GR-Mediated Intrauterine Programming Regulation CX43 to Study the Mechanism of Induced Primary Spermatogenic Dysfunction in Offspring with Deficiency of Renal Es-sence of Female Rat
Objective To clarify the mechanism of primary testicular spermatogenic dysfunction in offspring induced by deficiency of renal essence in Maternal generation.Methods 24 pregnant rats were prepared and randomly divided into blank group,model group,and kidney tonifying group;In addition to the blank group,the remaining groups used chronic stress to replicate the mater-nal model of kidney essence deficiency during pregnancy.Enzyme-linked immunosorbent assay(ELISA)was used to detect ma-ternal serum thyroxine(T4),glucocorticoid(GC),and fetal number to evaluate the maternal model.Starting from the Oth day of pregnancy,the kidney tonifying group was given Zuogui Pill(0.39 mg/Kg)by gavage to fill the kidney essence;At the 20th day of pregnancy,compare the general situation and weight changes of female rat.Take male fetal rat,and detect their serum T4,GC,and FSH levels by ELISA.Evaluate the development of testicular germ cells by hematoxylin eosin staining.Immunofluores-cence double staining is used to detect the expression of Cx43 and Sox9 proteins in the testicular tissue of fetal rat.Real-time fluorescence quantitative polymerase chain reaction is used to detect the expression of ERK and Cx43 genes in the testicular tissue of fetal rat.Protein immunoblotting is used to detect GR Expression of ERK1/2,Cx43,and FSHR proteins in fetal rat testis.Re-sults Compared with the blank group,the maternal weight in the model group decreased,the number of fetal births decreased,the serum T4 content decreased,and the serum GC content increased(P＜0.01);Compared with the model group,the mothers in the kidney tonifying group had increased postpartum weight,increased fetal number,increased serum T4 content,and de-creased GC(P＜0.01).Compared with the male fetal rats in the blank group,the serum T4 content in the model group de-creased,the GC and FSH content increased,the testicular body mass ratio decreased,the number of sertoli cells,spermatogonia,and the number of seminiferous tubules decreased,the Sox9 and Cx43 proteins in the testicles decreased,the expression of ERK and Cx43 mRNA decreased,the expression of FSHR increased(P＜0.01 or P＜0.05),and the GR protein in placental tissue increased(P＜0.05);Compared with the model group,the fetal rats in the kidney tonifying group had an increase in serum T4 content,a decrease in GC and FSH content,an increase in testicular body mass ratio,an increase in the number of sertoli cells,spermatogonia,and the number of seminiferous tubules,an increase in testicular ERK and Cx43 mRNA expression,an increase in Sox9 and Cx43 proteins,a decrease in FSHR expression(P＜0.01 or P＜0.05),and a decrease in placental GR protein(P＜0.05).Conclusion Kidney essence deficiency during pregnancy in female rats is one of the predisposing factors for primary tes-ticular spermatogenesis dysfunction,and its mechanism may be related to GR/ERK/CX43 intrauterine programming to regulate the number of sertoli cells.
Maternal generationDeficiency of kidney essenceTesticular developmentIntrauterine programmingGlucocorticoidsGap junction protein 43
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