A Preliminary Study On The Improvement Of Oligozoospermia By Total Flavonoids From Semen Cuscutae Through GPRC6A

Objective: To explore the possible mechanism of TFSC to improve oligozoospermia through GPRC6 A.Methods: 1)Establish rat models of oligozoospermia in groups,and give TFSC and OCN interventions to record the changes in body weight,testicular index and femoral index of rats,count the number of sperm and activity rate;2)Testicular and femurs morphological changes were detected under light microscopy,and microstructural changes of testes were detected under electron microscopy;3)RT-qPCR method was used to detect the expression levels of gprc6 a and spermatogenesis-related genes ocn,creb and cyp11a1 in testicular tissue,and to detect the expression levels of gprc6 a and ocn in femoral tissue;4)Western Blot method was used to detect the expression of GPRC6 A and spermatogenesis-related proteins CREB,CYP11A1 and StAR in testis tissue;(5)ELISA method was used to detect the content of serum LH,FSH,T and OCN.Results: 1)The weight of rats in the M group was lower than that in the BC group,TFSC group and OCN group after modeling(P<0.05);the sperm count and activity rate of the M group rats were lower than those in the BC group(P<0.05),and the oligozoospermia model was successfully constructed;the sperm counts and activity rates of TFSC and OCN group were higher than those in M group(P<0.05);testicular index of rats in OCN group was lower than that in M group(P<0.05);there was no significant difference in femoral index between rats in each group(P>0.05);2)Under light microscope,the tissue structure of the testes of M group rats was damaged,the number of spermatogenic cells at each level was reduced,and the sperm cells were sparse in the lumen.The morphology of testes in TFSC and OCN groups was not much different from that in BC group.Complete structure,spermatogenic cells at all levels are tightly arranged,and the number of spermatids in the lumen is large.Electron microscopy shows that the number of spermatogenic cells at various levels in the testicular tissue of M group is reduced,the thickness is reduced,and the arrangement is disordered.The number of intracellular organelles was significantly reduced,and mitochondria were occasionally deformed.The testicular tissue structure of the BC group,TFSC group and OCN group is more complete,the seminiferous tubule basement membrane is continuous,and the number of spermatogenic cells at various levels is abundant.Under the light microscope,the morphology of the femur of each group of rats was not significantly changed,and the irregular inner ring bone plate and Haversian system were all visible;3)RT-qPCR results showed that in testis tissue,the expression level of gprc6 a in M group was lower than that in BC group(P<0.05);the expression level of ocn in OCN group was higher than that in BC group(P<0.05);the expression level of creb in TFSC group M group was high(P<0.05).In femoral tissues,there was no statistically significant difference in gprc6 a expression levels of rats in each group(P>0.05);the gene expression of femoral ocn in group M rats was lower than that in BC group,TFSC group and OCN group(P<0.05);4)Western Blot results showed that the protein expression of GPRC6 A,CREB,and CYP11A1 in testes of TFSC and BC groups was higher than that of group M(P<0.05);the expression of StAR protein in testes of TFSC group was higher than that of M,OCN group and BC group(P<0.05);StAR protein expression in testes of M group rats was lower than that of BC group(P<0.05);5)The levels of LH and FSH in serum of group M rats were higher than those of TFSC group,OCN group and BC group(P<0.05),and the levels of LH and FSH in serum of group BC rats were lower than those of TFSC group and OCN group(P<0.05);T content in serum of group M rats was higher than that of BC group(P<0.05);OCN content in serum of group M rats was lower than that of TFSC group,OCN group and BC group(P<0.05),OCN group The content of OCN in rat serum was higher than that in TFSC group and BC group(P<0.05).Conclusions: 1)Both TFSC and OCN have a certain therapeutic effect on oligozoospermia,and the effect of TFSC is more significant;2)TFSC may improve oligozoospermia by promoting the expression of gprc6 a in testis and GPRC6 A,CREB,CYP11A1 and StAR in testis.TFSC can further regulate the synthesis and secretion of T in Leydig cells of testis,affecting spermatogenic function and improving oligozoospermia.