Effects Of Cysteine ​​and Glutamine On Intestinal Flora And Antioxidant Index In Rats

Intestinal flora is the basis of maintaining intestinal and normal immune functions of animal gut, which has the vital significance to the health of the host. High fat content diet can make the body produce excess free radicals and oxidative stress, and cause dyslipidemia and disorders of intestinal flora, and then seriously impact the health and performance of animals. This study based on in vitro fermentation experiments was conducted to establish oxdative stress model by adding appropriate amount of H2O2 in modified basal medium of GAM, and to investigate the effect of Arginin(Arg), Cysteine(Cys) and Glutamine(Gln) on content of Lactic acid bacteria and e. coli bacteria in the fermented liquid. And then the study would researched the effects of supplemental Cys and Gln in rats’ diets on the main bacteria content of intestinal, part indexes of blood and antioxidant, and fat content of body. That would provide theory basis to further study the intestinal flora and the functional amino acids.Experiment design included four parts. The first part was fermentation test in vitro of the whole intestinal flora. The test was divided into 7 groups included control group and 6experiment groups, and the test groups were repectively added 0.6% and 1.2% of Arg, Cys and Gln in, and eatch GAM base medium was 35 mL. Each group set up three replicates. Its pH and amounts of lactic acid bacteria, bifidobacterium, E. coli, Butyric acid bacteria was detected on0,6,12,24 h after fermentation, respectively. The objective of second part is to construct the oxidative stress model. The test was divided into six groups include control group and 5experiment groups, and the experiment groups was added 0.5, 1, 2, 4, 5 mM of H2O2 in GAM basal medium, respectively. Its amounds of lactic acid bacteria and E. Coli in fermented liquid was respectively detected on 0, 6, 12, 24 h after fermentation. Then the appropriate content of H2O2 was determined. The objective of the third part, under oxidative stress mode, was to determine the effect of Arg, Cys and Gln on the content of lactic acid bacteria and E. Coli in fermented liquid. The test was divided into 8 groups, and each group set up three replicates. The8 groups were control group(the whole intestinal bacteria media,GAM), the negative control group(the whole intestinal bacteria media+2 mM H2O2), and other groups respectively added0.6%, 1.2% Arg, Cys and Gln in bacteria media of negative control group. And then the significant treatment screened would use to subsequent experiment. The experiment of fourth part was the effect of Gln and Cys on intestinal major bacterial contents, blood indexes, and antioxidant indexes in tissue. Sixty rats(30 male and 30 female) with simialar weeks(8-12weeks) and weight(240-350 kg) were selected and divided into 6 groups with randomized experiment design. Test groups were control group of normal diets, high fat diets group and other groups included respectively added add 0.6%, 1.2% Gln and Cys in high fat diets. The results show that:1. The effects of adding different concertrations of Arg, Cys and Gln in culture medium of rat feces on the pH and content of lactic acid bacteria, bifidobacteria, E. coli, and butyric acid bacteria were significant(P<0.05).2. The optimum H2O2 concentration of constructed the oxidative stress model in vitro was 2mM.3. Compared with control group of GAM, The content of lactic acid bacteria and E. coli in ferment liquids were all significant decreased after adding 2 mM H2O2 in basal GAM medium(P<0.05); The lactic acid bacteria and E. coli content of test groups added different concentrations of Arg, Cys and Gln were all higher than H2O2 group, but there was no difference between test group and control group of GAM(P>0.05).4. The amound of Lactic acid bacteria and bifidobacterium in rat any segment intestinal of high fat diet group were lower than normal diet group(P <0.05), while the amound of E. coli and butyric acid bacteria were incresed significantly. The amound of four bacteria could recover normal levels by adding Cys and Gln.5. Compared with normal group, ROS、SOD、MDA、T-AOC levels in rat blood and intestinal tissue increased significantly(P<0.05), and then decreased significantly(P<0.05) with adding Cys and Gln, while the difference between test group of Cys and Gln and normal group was not significant(P>0.05). ROS、SOD、MDA、T-AOC levels of that decreased more obvious with the increasing concentrations of Cys and Gln.6. The content of bacterial endotoxin in spleen and Peyer’s patch of high fat diets group were all higher than normal group(P<0.05), and that content of added 1.2% Cys in high fat diets was significant decreased(P<0.05) and was closed normal group levels.7. The TG, TC, LDL-C levels of high fat diets group were higher than normal group(P<0.05),which caused hyperlipidemia. The addition of Cys and Gln significantly decreased tha levels of of high fat diets group, experimental group was significantly lower than the targets fat group(P<0.05), the effect of Cys group was better.8. Compared with high fat diets group, the fat content of Cys and Gln group decreased significantly(P<0.05), and the levels of best decreased was Cys group. but there was no differecne between the group of Cys and Gln and control group(P>0.05).The comprehensive results indicated that: the supplementation of Cys and Gln in high fat diets could increase the levels of Lactobacillus and bifidobacterium, decrease E. coli levels in intestinal, increase Antioxidant indicators of ROS、SOD、MDA、T-AOC et al. of blood and intestinal tissue, decrease TG、TC、LDL-C levels of blood, and reduce the concentration of bacterial endotoxin of spleen and peyer’s patch. And then realized objectives of decreasing body fat, recovering all indicators, and eliminating oxidative stress.