| Objective:1.To observe the effects of simulated low-pressure hypoxia environment on testicular organ index,sperm quality test,testicular tissue morphology,oxidative damage and serum sex hormone levels in Wistar malerats;2.To observe the effect of different concentrations of levocarnitine on the reproductive ability of Wistar male rats in the simulated low-pressure hypoxia environment of the plateau;Methods:1.Thirty-six 12-week male Wistar rats were randomly divided into four groups:group A blank group,group B model group,group C low dose group of 1-carnitine,and group D high dose group of 1-carnitine.Group A was raised outside the simulate high attitude and hypoxia chamber(local altitude:143 6m),whi le g roup B,group C and group D we re all raised in the simulated oxygen chamber at an altitude of 6,500m.Both the blank group of group A and the model group of group B received an abdominal injection of normal saline with the same volume as group C.Group C received 50mg/kg·d|-carnitine by intraperitoneal injection,while group D received 100mg/kg·d.After 28 days of intervention,rats in each group were weighed after intraperitoneal injection under anesthesia,blood was collected from abdominal aorta,and serum was centrifuged.2.Bilateral testicular tissues of rats were weighed to calculate the testicular tissue and organ index;Sperm suspensions were prepared from the tail of the epididymis on both sides,and sperm motility rate,sperm deformity rate,sperm count and sperm forward movement were observed under a microscope.3.Take each rat testicular tissue,left testicular tissue for wood grain-Eosin Staining(Hematoxylin-Eosin Staining,HE)after observing then Wistar rat testicular tissue morphology change,based on the right side of testicular tissue in the TEM(Transmission Electron Microscope and TEM)observed under Wistar rats SSCS and support cells in testicular tissue three-dimensional ultra structure.The remaining right testicular tissue was taken to prepare homogenized serum,and the lipid peroxide metabolite malondialdehyde(MDA),Superoxide Dismutase(SOD)and Glutathione Peroxidase(GSH-PX)were detected by using the kit.4.Analysis of using Elisa kits in serum Testosterone(Testos terone,T),Luteinizing Hormone,Luteinizing Hormones,LH),follicle-stimulating Hormone,Follicle Stimulating Hormone,FSH)andsex hormone levels.Results:1.I n the simulated low-pressure and low-oxygen environment at the plateau,the org an index of the test is of male Wi star rats decreased in group B,group C and group D compared with group A(P<0.01).Compared with group B,organ indexes of group C and group D were increased(P<0.01).Compared with group C,organ index increased in group D(P<0.05).2.Compared with group A,the sperm motility rate and forw ard movement of group B,group C and group D decreased,and the sperm malformation rate increased(P<0.01),while the sper m count of group B decreased(P<0.01).Group C and group D compared with group B,live sperm rate and sperm count were significantly increased,sperm deformity rate significantly decreased(P<0.01);Group C(P<0.05)and group D(P<0.01)forward movement were obviously increased;Compared with g roup C,the sperm deformity rate group D was increased in group D(P<0.05).3.HE staining was used to observe the group A.The sperm at ogenic microtubule basement membrane of rat testicular tissue was complete,indicating the whole process of spermatogenesis,with A large number of spermatozoa in the lumen,no obvious damage to spermatozoa,orderly cell arrangement and regular cell polarity,and Johnsen score(9.40±0.38).Compared with group A,the testicular tissue of model group B rats showed atrophy,thinning,folding and stratification of the spermatogenic microtubule basement membrane,with obvious vacuoles,low sperm count,and A few early sperm at ogenic cells with disordered cell arrangement,decreased layers and loss of cell polarity.Johnsen score of model group B was(5.83±0.64).However,the rat testicular tissue in group C and group D showed slight folds,atrophy and stratification in the spermatogenic microtubule basement membrane,to increase the number of sperm,sperm damage is not apparent,lack of born mature sperm cells,cells is arranged with A little disorder,sparse,layer number 3-5,cell polarity can be seen,but the pathological change is still better than A blank group,group CI-carnitine Johnsen score low dose group(8.39+/-0.49),group DI-carnitine Johnsen score high dose group(8.29+/-0.64).Spermatogonium and supporting cells in rat testicular tissues of each group were observed by TEM.Normal spermatogonium and supporting cells we reobserved in the blank group A with clear structure,intact basal membrane,A small number of lipid droplets,no swelling of mitochondria,and no expansion of endoplasmic reticulum.In the model group B,a large number of vacuoles were formed in the cytoplasm,organelles were oedema and cavitation was obvious,mitochondria were ruptured and swollen,and lipids were abnormal.The nuclear membrane of interstitial cells was ruptured,mitochondrial edema was observed,local cytoplasm was absent,cavitation was observed,nuclear heterochromatin was increased,and endoplasmic reticulum was expanded.A few vacuoles were observed around the spermatogonium membrane,and the cell membrane was damaged obviously.The primary spermatocyte endoplasmic reticulum was slightly swollen,the cell membrane was fuzzy,and the nuclear heterochromatin was increased.Large vacuoles form around secondary spermatocytes.In group C,the supporting cells in the low-dose levocarnitine group were slightly detached from the seminiferous epithelium,the structure of cell membrane and nuclear membrane was still clear,and the mitochondria were slightly swollen.The endoplasmic reticulum of mesenchymal cells expanded slightly,the nuclear membrane of spermatogonium was clear,and the mitochondria were normal.In thehigh-dose levocarnitine group,the supporting cell structure was still clear,a few vacuoles were observed in the cytoplasm,and mitochondria were slightly swollen.The structure of mesenchymal cells,spermatocytes and primary spermatocytes were clear,the basal membrane was intact,there was no swelling of mitochondria and no swelling of the endoplasmic reticulum.4.MDA:compared with the blank group of group A,the modelgroup of group B,the low-dose group of group C and the high-dosegroup of group D all increased(P<0.01);co mpared w it h the model group of group B,the low-dose group of group C and the high-doseg roup of group D all decreased(P<0.01).SOD:compared with the blank group of group A,the model group of group B(P<0.01),the low-dose group of group C(P<0.01)and the high-dose group of group D(P<0.05)all decreased,while the low-dose group of group C and high-dose group of group D increased(P<0.01)compared with the model group of group B.GSH-PX:compared with the blank group of group A,the model group of group B,the low dose group of levocarnitine group C and the high dose group of levocarnitine group D all decreased(P<0.01),while the low dose group of levocarnitine group C and the high dose group of levocarnitine group D increased(P<0.01).5.Testosterone T:Compared with the blank group of group A,the model group of group B,Group C and the high-dose levocarnitine group of group D all decreased(P<0.01).LH:there was no difference between the groups.FSH decreased in model group B(P<0.05),low-dose group C(P<0.05)and high-dose group D(P<0.01).Conclusions:1.Simulated low-pressure and low-oxygen environment at hig h altitude can reduce the testicular organ index of male Wistar ra ts;Reduce sperm motility,forward motion,sperm count,and increa se the malformation rate;Testicular tissue morphology damage,oxid ative damage and serum sex hormone level change;2.L-carnitine has a certain protective effect on the reproduc tive capacity of male Wistar rats in the simulated low-pressure and low-oxygen environment at high altitude,100mg/kg,and the effe ct isbetter at 1/day. |
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