| Objective:In this study,we established high-fat diet injury model of rats,the application of Danggui Buxue decoction to intervene at the same time,to investigate its protective effect on testicular injury caused by high fat diet,to further study the mechanism of Akt/Gsk3?signaling pathway in Danggui Buxue decoction to protect testicular injury in HFD rats,and to provide new ideas and theoretical basis for the prevention and treatment of high-fat induced testicular injury.Methods:Fifty SPF male Wistar rats(100-120g)were randomly divided into control group(Con group),high fat diet group(HFD group),Danggui Buxue decoction low-dose group(HFD+DBDL group),Danggui Buxue decoction high-dose group(HFD+DBDH group)and rosiglitazone control group(HFD+RGZ group),with 10rats in each group.The Con group was fed with standard normal diet,and the other groups were fed with high-fat diet without restriction of drinking water.The HFD+DBDL group and the HFD+DBDH group were given Danggui Buxue decoction solution 6 g·kg·d-1and 12 g·kg·d-1,respectively;the HFD+RGZ group was given rosiglitazone suspension 4 mg·kg·d-1,while the Con group and the HFD group were intragastrically administered with normal saline at the same volume.After continuous intragastrically administered for 12 weeks,complete testis tissues were taken from the rats,and sperm count and sperm motility were measured in the caudal epididymis.The changes of malondialdehyde(MDA)and superoxide dismutase(SOD)in testicular tissue will be detection by biochemical kit.HE was used to observe the microscopic staining of the left testis.TUNEL was used to observe the apoptotic staining of testis.Western Blot method was used to determine the apoptosis-related protein Bax Bcl-2 in testis;Pyroptosis associated proteins NALP3,Caspase-1,IL-1?,IL-18;Autophagy-related Beclin-1,LC3B and Akt/Gsk3?signaling pathway protein expression levels.Results:The total number and motility of sperm in HFD group were significantly lower than those in Con group,and the content of MDA in testis increased and the activity of SOD decreased,HE staining showed atrophy and degeneration of the convoluted seminiferous tubules and shedding of spermatogenic epithelium in high fat diet,The number of apoptotic bodies in the testis of rats in the HFD group was increased by TUNEL method,and the apoptotic rate increased significantly,which was statistically significant compared with that in the Con group(P<0.05).After low and high dose administration,Danggui Buxue decoction can improve sperm motility and total sperm count to normal level,reduce MDA content and increase SOD activity,protect testicular tissue structure,inhibit the formation of testicular apoptotic bodies,and reduce the apoptotic rate.Western blot results showed that:(1)The protein expressions of NALP3,Caspase-1,IL-1?and IL-18 in testis of rats in HFD group were significantly higher than those in Con group(P<0.05),Danggui Buxue decoction decreased the expression of pyroptosis protein in a dose-dependent manner(P<0.05).(2)The protein expressions of autophagy related proteins Beclin-1 and LC3B and the relative expression rates of apoptotic proteins Bax/Bcl-2 were significantly increased compared with Con group(P<0.05),and the protein expressions of Beclin-1,LC3B and Bax/Bcl-2 were down-regulated after Danggui Buxue decoction administration(P<0.05).(3)The protein expression ratio of p-Akt/Akt and p-Gsk3?/Gsk3?in HFD group was significantly decreased,and the difference was statistically significant compared with that in Con group(P<0.05).In a dose-dependent manner,the protein expression ratio of p-Akt/Akt and p-Gsk3?/Gsk3?was reversed by Danggui Buxue decoction(P<0.05).Conclusion:Danggui Buxue decoction has protective effects on testicular injury induced by high fat diet in rats.The mechanism may be related to the repair of testicular morphology and reproductive function,activation of Akt/Gsk3?signaling pathway,anti-oxidation,inhibition of testicular cell apoptosis,pyroptosis and regulation of autophagy level. |
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