Salidroside Activates Autophagy Of Corpus Cavernosum Smooth Muscle Cells Through AMPK/mTOR Signaling Pathway To Treat Erectile Dysfunction In Rats With Cavernous Nerve Injury

Objective:1.To explore the role of AMPK/mTOR signaling pathway in the regulation of autophagy and apoptosis of corpus cavernosum smooth muscle cells(CCSMCs);2.To explore whether salidroside activates autophagy of CCSMCs through AMPK/mTOR signaling pathway,so as to improve the erectile function of cavernous nerve(CN)injured erectile dysfunction(ED)rats.Methods:1.In vitro experiments:Primary CCSMCs were cultured in a hypoxic environment to establish hypoxia-induced oxidative stress apoptosis model.CCK-8 was used to screen the appropriate salidroside concentration and intervention.Western Blotting was used to detect the expression of key proteins in AMPK/mTOR signaling pathway.Western Blotting and immunofluorescence were used to observe the improvement of autophagy and apoptosis.2.The in vivo experiment:30 SD rats were randomly divided into control group,model group and treatment group,rhodiola rosea by surgical clip damage bilateral CN to establish the rat model with nerve injury sex ED,and practical rhodiola glucoside lavage intervention,control simulation procedure but not damage of nerve,control group and model group give amount of saline lavage.After four weeks of treatment,the erectile function of the rats was evaluated by penile sponge in vivo pressure test under electrical stimulation,and the penile sponge tissues were collected for Masson’s staining,Western Blotting,immunohistofluorescence and histochemistry to detect apoptosis,autophagy apoptosis,AMPK/mTOR signaling pathway related protein expression and smooth muscle relative content.Results:1.Primary CCSMCs were isolated.At low magnification,the cells were in vortex shape,and at high magnification,the cells were in long spindle shape,and the smooth muscle marker protein α-SMA was positive.CCSMCs treated with hypoxia and salidroside significantly increased p-AMPK/AMPK,decreased p-mTOR/mTOR,and increased autophagy related protein expression and decreased apoptosis related protein expression.These improvements disappeared or were reduced when Dorsomorphin,an AMPK inhibitor,was used to block AMPK/mTOR signaling.2.Erectile function of SD rats with bilateral CN clamp injury was significantly decreased,Masson’s staining results showed that the ratio of smooth muscle to collagen was decreased,Western Blotting results showed that p-AMPK/AMPK was significantly increased,and p-MTOR/mTOR was significantly decreased.Meanwhile,immunofluorescence and Western Blotting results showed that autophagy level in the cavernous tissue was increased,but the apoptosis level was still higher than that of the sham operation group.After salidroside treatment,erectile function of rats was significantly improved,the ratio of slippery muscle to collagen was increased,AMPK/mTOR signaling pathway was further activated,autophagy level was significantly increased and apoptosis level was reduced.Conclusion:1.AMPK/mTOR signaling pathway is an important pathway to regulate the level of autophagy in cavernosa tissue of RATS with CN injury;2.Salidroside can activate protective autophagy through AMPK/mTOR signaling pathway to treat CN damage ED.