and H-B15 groups decreased significantly. The pathological histological sections of liver showed that the vacuolization of liver was severe in HFD group, and the fatty degeneration of liver was higher than that in L-BS15 and H-BS15 group. The liver cells of mice were detected by flow cytometry (FCM). Compared with ND group, the apoptosis rate of liver cells in HFD mice increased significantly, and the apoptosis rate of liver cells in L-B15 and H-B15 groups was significantly lower than that in the HFD group. The real-time fluorescence quantitative (RT-PCR) technique is used for detecting the specific flora of the cecum contents of the mice, and the supplementation of the B15 remarkably increases the number of lactic acid bacteria in the intestinal tract, the lactobacilli and the pseudo-bacillus, and simultaneously reduces the proportion of the enterobacteriaceae and the thick-walled bacillus door/ planned bacillus door in the intestinal tract, effectively improving the disorder of intestinal flora induced by high fat feed. The expression of ACC1, FAS, FIAF, PPARy in liver of mice was detected by real-time fluorescence quantitative (RT-PCR) technique. Compared with the HFD group, the expression level of the FAS and Jurkat gene was significantly decreased, and the expression level of FIF increased significantly. The intestinal permeability test showed that the high fat diet significantly induced intestinal permeability in the HFD group, and the supplementation of B15 significantly reduced the intestinal permeability of mice. The results showed that the expression level of LPS in serum of HFD group and liver tissue was significantly higher than that of ND group in HFD group. Compared with the HFD group, the expression levels of LPS and liver TNF-B15 decreased significantly in the L-B15 group and the H-B15 group compared with the HFD group. Compared with ND group, the content of C-reactive protein (CR _ P) in serum of mice from 63d to 119d increased significantly in the HFD group. Compared with the HFD group, the CRP levels in the serum of mice from 63d to 119d were significantly lower in the L-B15 group and in the H-B15 group than in the HFD group after gavage of B15. BS15 significantly reduced body inflammation induced by high fat feed. Compared with ND group, the insulin content and insulin resistance in serum of mice were significantly increased compared with ND group, and the insulin content and insulin resistance in serum of mice were significantly decreased at 63, 91 and 119d after intragastric administration of B15. Compared with ND group, the content of TG, TC, LDL-C, ALT and HDL in serum of mice increased significantly in HFD group, and HDL-C content decreased significantly in HFD group. After intragastric administration of BS15, TG, LDL-C, ALT, HDL-C content were significantly decreased compared with HFD group, and HDL-C content increased significantly. The addition of B15 inhibited the generation of insulin resistance and disorder of blood lipid index. Compared with ND group, the content of H2O2, GSH and MDA in liver mitochondria and liver cell cytoplasm of HFD group increased significantly compared with ND group, CAT, GSH-PX and T-SOD activity decreased significantly in mitochondria and cytoplasm. Compared with HFD group, the contents of H2O2, GSH and MDA in the mitochondria of L-BS15 and H-BS15 groups were significantly decreased compared with the HFD group, but the activity of CAT and GSH-PX increased significantly, and the activity of T-SOD was not significant. Compared with HFD group, the content of H2O2 and GSH in plasma of L-BS15 group and H-BS15 group decreased significantly, MDA, T-SOD activity increased significantly, CAT, GSH-PX activity increased significantly in H-BS15 group. The supplementation of B15 is beneficial to inhibit the occurrence of oxidative stress in liver tissues. In conclusion, Lactobacillus casei BS15 has the function of resisting obesity and preventing non-alcoholic fatty liver, and its main mechanism is to improve intestinal flora, reduce intestinal permeability, body inflammation, liver oxidative stress and insulin resistance.
【学位授予单位】：四川农业大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.5;R589.2
本文编号：2262814
[Abstract]:In order to investigate the effect of Lactobacillus casei BS15 on obesity and non-alcoholic fatty liver induced by high fat feed, the mechanism was studied by feeding mice with high fat feed and Lactobacillus bulgaricus BS15 at different concentrations. A total of 120 male ICR mice were divided into three groups, and 60 mice in ND group were given oral phosphate buffered saline (PBS), and 30 rats in L-B15 group and H-BS15 group were respectively given different concentrations of Lactobacillus bulgaricus BS15 (2, 107 or 2, 108 cfu/ d), all of which were given normal feed (NCD). After 35d treatment, the ND group mice were divided into half, the mice were fed with high fat feed and PBS as HFD group, the remaining mice continued to feed NCD and gavage PBS were still ND group; L-BS15 group and H-BS15 group mice were fed with high fat feed while continuing to gavage at different concentrations of Lactobacillus bulgaricus B15 for 12 weeks. Mice samples were tested on Days 35, 63, 91, 119d and the results were as follows: 1. Compared with the ND group, the body weight of the HFD group increased significantly; compared with the HFD group, the body weight of the L-B15 and H-B15 groups decreased significantly. The pathological histological sections of liver showed that the vacuolization of liver was severe in HFD group, and the fatty degeneration of liver was higher than that in L-BS15 and H-BS15 group. The liver cells of mice were detected by flow cytometry (FCM). Compared with ND group, the apoptosis rate of liver cells in HFD mice increased significantly, and the apoptosis rate of liver cells in L-B15 and H-B15 groups was significantly lower than that in the HFD group. The real-time fluorescence quantitative (RT-PCR) technique is used for detecting the specific flora of the cecum contents of the mice, and the supplementation of the B15 remarkably increases the number of lactic acid bacteria in the intestinal tract, the lactobacilli and the pseudo-bacillus, and simultaneously reduces the proportion of the enterobacteriaceae and the thick-walled bacillus door/ planned bacillus door in the intestinal tract, effectively improving the disorder of intestinal flora induced by high fat feed. The expression of ACC1, FAS, FIAF, PPARy in liver of mice was detected by real-time fluorescence quantitative (RT-PCR) technique. Compared with the HFD group, the expression level of the FAS and Jurkat gene was significantly decreased, and the expression level of FIF increased significantly. The intestinal permeability test showed that the high fat diet significantly induced intestinal permeability in the HFD group, and the supplementation of B15 significantly reduced the intestinal permeability of mice. The results showed that the expression level of LPS in serum of HFD group and liver tissue was significantly higher than that of ND group in HFD group. Compared with the HFD group, the expression levels of LPS and liver TNF-B15 decreased significantly in the L-B15 group and the H-B15 group compared with the HFD group. Compared with ND group, the content of C-reactive protein (CR _ P) in serum of mice from 63d to 119d increased significantly in the HFD group. Compared with the HFD group, the CRP levels in the serum of mice from 63d to 119d were significantly lower in the L-B15 group and in the H-B15 group than in the HFD group after gavage of B15. BS15 significantly reduced body inflammation induced by high fat feed. Compared with ND group, the insulin content and insulin resistance in serum of mice were significantly increased compared with ND group, and the insulin content and insulin resistance in serum of mice were significantly decreased at 63, 91 and 119d after intragastric administration of B15. Compared with ND group, the content of TG, TC, LDL-C, ALT and HDL in serum of mice increased significantly in HFD group, and HDL-C content decreased significantly in HFD group. After intragastric administration of BS15, TG, LDL-C, ALT, HDL-C content were significantly decreased compared with HFD group, and HDL-C content increased significantly. The addition of B15 inhibited the generation of insulin resistance and disorder of blood lipid index. Compared with ND group, the content of H2O2, GSH and MDA in liver mitochondria and liver cell cytoplasm of HFD group increased significantly compared with ND group, CAT, GSH-PX and T-SOD activity decreased significantly in mitochondria and cytoplasm. Compared with HFD group, the contents of H2O2, GSH and MDA in the mitochondria of L-BS15 and H-BS15 groups were significantly decreased compared with the HFD group, but the activity of CAT and GSH-PX increased significantly, and the activity of T-SOD was not significant. Compared with HFD group, the content of H2O2 and GSH in plasma of L-BS15 group and H-BS15 group decreased significantly, MDA, T-SOD activity increased significantly, CAT, GSH-PX activity increased significantly in H-BS15 group. The supplementation of B15 is beneficial to inhibit the occurrence of oxidative stress in liver tissues. In conclusion, Lactobacillus casei BS15 has the function of resisting obesity and preventing non-alcoholic fatty liver, and its main mechanism is to improve intestinal flora, reduce intestinal permeability, body inflammation, liver oxidative stress and insulin resistance.
【学位授予单位】：四川农业大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.5;R589.2
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1 辛金鸽;约氏乳酸杆菌BS15对小鼠脂肪沉积和非酒精性脂肪肝的影响[D];四川农业大学;2014年
，本文编号：2262814
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