Background:Bladder outlet obstruction(BOO)is a prevalent urinary system disease main caused by benign prostatic hyperplasia(BPH)in males.Traditional Chinese medicine(TCM)Danzhi qing’e decoction(DZQE)has the functions...Background:Bladder outlet obstruction(BOO)is a prevalent urinary system disease main caused by benign prostatic hyperplasia(BPH)in males.Traditional Chinese medicine(TCM)Danzhi qing’e decoction(DZQE)has the functions of strengthening“Yang Qi”(one of the fundamental concept in TCM,representing the active,warming,and energizing force within the body.It governs physiological functions,maintains body temperature,and promotes vitality.Balanced“Yang Qi”supports immunity and metabolism,while deficiency may lead to fatigue,cold intolerance,or weakened resilience),promoting blood circulation,and removing blood stasis,and nourishing yin based on the TCM theory.Previous studies have found that it significantly improved BPH and regulate urinary function in estrogen and androgen-induced rats.However,it is unclear whether DZQE has an inhibitory effect on BOO rats.Methods:Male Wistar rats underwent retropubic partial bladder neck ligation to induce BOO.DZQE extract(2.7/5.4 g/kg)was administered orally for 35 days.Anesthetized rats underwent cystometry to assess BOO and treatment effects on urinary parameters.Bladder histopathology,fibrosis,and PCNA expression were evaluated via HE,Masson’s,and IHC staining.Western blot quantified bladder tissue levels of choline acetyltransferase(ChAT),rho-associated protein kinase 1(ROCK1),myosin light chain kinase(MLCK),myosin light chain 2(MLC-2),extracellular signal-regulated kinase(ERK),proliferating cell nuclear antigen(PCNA),B-cell lymphoma-2(Bcl-2),and Bcl-2 related X protein(Bax).Primary bladder smooth muscle cells were cultured,exposed to hydrostatic pressure(HP,3 h/24 h)using a custom apparatus to mimic BOO,and treated with DZQE components for Western blot analysis of ROCK1,MLCK,ERK,PCNA,and Bax.Results:The bladder hyperplasia,bladder index(BI)increased,and histopathological alteration were easily observed in BOO group,which were significantly inhibited by DZQE administration.DZQE also significantly inhibited the up-regulation of maximum voiding pressure(MVP)and down-regulation of residual urine volume(RV)observed in BOO rats.The expressions of ChAT and MLCK and the activation of ERK were much increased,while the expressions of ROCK1,MLC-2 and Bax were obviously decreased in BOO rats,all of which were then significantly inhibited by DZQE.In rat bladder smooth muscle cells(RBSMC),3 h or 24 h duration of HP successfully simulated the BOO compensation and decompensation respectively in vitro.DZQE or its active components reduced the abnormal gene expressions in HP stimulated RBSMC.Conclusion:DZQE improves the urinary function in BOO rats mainly through the activation of ERK.Bakuchiol,salvianolic acid A,kaempferol,and tanshinoneⅡA are possibly the important active components of its therapeutic effects.展开更多
基金supported by The National Key Research and Development Project of China(No.2023YFC3504302)the National Natural Science Foundation of China(General Program No.82074105).
文摘Background:Bladder outlet obstruction(BOO)is a prevalent urinary system disease main caused by benign prostatic hyperplasia(BPH)in males.Traditional Chinese medicine(TCM)Danzhi qing’e decoction(DZQE)has the functions of strengthening“Yang Qi”(one of the fundamental concept in TCM,representing the active,warming,and energizing force within the body.It governs physiological functions,maintains body temperature,and promotes vitality.Balanced“Yang Qi”supports immunity and metabolism,while deficiency may lead to fatigue,cold intolerance,or weakened resilience),promoting blood circulation,and removing blood stasis,and nourishing yin based on the TCM theory.Previous studies have found that it significantly improved BPH and regulate urinary function in estrogen and androgen-induced rats.However,it is unclear whether DZQE has an inhibitory effect on BOO rats.Methods:Male Wistar rats underwent retropubic partial bladder neck ligation to induce BOO.DZQE extract(2.7/5.4 g/kg)was administered orally for 35 days.Anesthetized rats underwent cystometry to assess BOO and treatment effects on urinary parameters.Bladder histopathology,fibrosis,and PCNA expression were evaluated via HE,Masson’s,and IHC staining.Western blot quantified bladder tissue levels of choline acetyltransferase(ChAT),rho-associated protein kinase 1(ROCK1),myosin light chain kinase(MLCK),myosin light chain 2(MLC-2),extracellular signal-regulated kinase(ERK),proliferating cell nuclear antigen(PCNA),B-cell lymphoma-2(Bcl-2),and Bcl-2 related X protein(Bax).Primary bladder smooth muscle cells were cultured,exposed to hydrostatic pressure(HP,3 h/24 h)using a custom apparatus to mimic BOO,and treated with DZQE components for Western blot analysis of ROCK1,MLCK,ERK,PCNA,and Bax.Results:The bladder hyperplasia,bladder index(BI)increased,and histopathological alteration were easily observed in BOO group,which were significantly inhibited by DZQE administration.DZQE also significantly inhibited the up-regulation of maximum voiding pressure(MVP)and down-regulation of residual urine volume(RV)observed in BOO rats.The expressions of ChAT and MLCK and the activation of ERK were much increased,while the expressions of ROCK1,MLC-2 and Bax were obviously decreased in BOO rats,all of which were then significantly inhibited by DZQE.In rat bladder smooth muscle cells(RBSMC),3 h or 24 h duration of HP successfully simulated the BOO compensation and decompensation respectively in vitro.DZQE or its active components reduced the abnormal gene expressions in HP stimulated RBSMC.Conclusion:DZQE improves the urinary function in BOO rats mainly through the activation of ERK.Bakuchiol,salvianolic acid A,kaempferol,and tanshinoneⅡA are possibly the important active components of its therapeutic effects.
