| Background:With the development of society and the improvement of people's health consciousness,the influence of high altitude climate on human reproductive health has been paid more and more attention.Existing studies have shown that high altitude and low pressure hypoxic environment will not only cause the damage of people's own reproductive function in the low pressure and hypoxic environment of Plateau,but also lead to the deformity of their offspring.Purpose:In order to solve the problem of damage to the reproductive health of highland workers and high altitude residents and to maintain the health of their offspring,this experiment is to study the mechanism of reproductive diseases in plateau low pressure and hypoxic environment.Methods:In this experiment,zebrafish was used as model animal to simulate the effect of high pressure and low oxygen on reproductive health of 6000 m in plateau.The male and female zebrafish in the control group were raised in the zebra fish circulating water system,and the male and female zebrafish in the experimental group used animal low-pressure oxygen chamber to simulate the high pressure and low oxygen treatment of the plateau 6000 m d.After the end of low oxygen and low pressure molding,he staining detected the morphological changes of gonadal tissue in male and female zebrafish,counted the gonadal index of male and female zebrafish,the spawning amount of females and the sperm survival rate of male zebrafish The difference gene of male and female zebra fish gonadal was obtained by high-throughput sequencing,and the function of zebra fish gonadal under low pressure hypoxic condition was obtained by using differential gene KEGG,and the signal pathway of zebra fish gonads under low pressure hypoxic condition was obtained by the enrichment of different genes.Specific primer screening and RT-q PCR verification of some differential genes were carried out to count the accuracy of high-throughput sequencing.Results: After simulated plateau 6000 m height low pressure hypoxic treatment,the female zebra Fish gonadal index decreased significantly,the egg cells in the ovary were loosely arranged,the number of oocytes decreased,the yolk granules of mature oocytes decreased,some egg cells were abnormal in morphology,the spawning volume of female zebrafish was significantly reduced,and the hatching rate of lateral F1 generation was significantly reduced.The deformity rate was significantly increased,and after the simulated plateau 6000 m height low pressure hypoxic treatment,the male zebrafish gonadal index decreased significantly,the seminal SAC structure was destroyed,the number of spermatocytes decreased,the sperm distribution was scattered,the sperm survival rate of male zebra fish decreased significantly,and the early apoptosis was significantly improved.The late apoptosis or mortality rate of male zebrafish sperm increased significantly,and in the lateral experiment of male zebrafish,the hatching rate of F1 was significantly decreased,and the deformity rate of F1 generation was significantly improved.There were 2,601 differential genes in female zebrafish,and go term,which was downgraded by go,had 40,covering the main functions of mitosis cell cycle regulation,negative regulation of apoptosis process,protein self-phosphorylation,protein kinase activity,protein phosphorylation,protein serine/threonine kinase activity,adenosine triphosphate binding,and cytoplasm There were 29 go term with go enrichment,and the main functions involved were extracellular clearance and lipid binding,and there were 3 KEGG term in KEGG enrichment,which were the biosynthesis and metabolism of oligosaccharides,the metabolism of auxiliary factors and vitamins,the treatment,folding,classification and degradation of genetic information.There were 6 KEGG term in KEGG,namely alpha-linolenic acid metabolism,linoleic acid metabolism,ether lipid metabolism,arachidonic acid metabolism,amino acid and nucleotide glucose metabolism,and peroxidase breeder activation receptor signaling pathway.There are 9,094 differential genes in male zebrafish,and go term has 31,and the main functions involved are the aggregation of dynamic protein family protein in mitochondrial division and the MHC? protein complex,and the main functions of go term are 7,involving cholesterol,fatty acids,phosphatidylcholine,Metabolism of very low density lipoprotein and very high density lipoprotein;KEGG increased KEGG Term 4,respectively,intestinal immune network to produce Ig A,cell adhesion molecules,phagocytic body,herpes simplex infection.Specific primer screening was carried out on some differential genes,41 pairs of specific primers were screened,the coincidence rate between female zebrafish high-throughput sequencing results and RT-q PCR results was 44%,and the coincidence rate between male zebra fish high-throughput sequencing and RT-q PCR results was 50%.Conclusion:This experiment proves that simulating 6000 m height low pressure hypoxic effect 14 d in Plateau can seriously damage zebra fish reproductive system and improve the birth defects of offspring.Plateau low-pressure hypoxia mainly affects the female zebra fish reproductive system through biological processes such as interfering with mitosis of ovarian cells,promoting ovarian cell apoptosis and extracellular clearance,and affects female zebra fish reproductive system mainly through signaling pathways related to the metabolism of various substances in cells.Plateau low-pressure hypoxia mainly affects the male zebra fish reproductive system by affecting the function of cell energy supply and immune response,and affects the male zebra fish reproductive system mainly by influencing the intestinal immune network to produce Ig A,cell adhesion molecule,phagocytic body,herpes simplex infection and other signaling pathways.The overall accuracy of high throughput sequencing of male and female zebra fish was 46.3%. |
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