Clinical And Experimental Research On The Treatment Of Diabetic Erectile Dysfunction By Huoxue Tongluo Qiwei Decoction And "Leech-Centipede"

BackgroundDiabetes mellitus(DM)is one of the three major chronic diseases,which recognized by the World Health Organization.Diabetes-induced erectile dysfunction(DIED)is one of the common complications of diabetes,its incidence is high and it is closely related to cardiovascular diseases.As early as 1992,the National Institute of Health expert meeting determined that DM is one essential factor of sexual dysfunction in men and women.The incidence of ED in patients who suffer from DM is 1.9-5 times than those who do not suffer from DM,and the incidence of DIED reported worldwide is 20%-75%.As the country with the highest prevalence of diabetes worldwide,China is expected to reach 40 million patients who suffer from DIED by 2025.DIED belongs to the range of vascular ED,and its pathological basis is closely related to the damage of penile vascular endothelial function,which means that,DM mainly causes ED by affecting endothelial function.Thus,an in-depth study of vascular endothelial function and its possible mechanisms in a diabetic internal environment will help to further elucidate the pathogenesis of diabetic angiopathy,which could contribute to discover new targets in the treatment of DIED,and to provide reliable evidence about the safty and efficacy of the therapeutic drugs.PurposeThis study could be divided into clinical part and experimental part,which aims to further clarify the clinical efficacy of "Huoxue Tongluo Qiwei Decoction" combined with low-dose tadalafil in the treatment of DIED patients,besides,to explore the possible mechanism of"leech-centipede",which represent the method of "Huoxue Tongluo"in treating the model rats of DIED,in order to provide a theoretical basis for subsequent clinical treatment.MethodClinical studyA randomized controlled group was used.Patients who match the inclusion criteria of type 2diabetes erectile dysfunction would be recorded with general information,symptoms,signs,treatment history,and combined disease,which relate to the relevant TCM syndrome and these syndrome scores would be recorded.All the 66 patients were treated with Huoxue Tongluo Qiwei Decoction Formula Granules orally.The course of treatment was 28 days.After 28 days of treatment,the effective rate of IIEF-5 scores and TCM syndrome scores should be recorded before and after treatment.Besides,all kinds of adverse reactions during medication should also be recorded in detail.All we talked above aim to observe the clinical effect of Huoxue Tongluo Qiwei Decoction on patients who suffer from diabetes erectile dysfunction,as well as the change of IIEF-5 score and TCM symptom score.Experimental study50 male Sprague-Dawley rats were randomly and equally divided into blank control group,model control group,positive control group,Chinese medicine high-dose group,and Chinese medicine low-dose group.The blank control group was treated with deionized water for 8 weeks,the model control group was treated with 55mg/kg of STZ once,and 10 weeks after STZ injection,the viable diabetic rats would be screened out.The blank control group and the STZ model group were sequentially treated with deionized water.The positive control group was treated with 0.5 mg/kg/d of tadalafil(equivalent to 6 times of human dose).Chinese medicine high-dose group was treated with 0.6mg/kg/d "leech-centipede" which was made into granular suspension(equivalent to 12times of human dose),Chinese medicine low-dose group was treated with 0.15mg/kg/d "leech-centipede" which was made into granular suspension(equivalent to 3 times of human dose),all groups would accept intervention for 8 weeks.What we need to observe from the rat model of DIED,treated by "leech-centipede",contain the general condition,blood glucose level,pathomorphology of cavernous endothelial cells,ultrastructure of penile tissue,and cavernous vascular endothelial cell injury markers(Lectin like oxLDL Receptor-1,LOX-1),endothelial cell-associated adhesion molecules(sE-selectin,sICAM-1)and the effects of the expression of nNOS,eNOS,NOS,PDE5 protein and mRNA in penile tissue.ResultClinical studyThe improvement of IIEF-5score in the treatment group was better than that in the control group.The difference between the two groups was statistically significant,P<0.05.The improvement of TCM syndrome total score in the treatment group was better than that in the control group after treatment,which also exist statistical significance,P<0.05;The total effective rate was 81.8%in the treatment group,by contrast,51.5%in the control group,the difference about the effective rate between the two groups was statistically significant,P<0.05;There were 4 cases of adverse reactions,including 2 cases of headache in the treatment group,by contrast,there was 1 case of facial flushing,and 1 case of mild low back pain in the control group.The above symptoms were mild and could be spontaneously relieved without any special treatmentExperimental study1.After drug intervention,the general state and body weight of the DIED model rats among these groups did not show any significant difference,P>0.05;As for the blood glucose level,it seems that the blood glucose level of Chinese medicine group was better than that of the model group and the tadalafil group,however no significant difference was expressed in blood glucose levels among these groups,P>0.05;As for the penile erection frequency,it was lower in the high-dose group,the low-dose group and the tadalafil group than that in the blank control group,but it was higher than the model group,and the difference was statistically significant,P<0.05;2.In the serum test,the expression of LOX-1,sE-selectin and sICAM-1 in the serum of the model group was higher than that in the blank group,the difference was statistically significant,P<0.05;Comparing the high dose-group,the low-dose group and the tadalafil group with the model group,the expression of LOX-1,sE-selectin and sICAM-1 in these groups were lower,the difference was statistically significant(P<0.05),besides,this kind of inhibitory effect was more remarkable in the high-dose group,and the difference was statistically significant,P<0.05.3.In the observation of the ultrastructure of penile tissue,the distribution of sinusoids in the group of rats treated with the drug pair and the group of rats treated with tadalafil was relatively regular when comparing with the model group,besides,the density of endothelial cells was increasing,and erythrocytes were observed in some blood sinus spaces.In addition,through the observation of transmission electron microscopy,the plasma membrane of the endothelial cells in the model group was damaged and swollen,and the organelles were significantly reduced or even no organelle,and the nuclear chromatin concentration and edge accumulation was seen.After drug intervention,a certain degree of endothelial proliferation was observed in the drug-pair group and the tadalafil group.In addition,the structure of nucleus in these groups were intact,and the organelles such as mitochondria,Golgi apparatus and endoplasmic reticulum increased significantly,or were basically normal.4.We detected the expression of the protein through western blot way,no difference was detected in the expression of eNOS protein in each group.As for the protein expression of nNOS,the model group was lower that in the blank control group,besides,the drug-pair group and the tadalafil group were higher than the model group,and the differences above were statistically significant,P<0.05,and the high-dose group was superior to the low-dose group and the tadalafil group;The expression of NOS protein in the model group was lower than that in the blank control group,this expression in the drug-pair group and the tadalafil group were higher than that in the model group,the difference was also statistically significant,P<0.05,and the high-dose group was superior to the low-dose group.As for the expression of PDE5protein in the penile tissue,it was lower in the model group than that in the blank group,and the difference was statistically significant,P<0.05.There was no signifi cant difference between the low-dose drug-pair group and the model group,P>0.05,but the expression level of the high-dose drug-pair group was significantly higher than that in the tadalafil group and the model group,P<0.05;5.We detected relative mRNA through RT-PCR,and it showed that there was no significant difference in the expression of eNOS-mRNA in the DIED model rats.As for the expression of nNOS-mRNA,,the model group was lower than the blank control group,the drug-pair group and the tadalafil group were higher than the model group,the differences above were statistically significant,P<0.05,and the drug-pair high-dose group was better than the low-dose group and the tadalafil group.As for the expression of NOS mRNA,it was lower in the model group than that in the blank control group,and this kind of expression in the drug-pair group and the tadalafil group were higher than that in the model group,the differences were statistically significant,P<0.05,and the expression in the high-dose group was better than that in the low-dose group.As for the expression of PDE5 mRNA in penile tissue,the expression level in the model group was lower than that in the blank group,and the difference was statistically significant,P<0.05.There was no significant difference in the expression level between the low-dose group and the model group,P>0.05,but there was a significant increase in the expression level between the high-dose group,the tadalafil group and the model group,P<0.05.There was coherence between related mRNA and protein in the expression of them.Conclusion1.Huoxue Tongluo Method,which is represented by Huoxue Tongluo Qiwei Decoction,can safely and effectively improve the erectile function and systemic symptoms of patients who suffer from diabetes-induced erectile dysfunction;2."Leech-centipede" medicine-pair can increase the frequency of erection in rats with diabetes-induced erectile dysfunction and improve the erectile state,besides,improve the expression of serum LOX-1,sE-selectin,and sICAM-1 levels,among which the effect of the high-dose group is more obvious;3.When the"leech-centipede"drug-pair is in a high-dose state.It can promote the expression of nNOS or NOS proteins,besides it could lead to the promption of the expression of relative mRNA.The specific mechanism may be to inhibit the activity of PED5,which make it possible to achieve a PDE5 inhibitor-like effect.