Research For Effects Of Complex Probiotics On High Fat/Sugar Diet Induced Obesity Mice And Their Offspring Lipid Metabolism

Objective:To analyze effects of complex probiotics on the lipid metabolism of obesity mice and their offspring.Methods: 1.30 female C57BL/6J mice were randomly devided into control group,obesity group and microbiota intervention group,with 10 mice in each group.Each group was fed with standard diet,high fat/sugar diet and high fat/sugar diet added with complex probiotics(Bacillus bifidus,Streptococcus thermophilus,lactobacillus casei and Lactobacillus rhamnosus GG),respectively.The simple feeding period was six weeks.After the obesity mice model were established,female mice were mated with male mice.The pregnancy period was about three weeks.During the experimental period,all the mice were fed with their own group diet.All the pups were fed with breast milk.Three weeks later,all the mice were sacrificed.2.Maternal mice body weight was measured every week.Stool and blood samples of maternal mice in lactation 21 day were collected for microbiota and serum lipid analyses.Liver sample of maternal mice in lactation 21 day was collected for HE staining analysis.3.Pups bodyweight in postnatal 3,7,14 and 21 day were measured.Stool and blood samples of pups in postnatal 21 day were collected for microbiota and serum lipid analyses.Liver sample of maternal mice in lactation 21 day was collected for HE staining analysis.4.Stool microbiota analysis:Microbial population DNA was extracted from maternal mice and their pups stool.After PCR amplification,DGGE analysis was used.Differential bands were cut and reclaimed for sequencing analysis.5.Serum lipid detection:Total cholesterol(TC),triglyceride(TG),high density lipoprotein(HDL)and low density lipoprotein(LDL)were examined using ELISA.The ratio of TC/HDL and LDL/HDL were calculated.Results: 1.The effect for complex probiotics intervention on the weight of maternal mice The weight of maternal mice was measured every week to draw a change curve.In the whole period of breeding,at every time point,the weight of mice in the obesity group which were fed with high fat/sugar diet was higher than that of the control group.However,at every time point,the weight of maternal mice in the microbiota intervention group was lower than that of the obesity group,and was similar with the control group.2.The effect for complex probiotics intervention on the intestinal microbiota of maternal mice Cluster analysis showed that the mice in the control group were clustered into one group,and the mice in the obesity group were clustered into another group.The results showed that there was an obvious difference between the intestinal microbiota of obesity maternal mice and control maternal mice.The cluster analysis of the probiotics intervention group and the control group were mixed,and there was no obvious clustering phenomenon.This indicated that the intestinal flora formulation of complex probiotics intervened maternal mice were similar to that of the control maternal mice.3.The effect for complex probiotics intervention on the serum lipid of maternal mice Compared with the control group,the serum lipid detection of the maternal mice in obesity group showed that total cholesterol(TC),low density lipoprotein(LDL)and the ratio of TC/HDL and LDL/HDL were obviously increased.The result of high density lipoprotein(HDL)of the maternal mice of the obesity group was obviously lower than that of the control group.However,there was no significant difference of triglyceride(TG)level between the obesity group and the control group.After microbiota intervention the levels of TC and LDL,ratios of TC/HDL and LDL/HDL were obviously lower than those of the obesity group,the serum level of HDL was obviously increased and there was no significant change of TG.Compared with the control group,the levels of TC,LDL,TC/HDL and LDL/HDL in microbiota intervention group was still increased,however,there were no significant changes in TG and HDL.4.The effect for complex probiotics intervention on the liver histology of maternal mice After HE staining,the fat-like denaturation were showed in liver paraffin section of maternal mice under the microscope which was led by high fat /sugar diet.After intervention with probiotics,the fat-like denaturation of maternal mice liver was reduced.5.The body weight curve of pups At each detection points,the weight of pups in the obesity group was obvious higher than that of the pups in control group.The weight of the pups in the microbiota intervention group was lower than that of the pups in the obesity group,but similar to that of the control group.6.The DGGE analysis for pups stool microbiota The cluster analysis showed that the obesity group pups were clustered into one group,and the control group and the microbiota intervention group pups were clustered into one big group.This indicated that there were obvious differences between the intestinal microbiota of the obesity group and the control group,but there was no difference between the microbiota intervention group and the control group for the intestinal flora formation.7.The serum lipid analysis for pups Compared with the control group,the levels of TC,HDL,LDL,TC/HDL and LDL/HDL were obviously increased in the obesity group pups,and there was no obviously change in TG.Compared with the obesity group,the levels of TC,HDL,LDL,TC/HDL and LDL/HDL were obviously decreased in the microbiota intervention group pups.Compared with the control group,the changes of serum lipid level was not significant in microbiota intervention group.8.The liver histology changes for pups Under the microscope,the fat-like denaturation in the liver of obesity group pups was showed.The liver morphous of microbiota intervention group pups was similar to that of the control group.Conclusions: 1.Hight fat /sugar diet induced increase of the body weight,changes of inteastinal microbiota,disorder of serum lipid metabolism and liver fat-like denaturation in mice.2.Microbiota intervention could decrease female mice body weight,improve intestinal microbiota and serum lipid metabolism disorder and alleviate the liver fat-like denaturation.3.Hight fat /sugar diet induced increase of the body weight,changes of inteastinal microbiota,disorder of serum lipid metabolism and liver fat-like denaturation in maternal obesity pups 4.Microbiota intervention could decrease body weight,improve intestinal microbiota and serum lipid metabolism disorder and alleviate the liver fat-like denaturation in maternal hight fat /sugar diet pups.