The Protective Effect Of Cyanidin-3-O-Glucoside On Reproductive Injury In Mice Exposed To 3-Chloro-1,2-Propanediol

Objective:As a common food contaminant,3-monochloropropane-1,2-diol(3-MCPD)can target the testes and epididymis and cause a decrease in sperm quality,which is one of the reasons that causes male reproductive damage.The targeting effect of 3-MCPD on male reproductive system has aroused widespread concern.Cyanidin-3-O-glucoside(C3G),as an antioxidant anthocyanin that is widely ingested daily,shows good biological activity.This makes it a potential nutritional factor that antagonizes the reproductive damage caused by 3-MCPD.This project mainly explores the protective effects and mechanisms of C3 G on male reproductive injury induced by 3-MCPD in vivo by establishing a 3-MCPD-exposed reproductive injury model intervening with different dose of C3 G.Methods:6-week-old ICR male mouse were given 10 mg/kg·bw 3-MCPD solution daily by gavage to establish a reproductive injury model,and three groups of nutrition intervention groups were set up,given 25 mg/kg·bw,50 mg/kg·bw,and 100 mg/kg·bw respectively.Gavage intervention is with C3 G.And the experiment lasted 12 weeks.After the intervention,the serum,testis,epididymis,and sperm were separated and the relevant indicators were measured.(1)Histological evaluation of testis and epididymis by hematoxylin-eosin staining and scored spermatogenic epithelium is to characterize the protective effect of C3 G intervention on testicular tissue damage induced by 3-MCPD.(2)The computer-assisted semen analysis system was used to measure sperm quantity,deformity rate,viability,vitality and exercise-related parameters,which proved that C3 G intervention can protect the sperm quality caused by 3-MCPD.(3)The content of related hormones in serum were detected by enzyme-linked immunosorbent assay(ELISA)to explored the role of C3 G in regulating 3-MCPD-induced sex hormone secretion.(4)We use RNA-sequence technology to collect,compare,and analyze transcription sample data under 3-MCPD exposure and sample data after C3 G intervention to screen differentially expressed genes.By comparing with public databases GO and KEGG,unigene function annotation was performed,key expression processes were screened,and differentially expressed genes and pathways were analyzed.(5)Real-time quantitative PCR(Real-time quantitative PCR,q PCR)was used to analyze the expression of related differential genes to verify the regulatory effect of C3 G on gene expression caused by 3-MCPD.Results:(1)Compared with the 3-MCPD group,the 100 mg/kg·bw C3 G intervention group increased the weight gain of mice,improved the pathological damage of testes,and effectively improved the testicular seminiferous epithelium score.(2)C3G intervention increased sperm count,increased sperm motility and viability,improved sperm motility,reduced sperm deformity rate,and effectively improved sperm quality compared with the 3-MCPD group.(3)3-MCPD exposure can interfere with the secretion of sex hormones and cause abnormally high levels of FSH and T,decrease the level of LH and INH-B.C3 G can maintain normal hormone levels,allowing spermatogenesis to proceed normally.(4)3-MCPD exposure caused 69 genes to be significantly down-regulated,21 genes were significantly up-regulated,and C3 G antagonist 3-MCPD by up-regulating 38 genes and downregulating the expression of 22 genes.Through gene function analysis and pathway analysis,it was found that C3 G can effectively regulate cell cycle and immune regulation-related pathways,improve inflammatory effects,and C3 G intervention has obvious correlations with mitochondrial damage,sperm protease complex,endoplasmic reticulum.(5)3-MCPD down-regulates the expression of Fbxw15,affects HBO1 expression.C3 G can antagonize the Fbxw15 expression disorder caused by 3-MCPD,reduce the content of HBO1,regulate the normal activation of AR,provide a channel for T to enter the lumen,and stabilize the androgen environment required for spermatogenesis,regulate the testosterone in the testis to regulate the spermatogenesis,thereby improving sperm quality and testicular damage.Conclusion:3-MCPD exposure causes testicular damage and sperm quality decline in mice,while C3 G can maintain normal sex hormone synthesis and secretion,and improve testicular function and sperm quality.C3 G increases the level of testosterone and FSH to maintain the necessary hormonal environment during spermatogenesis,and provides protection for the reduction in sperm quantity and quality caused by hormonal levels.At the same time,C3 G affects HBO1 by intervening Fbxw15,thereby regulating the normal activation of AR,providing a channel for T to enter the lumen,and stabilizing the androgen environment required for spermatogenesis,providing a good environment for spermatogenesis and differentiation,to ensure sperm quality.This study can provide experimental basis for 3-MCPD-exposed people to make reasonable dietary guidance for more intake of anthocyanin foods,so as to reduce the decline of male sperm quality.