Se@Albumin Nanoparticles Ameliorate Intestinal Mucositis Caused By Cisplatin Through The Gut Microbiota Regulation

Objective: To investigate the specific molecular mechanism of Se@Albumin nanoparticles(Se@Albumin NPs)alleviating cisplatin(CDDP)-induced intestinal mucositis,and to provide a new strategy and scientific basis for the clinical prevention and treatment of chemotherapyinduced intestinal mucositis.Methods: 1.Through the self-assembly of denatured human serum albumin and selenite,a new form of Se@Albumin complex nanoparticles(Se@Albumin NPs)was developed,and its particle size,stability,and characterization were studied.2.The Se@Albumin NPs at a concentration of 0.1 mg/kg were administered to mice for one week,and the in vivo detection was performed by analyzing serum creatinine,urea nitrogen,alanine aminotransferase,aspartate aminotransferase,and histopathology of major organs toxicity.3.Divide the mice into four groups,namely: PBSsaline(PBS-saline)group,PBS-cisplatin(PBS-CDDP)group,nanoselenium-cisplatin(Se NPs-CDDP)group,Se@Albumin complex nanoparticles-cisplatin(Se@Albumin NPs-CDDP)group;gastric emptying test by mouse phenol red,mouse semisolid paste small intestine propulsion test,fluorescein isothiocyanate(FITC)-dextran method to detect gastric Intestinal motility and intestinal permeability confirmed the role of Se@Albumin NPs in CDDP-induced intestinal mucositis in mice.4.Enzyme-linked immunosorbent assay(ELISA)was used to detect IL-6,IL-10,TNF-α;kits were used to detect SOD,MDA,GSH,GPx-related indicators,and pathological HE staining was performed on intestinal tissue to explore Anti-inflammatory and antioxidant capacity of Se@Albumin NPs in the intestine of CDDP-induced mucositis mice.5.Collect the feces of four groups of mice in step 3 for 16 S rDNA sequence sequencing,analyze the distribution and abundance of intestinal microbiota in each group,and explore the effect of Se@Albumin NPs on the intestinal microbiota of mice with CDDP-induced mucositis.Influence 6.Obtain the feces of mice in each group in step5 as the donor,and use the antibiotic method to forge germ-free mice as the corresponding recipient group.Perform a fecal bacterial transplantation(FMT)experiment,transplant the feces to the recipient group of mice,and test again.Intestinal histopathological HE staining,ELISA assay to detect IL-6,IL-10,TNF-α,kit to detect SOD,MDA,GSH,GPx,and related indexes were also analyzed to confirm the Se@Albumin NPs.Can modulate intestinal microbiota to improve CDDP-induced intestinal mucositis in mice.Results: 1.Characterization of Se@Albumin NPs,transmission electron microscopy(TEM)uniform size is about 40 nm,zeta potential shows that Se@Albumin NPs have more negative charges(-37.3 m V),and the encapsulation efficiency of selenium is greater than 85 %,after 12 days of incubation in PBS at 25°C,there was no significant change in the size of Se@Albumin NPs.2.Histopathological HE staining,serum creatinine,urea nitrogen,alanine aminotransferase,aspartate aminotransferase detection of the main organs of the mice showed no obvious abnormality and tissue damage.3.Mouse phenol red gastric emptying experiment and mouse semi-solid paste small intestine propulsion experiment showed that Se@Albumin NPs could slow down the delayed gastric emptying caused by CDDP.Fluorescein isothiocyanate(FITC)-dextran experiment showed that Se@Albumin NPs could reduce intestinal permeability.4.ELISA experiments showed that Se@Albumin NPs inhibited the increase of proinflammatory factors IL-6 and TNF-α,and the decrease of antiinflammatory factor IL-10 in the intestinal tissue of mice after CDDP intervention;at the same time,it inhibited the increase of the peroxidation index MDA,and the anti-inflammatory cytokines.The levels of oxidative indexes GSH,GPx and SOD were decreased,and the tissue damage such as shortening of intestinal villi and shallowing of intestinal crypts caused by CDDP in intestinal mucositis were alleviated.5.16 S rDNA sequencing analysis showed that Se@Albumin NPs restored the intestinal microbiome imbalance caused by CDDP,mainly including the restoration of the abundance of anti-inflammatory bacteria(Bacteroidetes,Firmicutes)and the reduction of the abundance of pro-inflammatory bacteria(Escherichia).6.In the fecal bacteria transplantation experiment,the recipient mice in the fecal bacteria transplantation Se@Albumin NPs-CDDP group had lower weight loss,less damage to intestinal mucosa,and intestinal inflammation(IL-6,TNF-α,IL-10).Levels and peroxidation status(MDA,GSH,GPx,SOD)were alleviated.Conclusion: Se@Albumin nanoparticles ameliorate intestinal mucositis caused by cisplanin through the gut microbiota targeted regulation.