| ObjectiveHypoxia may cause testicular congestion and edema,as well as reproductive function damage in male animals.It has been reported that Leptin is related to reproduction.This study explored the effect of exogenous leptin on reproductive function in male rats under hypoxia environment,and provided data for medical protection.MethodsExperiment 1:A hypobaric hypoxic chamber was used to simulate the environment at 5000m,and a hypoxic animal model was established.Male SD rats(12 weeks,401.0±12.5g)were randomly divided into 12 and 24 days hypoxic groups,and their opposite normoxia groups according to spermatogenesis cycle(12 days).Rats were anesthetized,left heart blood was collected,then rats were sacrificed.After that samples were collected.Weighed,testicular index and epididymis index were calculated.The total number,activity rate and abnormal rate of sperm were examined under microscope.HE staining detected morphology of testis and epididymis.The expression of hypoxiainducible factor-1α(HIF-1α)and nuclear factor-κB(NF-κB)in testicular tissues were detected by IHC staining.The apoptosis of testis cells was detected by TUNEL staining.Flow cytometry was used to detect the cell proportion of different ploidy in testis.Serum corticosterone(CORT),testosterone(T),free testosterone(FT)and the levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in testis were detected by ELISA.The levels of malondialdehyde(MDA),superoxide dismutase(SOD)and catalase(CAT)in testicular tissues were detected by chemical reaction colorimetry.The expression of NFκB in testicular tissues was detected by WB.Experiment 2:A hypobaric hypoxic chamber was used to simulate the environment at 5500m.Male SD rats(11 weeks,397.5±12.4g)were randomly divided into intraperitoneal injection of normal saline,5μg/kg Leptin,10μg/kg Leptin,30μg/kg Leptin hypoxia groups and the opposite normoxia groups.The rats were fed for 14 days,body weight was weighed daily and the reagents were injected intraperitoneally.Rats were anesthetized,left heart blood was collected,then rats were sacrificed.After that samples were collected.Weighed,testicular index and epididymis index were calculated.The total number of sperm was examined by microscopy.HE staining detected morphology of testis and epididymis.The expression of hypoxia-inducible factor-1α(HIF-1α)and nuclear factor-κB(NF-κB)in testicular tissues were detected by IHC staining.Flow cytometry was used to detect the cell proportion of different ploidy in testis.Immunofluorescence was used to detect the Leptin receptor(LEPR)level in sperm.The levels of CORT,T,FT,Leptin in serum,IL-6 and TNF-α in testis were detected by ELISA.The levels of MDA,SOD and CAT in testis were detected by chemical reaction colorimetry.The levels of NF-κB in testis were detected by WB.ResultsExperiment 1:Compared with normoxic group,in 12 and 24 days hypoxic groups,the body weight was significantly decreased(P<0.05),the sperm count and activity rate were significantly decreased(P<0.05),and the abnormal rate was significantly increased(P<0.05);HE staining showed thinning of spermatogenic epithelium,vacuolation of epithelial cells,enlargement of spermatogenic space,disordered arrangement of epididymal duct and reduced number of sperms in the duct.IHC staining showed increased expression of HIF-1α and NF-κB.TUNEL staining showed increased apoptosis of spermatogonia and spermatocyte.In 12 days hypoxia group,the serum T and FT levels were significantly decreased(P<0.05),MDA and SOD levels in testis were significantly increased(P<0.05),the levels of IL-6 and TNF-α in testis were significantly increased(P<0.05).In 24 days hypoxia group,the proportion of diploid to tetraploid cells in testis was significantly decreased(P<0.05).Compared with 12 days hypoxia group,the levels of IL-6 and TNF-α of testis in 24 days hypoxia group were significantly decreased(P<0.05).Experiment 2:Compared with normoxic group,in hypoxic groups,the body weight,food intake,testicular congestion and edema and increased capillaries were significantly decreased(P<0.05).HE staining showed that spermatogenic epithelium was disordered,spermatogenic tubule space was enlarged,epididymal duct arrangement was disordered and space was increased.IHC showed increased expression of HIF-1α and NF-κB in testis.Compared with normoxic normal saline group,MDA levels in testis in normoxic Leptin groups were significantly increased(P<0.05).Compared with hypoxic normal saline group,LEPR expression in sperm and IL-6 level in testis in 5μg/kg Leptin hypoxic group were significantly increased(P<0.05),and TNF-α level in testis in 30μg/kg hypoxic Leptin group was significantly decreased(P<0.05).ConclusionsSimulated hypoxic environment at 5000m and 5500m induced testis inflammatory injury,inflammatory response and oxidative stress were also increased.After exogenous Leptin was given to rats under normoxia,the level of MDA in testis was increased.The level of testis IL-6 increased in 5μg/kg Leptin group,while the level of testis TNF-αdecreased in 30 μg/kg Leptin group,after 14 days of continuous administration of exogenous Leptin in hypoxic environment at an altitude of 5500 m.Objective Through dynamically monitoring changes of cardiopulmonary function,sleep and mood states of the 37th Chinese Antarctic pre-selected expeditioners of Kunlun Station from Shanghai(4m)to Lhasa(3650m)and Yambajan(4300m),we obtained physiopsychological data at different altitudes,so as to screen hypoxia-susceptible expeditioners.Further,we would provide scientific data to select qualified expeditioners of Kunlun Station.Methods We used electrocardiograph,hemodynamic monitor,spirometer and actigraph to monitor cardiopulmonary function and sleep states of 27 healthy male expeditioners(32.9±6.1 years).A series of questionnaires were used to access acute mountain sickness(AMS)incidence,sleep and mood states.Results AMS incidences in Lhasa and Yambajan were 25.0%and 29.2%,respectively.Compared with Shanghai,in Lhasa and Yambajan,SpO2 significantly decreased,pulse rate and blood pressure significantly increased(P<0.001).HR and QTc significantly increased,QT,QRS,PP and RR significantly decreased(P<0.05).Cardiac contractility,cardiac work and cardiac pumping function significantly decreased,systemic vascular resistance(SVR)significantly increased(P<0.05).Thoracic fluid content(TFC)significantly decreased only in Yambajan(P<0.001).Airway patency significantly increased(P<0.05).Arousals during sleep significantly increased,sleep efficiency significantly decreased(P<0.01).Pittsburgh Sleep Quality Index(PSQI)score significantly increased(P<0.001).Vigor significantly decreased(P<0.05).Anxiety significantly increased only in Yambajan(P<0.01).Spearman correlation analysis revealed 61 physiological parameters had significant correlations with 10 psychological parameters,especially between PSQI score and fatigue(p=0.525,P<0.001),as well as between preejection period and tension(p=-0.429,P<0.001).Conclusion When expeditioners exposed to altitudes above 3650m,AMS incidence increased,cardiac function decreased,systemic vascular resistance increased,sleep disorders,and negative moods increased.Sleep and cardiovascular function were correlated with mood states. |
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