Research On The Mechanism Of Kidney Tonifying Herbs On Apoptosis Of Sertoli Cells Induced By Glycosides Of Tripterygium Wilfordii Hook F

Objective Based on Akt/NF-κB signaling pathway,to study the intervention mechanism of kidney tonifying compound Chinese medicine on Glycosides of Tripterygium wilfordii Hook F induced apoptosis of Sertoli cells,so as to provide scientific basis for the correct application of Glycosides of Tripterygium wilfordii Hook F in clinic.Methods 1.Preparation of serum containing drugs: 36 male SD rats of SPF grade were randomly divided into three groups according to their body weight: serum free group,Glycosides of Tripterygium wilfordii Hook F group and kidney tonifying Chinese medicine group.Carboxymethylcellulose sodium,Glycosides of Tripterygium wilfordii Hook F and kidney tonifying herbs were infused into the stomach respectively,and blood was taken for standby after7 days of continuous infusion.2.Intervention of Sertoli cells with serum containing drugs: cell proliferation rate was measured by MTT technology,cell cycle was measured by flow cytometry,caspase-9,caspase-3 protein and p-Akt,p65 protein were detected by Western blot.Results1.MTT resultsCompared with the blank control group,the cell proliferation rate of Tripterygium wilfordii Hook F group and lipopolysaccharide group decreased(P<0.05).2.The results of cell cycle detection by flow cytometryCompared with the blank control group,the proportion of G1 phase cells in Tripterygium wilfordii Hook F group and lipopolysaccharide group increased significantly,while the proportion of S phase cells decreased significantly(P<0.05).3.Expression of Caspase-9 and Caspase-3 in Sertoli cellsCompared with the blank control group,the expression of caspase-9 and Caspase-3 in Tripterygium wilfordii glycoside group and lipopolysaccharide group increased(P<0.05).4.Expression of p-Akt and p65 in Sertoli cellsCompared with the blank control group,the expression of p-Akt and p65 protein in Tripterygium wilfordii Hook F group and lipopolysaccharide group increased(P<0.05).Conclusion1.Tripterygium wilfordii Hook F may activate Caspase-3 to induce waterfall effect by up regulating the expression of Caspase-9,and activate the apoptosis pathway through Akt/NF-κB signal pathway,leading to the increase of testicular Sertoli cells apoptosis and reproductive damage.2.Another way of action of Tripterygium wilfordii Hook F on reproductive damage may be to block the division of Sertoli cells in S phase by affecting G1 phase and S phase of cells.3.It is speculated that the mechanism of kidney tonifying herbs against reproductive damage is that kidney tonifying herbs can antagonize the damage of Tripterygium wilfordii Hook F on Sertoli cells.Kidney tonifying herbs can restore DNA synthesis and inhibit the expression of apoptotic protein in injured cells,and play the role of promoting injury repair through Akt/NF-κB pathway.