The Effect And Mechanism Research Of Intestinal Probiotics On Osteoporosis

Aims The microbes play an important role to physiological activities of human.The largest micro ecosystem in the human body is the intestinal micro ecology system.Gut microbes are enormous and various.The genomes of gut microbiota are 150 times of the human.In recent years,gut microbiota has been seen as a very important organ of human.So gut microbiota is closely associated with health.Under normal circumstances,the intestinal micro ecology systems will remain relatively stable state,this kind of dynamic balance is good for the human body.Once the balance was broken,a series of disease,including a variety of inflammatory bowel disease,obesity,diabetes,asthma,osteoporosis,and the metabolic syndrome would be caused.Osteoporosis is one of the most closely relationship diseases with the elderly.Bone loss and degradation of bone tissue structure are the characteristics of osteoporosis.The greatest harm is osteoporosis fracture.In recent years,the studies found that gut microbiota can cause osteoporosis.These microbes may affect activity of osteoclasts and osteoblasts by its metabolites or changing metabolism and the immune system of host through,to affect bone metabolism.Now,cell researches of the relationship of intestinal probiotics and bone metabolism have only a small number.We analyse the changes of gut microbiota in patients with osteoporosis,and use metabolites of two kinds of probiotics(lactobacillus acidophilus and LGG),to research the effect of MC3-T3 and RAW 264.7 cell.This study will research the relationship between gut microbiota and osteoporosis,explore the mechanism of the change of bone metabolism by gut microbiota through bone formation and bone resorption.Methods We selected 8 patients with osteoporosis from Tianjin Medical University General Hospital,department of geriatrics,between 81~97 years old(include 4 men and 4 women).And selected 7 patients without osteoporosis and other metabolic diseases,regarded them as the control group.Test the blood biochemical index and bone turnover markers of all patients from two groups,and analyze fecal microbiota by 16 S rRNA Sequencing.Firstly,we added the supernatant of two kinds of probiotics and butyrate to culture medium.Secondly,we induced MC3-T3 cell to osteoblasts by ?-sodium glycerophosphate,and induced RAW 264.7 cell to osteoclasts by RANKL.Thirdly,we used MTT to test the proliferation and differentiation of osteoclasts and osteoblasts;test the maturity and the mineralization of osteoblast by ALP and alizarin red staining;test the maturity and differentiation of osteoclasts by TRAP staining.Finally,test the gene expression which about bone formation and bone resorption.Results By comparison,we found that blood biochemical indexes and PTH of patients of two groups have no statistical difference.25-hydroxyitamin D and PINP of osteoporosis group was significantly lower than the control group,and CXT and OC have not seen obvious difference.So,we could say that people of osteoporosis group are all the senile osteoporosis patients,and have lower abundance of lactobacillus and butylic acid bacteria.A high concentration of supernatant of lactobacillus acidophilus can stimulate proliferation,differentiation and maturity of osteoblasts,inhibit proliferation,differentiation and maturity of osteoblasts,enhance the gene expression of RunX-2 and ruduce the gene expression of RANK.However,LGG has not been found a similar effect.Conclusions Gut microbiota of osteoporosis patients are imbalance: abundance of lactobacillus and butylic acid bacteria are lower.The supernatant of lactobacillus acidophilus can stimulate proliferation,differentiation and maturity of osteoblasts,inhibit proliferation,differentiation and maturity of osteoblasts,enhance the gene expression of RunX-2 and ruduce the gene expression of RANK.