The Role Of Testosterone In The Development Of Polycystic Ovary Syndrome-associated Kidney Injury

BackgroundPolycystic ovary syndrome(PCOS)is one of the most common reproductive endocrine diseases in women of childbearing age,which manifests as menstrual disorders,polycystic ovaries changes,and hyperandrogenemia.The long-term complications of PCOS include obesity,type 2 diabetes and heart cardiovascular Diseases,etc.Chronic kidney disease(CKD)is the main cause of end-stage renal disease,which is closely related to the metabolic diseases.Proteinuria is known as one of the early clinical manifestations of CKD.It has been reported that more than 50%of PCOS patients have microalbuminuria associated with metabolic syndrome,suggesting a correlation between PCOS and renal injury.However,the specific manifestations and mechanisms of renal injury in PCOS remain unclear.As we all known,hyperandrogenemia can not only aggravate the clinical manifestations of PCOS,but also increase the risk of PCOS metabolic abnormalities.Previous studies have shown that serum androgen excess is closely related to the occurrence of tubular proteinuria.Hence,it is speculated that androgens may play an important role in PCOS-related kidney injury.Therefore,this study intends to explore the potential role and mechanism of androgen in PCOS-related kidney injury,expecting to provide theoretical support for the development of new measures for the prevention and treatment of PCOS-related kidney disease based on androgen.ObjectiveTo investigate the role and mechanism of serum testosterone in PCOS-related renal injury.Methods1.Relationship between serum testosterone and renal injury in patients with PCOS(1)Urine samples were collected from 55 PCOS patients and 69 healthy controls,which were analyzed by an automatic specific protein analyzer.The following indexes were detected:urinary Albumin to Creatinine Ratio(UACR),urinary k-light Chains(KapU),and urinary ?-light Chains(LamU),urinary?1-microglobulin(al-MU)and urinary ?2-microglobulin(?2-MU),which were normalized with urinary creatinine.Comparison between groups and correlation analysis were performed with the results(2)Follicular fluid and granulosa cells were collected from patients,five PCOS patients with hyperandrogenism,five PCOS patients with normal androgen and 5 healthy controls,respectively.The concentration of testosterone in the follicular fluid were tested using electrochemiluminescence.The mRNA expression of Caspase 3 and NLRP3 in granulosa cells were detected by qRT-PCR.2.Effects of follicular fluid from PCOS patients on human kidney proximal tubular epithelial cells fibrosisHuman kidney proximal tubular epithelial cells(HKC-8 cells)were stimulated with follicular fluid from PCOS patients with androgen excess.Western blot was used to detect the expression of fibronectin(FN)and ?-smooth muscle actin(a-SMA)in HKC-8 cells.Immunofluorescence was used to detect the expression of FN.The follicular fluid of normal women were used as controlsResults1.Relationship between serum testosterone and renal injury in patients with PCOS(1)The UACR in the PCOS group was significantly higher than that in the control group,suggesting that PCOS was related to the occurrence of kidney injury.(2)In the PCOS group,KapU,LamU,?1-MU,and ?2-MU were positively correlated with serum testosterone concentration,suggesting that serum testosterone was related to PCOS renal tubular injury.(3)The level of testosterone concentration in follicular fluid was significantly higher in the group of PCOS with hyperandrogenemia than controls and the group of PCOS with nonnal androgens.Similarly,the mRNA levels of Caspase3 and NLRP3 were highly expressed in granulosa cells,from the group of PCOS with hyperandrogenism,suggesting the level of testosterone were closely related to granulosa cells apoptosis and inflammation in PCOS2.Effects of follicular fluid from patients with PCOS on human renal tubular epithelial cell fibrosisThe result of western blot showed that the expression of FN and a-SMA were significantly up-regulated in HKC-8 cells treated with PCOS with hyperandrogenism.Likewise,results from immunofluorescence showed that the expression of FN was significantly up regulated.Results showed above suggested that high levels of testosterone could induce the occurrence of fibrosis in HKC-8 cellsConclusions1.PCOS is related to the occurrence of kidney injury.2.Serum testosterone plays an important role in PCOS-related kidney injury.3.Testosterone can mediate the occurrence of PCOS-related kidney injury by promoting fibrosis of renal tubular epithelial cells.