Study On The Mechanism Of Combined Zinc And Selenium To Improve The Reproductive Damage Of Tripterygium Glycosides In Male Rats

Tripterygium glycosides are commonly used in clinical treatment of a variety of immune-related diseases.Long-term use can easily cause male reproductive damage.The mechanism mainly involves reproductive endocrine abnormalities,oxidative damage,apoptosis,and spermatogenesis disorders.Zinc and selenium are natural antioxidants with antioxidant and anti-inflammatory biological effects.They have synergistic effects in the male reproductive system,and the combined treatment effect is better than the single preparation.In this study,a male rat reproductive injury model induced by tripterygium glycosides was established,and combined with zinc and selenium treatment was given to explore the effect and mechanism of zinc and selenium combined intervention on improving reproductive injury induced by tripterygium glycosides in male rats.Methods:1.21 male rats were randomly divided into three groups : control group,model group(30 mg/kg/d tripterygium glycosides),zinc and selenium group(zinc(10 mg/kg/d)and selenium(0.1 mg/kg/d)),and control group was given the same amount of normal saline.2.To detect the total number of sperm and sperm motility in rats;serum testosterone levels were detected by ELISA;the pathological changes of testicular tissue were observed by HE staining.3.The m RNA and protein expression of zinc transporters Zn T10,ZIP8 and ZIP14,metal transcription factor(MTF1)and metallothionein(MT)in rat testis were detected by real-time fluorescence quantitative PCR(q PCR)and Western blotting(WB).4.The contents of zinc and selenium in serum and testicular tissue of rats were detected by inductively coupled plasma mass spectrometry;the activities of alcohol dehydrogenase(ADH),alkaline phosphatase(ALP)and lactic dehydrogenase(LDH)in testicular tissue were detected by colorimetry.5.Detection of antioxidant enzyme activity in testicular tissue by colorimetry;WB was used to detect the expression of PI3K/AKT/Nrf2/HO-1 pathway-related proteins in testicular tissue.Results:1.The rats in tripterygium glycosides exhibited a significant decrease in body weight and testicular weight(P<0.05);conversely,those in the zinc and selenium group displayed a significant increase(P<0.05).2.Rats exposed to tripterygium glycosides experienced a marked decrease in total sperm count,sperm motility and serum testosterone content(P<0.05),whereas rats exposed to zinc and selenium showed a significant rise in total sperm count,sperm vitality and serum testosterone content(P<0.05).3.HE staining revealed the control group’s testis structure to be normal,with seminiferous tubules arranged in close and neat proximity,interstitial cells visible,and spermatogenic cells arranged in a regular and orderly fashion.The lumen was full of mature sperm,while the model group’s seminiferous tubules were disordered,with a greater gap between them.The zinc-selenium group displayed a neat arrangement of testicular seminiferous tubules,a full lumen of these tubules,a significant increase in endogenous sperm cells,and a large number of mature sperm.In contrast,the wall of the tube had shrunk,its morphology was irregular,interstitial cells had dispersed,vacuolated seminiferous tubules,and various spermatogenic cells had significantly reduced or vanished.4.The m RNA expression of Zn T10 was significantly increased(P<0.05)and m RNA and protein expression of ZIP8,ZIP14 and MTF1 were significantly decreased(P<0.05)in the testis tissue of rats with tripterygium glycosides;the expression of Zn T10 was significantly decreased(P<0.05)and the expression of ZIP8,ZIP14 and MTF1 were significantly increased(P<0.05)in the zinc and selenium group.5.In rats exposed to tripterygium glycosides,serum and testicular tissues exhibited a decrease in zinc and selenium levels,while ADH,LDH and ALP activities were significantly diminished(P<0.05).Moreover,GPx1,Sepp m RNA and protein expression were significantly decreased(P<0.05);however,the zinc and selenium group showed a significant increase in the aforementioned indices.6.The testes of rats with tripterygium glycosides exhibited a significant decrease in SOD activity(P<0.05)and an increase in MDA activity(P<0.05).Conversely,the zinc and selenium group displayed a significant increase in SOD activity and a significant decrease in MDA activity(P<0.05).7.Rat testicular tissues exhibited a decrease in protein expression of PI3 K,p-AKT,Nrf2 and HO-1 when exposed to Tripterygium glycosides(P<0.05);however,the zinc and selenium group showed an increase in these indicators(P<0.05).Conclusion:1.Tripterygium glycosides caused testicular structural damage,decreased total sperm count and sperm motility in male rats,resulting in reproductive damage.Zinc and selenium intervention significantly improved reproductive damage in rats.2.The content of zinc and selenium in serum and testicular tissue of rats in tripterygium glycosides group decreased,the expression of ZIP8 and ZIP14 in testicular tissue decreased,the expression of Zn T10 increased,and the expression of MTF1 decreased,resulting in the imbalance of zinc homeostasis and selenium homeostasis.After combined treatment with zinc and selenium,the zinc homeostasis and selenium homeostasis of rat testicular tissue were significantly restored.3.The activity of ADH,ALP,LDH and the expression of selenoprotein Gpx1 and Sepp in the testis of rats in the tripterygium glycosides group were significantly decreased.After zinc and selenium intervention,the activity of zinc-containing enzyme ADH,ALP,LDH and the expression of selenoprotein Gpx1 and Sepp in the testis of rats were significantly increased.4.Zinc and selenium enhance the activity of antioxidant enzymes by activating the PI3K/AKT/Nrf2/HO-1 pathway,improve the oxidative damage induced by tripterygium glycosides,and then improve reproductive damage.