The Effect Of Type 2 Diabetes On Male Reproductive Function And The Regulation Mechanism Of Adiponectin

Objective:To observe the changes of adiponectin(APN)in male patients with type 2 diabetes mellitus(T2DM)and T2DM male rats before and after fasudil hydrochloride treatment.To explore the regulation mechanism of APN on male reproductive function and the effect of fasudil hydrochloride on testis.Methods:1.Clinical study:Forty male patients who underwent in vitro fertilization and embryo transfer assisted pregnancy in the department of reproduction medicine of the Second Hospital of Hebei Medical University were selected and divided into T2DM patients(N=20)and control group(N=20)according to fasting blood glucose.The general data,sex hormone and semen quality of the subjects were collected,later the levels of APN,interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in serum and seminal plasma were detected.The levels of APN,IL-6,TNF-α and their clinical indexes were compared.2.Animal study: Twenty-four 6-week-old male SD rats were divided into control group((N=6)and diabetic group.We prepared type 2 diabetic rats by intraperitoneal injection of streptozotocin(25mg/Kg).Eighteen rats were randomly divided into 3 groups: T2DM group(N=6);LF treatment group(N=6),HF treatment group(N=6).LF treatment group and HF treatment group were given Low dose of Fasudil Hydrochloride Injection(LF)10mg/kg and High dose of Fasudil Hydrochloride Injection(HF)30mg/ kg,respectively.Four months later,the experimental rats were euthanized and blood samples were collected.The testicular tissue on both sides was removed,weighed and the testicular index was calculated.The epididymis was removed and sperm suspension was prepared.Serum levels of testosterone and APN were determined by ELISA.The testicular tissue was observed by HE staining.The protein expression levels of APN,Adiponectin Receptor 1(AdipoR1)and Adiponectin Receptor 2(Adipo R2)in testis were analyzed by Western Blot and immunohistochemical methods.Relative data were compared between groups by one-way analysis of variance.Results:1.Clinical study :(1)Compared with the control group,the serum T level in T2DM group was significantly decreased(P<0.05);Sperm motility rate,sperm count and PR were significantly decreased(P<0.05).(2)Compared with the control group,serum and seminal adiponectin levels were decreased in T2DM group,and the difference in serum adiponectin levels was statistically significant(P<0.05).The levels of IL-6 and TNF-αin serum and seminal plasma were increased(P>0.05).(3)Serum TNF-α was negatively correlated with sperm concentration and sperm count(P>0.05).Serum IL-6was negatively correlated with total sperm count,total sperm viability and PR(P>0.05).2.Animal study:(1)Compared with the control group,T2DM group showed dark hair color,irritability and weight loss.(2)Compared with the control group,the body weight and sperm count of rats in T2DM group were significantly decreased(P<0.05).(3)Compared with the control group,the serum testosterone and adiponectin levels in T2DM group were significantly decreased,while the serum FBG level was significantly increased(P<0.05).Compared with T2DM group,the level of serum testosterone in HF group was significantly increased(P<0.05).(4)HE results showed that the convoluted seminiferous tubules of the testis in T2DM group were atrophied and spermatogenic cells were loosely arranged.Fasudil hydrochloride was improved after treatment.(5)Compared with the control group,the protein expression levels of APN,AdipoR1 and Adipo R2 in testis of T2DM group were significantly decreased(P<0.01).Compared with T2DM group,the protein expression levels of APN,AdipoR1 and Adipo R2 in testis of LF group and HF group were significantly increased(P<0.05).(6)The results of immunohistochemistry showed that the immunoreactivity of APN and Adipo-R2 was mainly in mesenchymal cells.The immunoreactivity of AdipoR1 was mainly in the seminiferous contractile duct.Compared with the normal group,the levels of APN,AdipoR1 and Adipo R2 in T2DM group were decreased(P<0.05).Conclusions:1.T2DM can affect male reproductive function,mainly manifested as abnormal sex hormones and semen quality.2.The mechanism of T2DM affecting male reproductive function may be related to tissue damage caused by decreased expression of APN and increased expression of inflammatory factors such as IL-6 and TNF-α.3.High-dose fasudil hydrochloride may reduce the protective effect of diabetes on testicular tissue damage by up-regulating the expression levels of APN,AdipoR1 and Adipo R2.