Explore The Etiology,pathogenesis And Treatment Of Erectile Function Induced By The DEHP Based On The Theory Of Turbidity Toxin

The first part The discussions about the male reproductive toxicity of DEHP based on the theory of turbidity toxinObjective Discuss about the male reproductive toxicity of DEHP based on the theory of turbidity toxinMethods Combined with the progress of male reproductive toxicity of DEHP,we discuss the DEHP’s concept,the pathogenesis and treatment based on the theory of turbidity toxin,which is polluted air in the environment,is stored for poison intangibly,flows downward,and tends to damage kidney.Results We defines the DEHP as polluted air in the environment,which enters the body due to Qi activity by eating,breathing and so on.DEHP stays in the Qi and blood in the body,by which,DEHP flows throughout the body and zang-fu organs;DEHP is weightier and viscous,so that causes qi and blood stasis.What is more,DEHP could be stored in the lower-Jiao to damage the testis,including Leydig cells,Sertoli cells,sperm cells.Which eventually leads to that kidney Yang is not fair,kidney Yin is not solid,kidney essence is not filled,qi and blood is deficiency;In brief,the disease occurs in the qi and blood,and involves the spleen,liver and kidney three Zang organs.Turbidity toxin’s characters goes through the whole course of male reproductive toxicity of DEHP development;In the treatment of turbid poison DEHP,strengthening and restoring effects should be taken along with the reducing the toxicity.meantime,inspect syndrome comprehensive treatment based on syndrome differentiation.Conclusion The theory of "Turbid poison" in TCM well explains the characteristics of male reproductive toxicity of DEHP and guided treatment.The second part Explore the mechanism of ED induced by the DEHP on ERβ-eNOS signaling pathway and apoptosis mechanismObjective Observe the influence of DEHP on adolescent rats’ erectile function and discussed the mechanism.Methods 24 rats are randomly divided into 4 groups,each group had 6,respectively named corn oil group(the control group),low middle,high dose group of DEHP,respectively filled with corn oil 1 ml/d,DEHP10mg/kg/d,100mg/kg/d,1000mg/kg/d,for 14 consecutive days.24 hours after the last the treatment,APO experiments are taken,to observe the incubation period and the number of erection.The level of serum E2 and T is analysised by ELISA.The penile tissues are section,and then stained by HE,Masson,TUNEL and IHC,to analysis the expression of ER beta,eNOS and caveolin-1.The expressions of eNOS mRNA and caveolin 1 mRNA are analysised by RT-PCR,and the expressions of Bcl-2 and bax protein are analysised by western blotting.Results The general situation and the weight of each group before and after the experiment have no significant differences.Compared with controls,the erectile incubation period extends in the middle and high dose group(P<0.05);The rats in the middle and high dose groups erectile less(P<0.05);E2 concentration reduces in the low,middle and high dose group(P<0.05);But T concentration reduces in the middle and high dose group(P<0.05).By HE staining,the CCSM cells are long spindle,and blood sinus,endothelial cells and capillaries cavity’s structure are clear in the control group;The CCSM cells decreases,which is irregular arranged,and the CCSM cells hypertrophy,swell,distort,the nucleus also swollen round in the DEHP groups.By Masson staining,there are more CCSM cells in cavernous fiber around the cavernous sinus in the control group;And the CCSM cells are significantly reduced,but the collagen fibers increases obviously in the DEHP groups.By ICH staining,ER-beta,eNOS,caveolin-1 are mainly distributed on the inner wall of the cavernous sinus.Compared with controls,the expressions of ER-beta and the eNOS proteins reduce in the middle and high dose groups(P<0.05);But the expression of caveolin-1 increase in high dose group(P<0.05).By RT-PCR,the expression of eNOS mRNA in high dose group is reduced compared with control group(P<0.05),but caveolin-1 mRNA’s expression increase(P<0.05).In the high dose group,corpora cavernosa apoptotic cells increase compared with in the control(P<0.05).By Western blotting test,compared with the control group,the Bcl-2 protein’s expression reduce(P<0.05),but the bax protein’s expression increase in the high dose(P<0.05).Conclusion DEHP could induce ED of adolescent rats,possible mechanisms are:1.DEHP reduces the rat serum T and E2 levels;2.DEHP inhibits the expression of ER beta,eNOS,while enhances the expression of caveolin-1 in rat penis sponge cells,which also inhibits the eNOS’ activity;3.The DEHP do some effects on rats’ penile tissue structure,promotes the corpus cavernosum tissue fibrosis and cell apoptosis.The third part Explore the mechanism of tonifying kidney and excreting polluted on DEHP induced ED based on the theory of turbidity toxinObjective Observe tonifying kidney and excreting polluted on DEHP induced ED to discuss the therapeutic rules on turbidity toxin.Methods 30 rats are randomly divided into 5 groups,each group has 6,which respectively named,the control group,Clomiphene group,QiGui low,middle and high groups.All of them are filled with DEHP 100 mg/kg/d in the morning everyday,for 14 consecutive days.From the 8th day,each group is respectively filled with normal saline 2 ml/d,Clomiphene 2.25 mg/kg/d,QiGuiDiHuang prescriptions 12 g/kg/d,36 g/kg/d,108 g/kg/d in the afternoon to intervene,for 14 consecutive days.24 hours after the last treatment,indicators are tested(the same as part 2).Results In the Clomiphene group,the rats loss their gloss,which are unresponsive,occasionally fidgety,likely lying down,which has lost its weight compared with other groups at the end(P<0.05).Compared with control group,the erectile incubation time increases,the E2 levels increases,the T levels decreases,T/E ratio decreases too,the sponge cell apoptosis increases in the Clomiphene group.P of above<0.05.Meantime,the CCSM decreases,cell arrangement is irregular,the hypertrophy and swelling deformation CCSM cells increases,swelling round nuclei increases,and the collagen fiber increases.Compared with control group,the incubation time reduces,erection number increases,the level of T increases,T/E ratio increases too,the expression of ER-beta and eNOS protein increases,the expression of caveolin-1 reduces,the expression of eNOS mRNA increases,the expression of caveolin-1 mRNA reduces,the sponge cells apoptosis decreases,the expression of Bcl-2 protein increases,and expression of bax protein decreases in the middle dose group.P of above<0.05.Meantime,CCSM cells increases,cell arrangement is more regular;Hypertrophy and swelling deformation CCSM cells reduces,swelling round nucleus reduces,collagen fibers reduces too.Compared with Clomiphene group,erectile incubation time reduces,E2 levels are lower,level of T,T/E ratio are increased the group,and cell apoptosis are reduced,the expression of eNOS protein increases in the low,middle and high dose group,P above<0.05;But only in middle dose group,erectile times increases,the expression of ER-beta protein increases,the expression of caveolin-1 protein decreases,the expression of eNOS mRNA are increased,the expression of caveolin-1 mRNA decreases;The expression of Bcl 2 protein increases and the expression of bax protein decreases,P above<0.05.Conclusion Tonifying kidney and excreting polluted(QiGuiDiHuang prescription)could alleviate the ED induced by DEHP,possible mechanisms:1.QiGuiDiHuang prescription raises the level of serum T and T/E ratio;2.QiGuiDiHuang prescription increases the expression of ER beta and eNOS on endothelial cells,at the sametime,reduces the expression of caveolin-1,thus protects the vascular endothelial function;3.QiGuiDiHuang prescription protects rats penile tissue structure,inhibit the corpus cavernosum tissue from fibrosis and cell apoptosis.4.DEHP’s influence on hormones,vascular endothelial and the structure of CCSM cells conformed to the pathogenesis of Turbidity toxin,which injures Yin and Yang of the body,is weightier and viscous,causes qi and blood stasis,so that the blood is not enough in veins.In the end the penis loss glory and is out of use.5.Tonifying kidney and excreting polluted could invigorate the essence of kidney,balance the Yin and Yang of the body,nourish the Qi and blood to fill the vein,and nourish the penis,...