Polysaccharide Isolated From Enteromorpha Ameliorate Intestinal Microbiota And Intestinal Barrier In Mice

The application chemotherapeutic drugs such as Cisplatin(DDP)is limited due to adverse effects such as gastrointestinal toxicity and nephrotoxicity.Preliminary evidence has suggested that cisplatin injures small intestine.Intestinal barrier and intestinal microbiome are inextricably related,intestinal physiological bacteria constitute the intestinal biological barrier.Our previous study found that cisplatin leads to intestinal homeostaisis imbalance.To find the link between intestinal injury and intestinal microbiota imbalance caused by cisplatin can provide a theoretical basis for coping with cisplatin-induced intestinal side effects in clinical practice.Enteromorpha(EP)is a kind of green algae often consumed by residents living in the eastern coast of China.According to our previous study,polysaccharide from EP(PEP)rebalance the intestinal microbiota in constipated mice.Thus,we speculate that PEP can optimize the intestinal microbiota and help resisting further damage caused by DDP.Based on the background and pre-experimental basis,we treated mice with cisplatin for differen durations to explore the relationship between cisplatin-induced intestinal barrier damage and intestinal microbiota disorder.And pretreatment of mice with PEP to demonstrate that PEP can reduce the intestinal microbiota disorder and intestinal barrier damage.Objective Firstly,to find out the order between the intestinal damage caused by cisplatin and the disorder of intestinal microbiota.Secondly,intervene the mice with PEP,to demonstrate that PEP can interfere with the intestinal damage caused by cisplatin.Methods 1.The polysaccharide was extracted by water extraction and the polysaccharide was concentrated using the polysaccharide precipitated in 60%-80% alcohol.2.Animal experiments,BALb/ c mice female were divided into three groups,No treated group(No),cisplatin group(DDP),cisplatin and PEP group(DDP + PEP).The mice in DDP + PEP group were interfered with PEP,after one week,mice in DDP and DDP + PEP groups gained a single intraperitoneal injection with 14 mg / kg dose of cisplatin.Mice in DDP group were sacrificed at different time point: 0 h,12 h,36 h and 72 h,and the faeces of mice sacrificed at 72 h were collected.Then after 72 h,all mice were sacrificed and fecal,blood,small intestine,colonization in small intestine wall were collected.3.The apoptosis of intestinal epithelial cells in each group was detected by Td T-mediated d UTP end labeling(TUNEL)assay.The bacterial 16 s DNA sequencing was performed to study the differences of intestinal microbiota in the each group.H & E staining was used to determine clear intestinal damage caused by cisplatin.Immunohistochemistry staining was performed to indicate the damages in small intestine mechanical barrier,chemical barrier,immune barrier in situ.Finally,PCR-DGGE and 16 s DNA sequencing were used to detect small intestinal wall primary colonization bacteria(biological barrier).Results 1.The disorder in intestinal microbiota in ciaplatin injective mice was detected after 12 h after injection,and histopathology was detected after 36 h.2.The intestinal microbiota of mice in DDP group were disordered,the content of proteobacteria increased;the intestinal epithelial cell apoptosis was obvious,the mechanical barrier of intestine was damaged,the number of mucous protein expressing cells in the chemical barrier was significantly decreased,and Ig A positive cells substantial reduced.Escherichia-Shigella was increased in the small intestine in DDP group.3.Intestinal microbiomes tended to be normal under the prevention of PEP;the small intestinal barrier remained damaged in different degrees,but the damages were significantly reduced.In small intestine,Lactobacillus increased and pathogenic bacteria decreased significantly compared with DDP group.Conclusion 1.Cisplatin not only causes the destruction of the small intestine barrier,but also leads imbalance of intestinal microbiota;2.PEP intervention can reduce the disorder of intestinal microbiota caused by cisplatin;PEP intervention can reduce the cisplatin-induced intestinal damage.This study demonstrated that PEP plays a positive role in reducing the damage caused by cisplatin,which provided a theoretical basis for clinical practice to improve cancer patients’ quality of life.