| ObjectiveToday,male infertility affects more than 20 million Patients in the world and has become one of the main Problems affecting male reproductive health.Teratozoospermia is one of the common manifestations of male infertility.At Present,the Pathogenesis is not clear,the factors involved are extensive and complex,and the effect of drug treatment is average.Some Patients need to implement assisted reproductive technology to achieve the purpose of fertility.About 8%-12% of couples in the world suffer from infertility,and among these couples,40%-50% of them are caused by "male factors".And semen abnormalities are the most common,accounting for at least about 1/3 of abnormal sperm.Whether the sperm morphology is normal is the key to distinguish between sperm and egg.Whether sperm can undergo normal fertilization in the body depends entirely on whether the sperm has a normal shape.The Pathogenesis of sperm abnormalities is very complicated,and there are many influencing factors,including the following: chromosomal abnormalities,gene mutations and deletions,hormone disorders,immune abnormalities,infections,deterioration of the surrounding environment,toxic radiation,varicocele,Reactive oxygen species(ROS)appear excessive,oxidative stress and sperm DNA damage are mainly related to sperm motility.Relevant studies believe that many harmful factors have caused the activation of the signal Pathway of the sperm surface Protein receptor.Based on the signal transduction method,the normal structure of the sperm can be changed,and eventually the sperm movement may appear abnormal morphology.Among them,most of the current research hot topics focus on ROS excess,oxidative stress and sperm DNA damage.However,there is no unified understanding of the signal Path for Process information transmission.As a classic nuclear transcription signal transduction Pathway,JAK/STAT signal transduction Pathway can regulate all life activities of cells such as normal growth,development,differentiation and apoptosis.Recent studies in recent years have found that the JAK2/STAT3 signaling Pathway can be based on mitochondrial-mediated oxidative damage activities,thereby realizing the life activity of regulating cell apoptosis;at the same time,by activating and controlling the JAK2/STAT3 signaling Pathway,the damage response can be adjusted as well as Apoptosis of cells.In addition,if a STAT3 inhibitor is used in vitro,it will cause a decline in sperm motility,and finally lead to a decline in fertilization ability.However,whether the JAK2/STAT3 signaling Pathway is related to the morphology and function of sperm,and whether changes in the JAK2/STAT3 signaling Pathway will cause changes in the morphology and function of sperm is still unknown.Therefore,the in-depth study of the relationship between the JAK2/STAT3 signaling Pathway and sperm morphology and function will greatly help to understand the mechanism of sperm morphology and function abnormalities,and achieve the clinical purpose of guiding male infertility and sperm abnormalities.Traditional Chinese medicine has a long history of treating infertility in China for more than 2,000 years.Male infertility and sperm abnormalities belonged to the concept of "childless" in ancient times.The mechanism of this condition is quite complicated,and kidney deficiency and blood stasis are one of the key Pathological factors of this disease,which can be divided into kidney deficiency syndrome and blood stasis syndrome.There are various factors that can lead to insufficient kidney yang,low energization,deficiency of cold,lack of warmth,sperm injury,or kidney yin deficiency,yin deficiency,hyperactivity,semen suffering,sperm burns or insufficient qi,but it cannot Promote blood circulation,yang Deficiency,warm blood vessels,slow blood flow,blood stasis,blood flow obstruction,it is easy to accumulate Pathological Products,destroy the living environment of sperm survival,and ultimately increase the Probability of abnormalities,affect the normal function of sperm,and cause infertility.For clinical treatment,needle-Pick therapy can improve the microenvironment for sperm survival,increase blood flow,resist ischemia,hypoxia,and oxidative stress,thereby Protecting sperm from damage and treating abnormal sperm infertility.This article uses acupuncture therapy to treat male infertility of kidney deficiency and blood stasis,using computer-assisted semen analysis(CASA),sperm morphology staining,and flow cytometry(FCM)to detect sperm's conventional Parameters,morphology and Whether the sperm DNA is missing,the Pregnancy rate of the Patient's spouse should also be tracked.The purpose is to exlore the analysis of acupuncture treatment on semen Parameters,sperm morphology,sperm DFI and Pregnancy effects of infertility Patients.In addition,based on Western blot technology(WB),the relative expression levels of JAK2 and STAT3 Proteins in normal and abnormal spermatozoa can be compared and monitored,and finally the relationship between sperm morphology and JAK2/STAT3 Protein expression levels can be deeply grasped.In order to understand the effects of acupuncture treatment on semen Parameters,sperm morphology and JAK2/STAT3 Protein expression levels,CASA,sperm morphology staining and WB were selected to detect changes in semen Parameters,sperm morphology and JAK2 and STAT3 Protein expression levels before and after treatment.Finally,different concentrations of JAK2/STAT3inhibitor(AG490)were used to treat normal sperm.At the same time,based on the effects of sperm morphology staining,flow cytometry to detect sperm morphology and sperm DFI Performance,grasp the effect of JAK2/STAT3 signaling Pathway on sperm morphology and sperm DFI,for in-depth research and analysis of male infertility and sperm The Pathogenesis of deformity and clinical treatment Provide reliable evidence.MethodsThe research in this article will be carried out from two aspects: clinical research and experimental research:1.Clinical researchA total of 80 Patients were collected from the First Affiliated Hospital of Jinan University(Guangzhou Overseas Chinese Hospital)from March 30,2017 to September 30,2018.(Guangzhou overseas Chinese hospital).Due to the Poor efficacy of traditional western medicine treatment such as vitamin E and anti-oxidation,a total of 80 cases of Patients visited the hospital.At the same time,it is necessary to calculate the Percentage of normal sperm,the Percentage of abnormal sperm,the abnormal sperm index(TZI),the sperm deformity index(SDI),the head defect rate,the defect rate of the neck and middle,and the defect rate of the main segment of the tail.And the excess rate of residual cytoplasm;the method of density gradient centrifugation is used to extract sperm,and the WB method is used to detect the Protein expression level of JAK2 and STAT3.Method: Forty Patients with abnormal sperm were randomly divided into group A and group B.Group A(40 cases): Tonic of Needle-Pricking was given after dialectical addition and subtraction for a Period of 12 weeks(use the national Patent for needle Picking invented by Professor Chen Dong from the new needle Picking department of the first affiliated hospital of Jinan university);Group B(40 cases in total): Insist on giving the Patient 100 mg of natural vitamin E orally every day,twice a day,the whole course of treatment is 12 weeks.The semen Parameters of the two groups were analyzed before treatment and 12 weeks after treatment,and many characteristics such as total sperm count,sperm concentration,total motility and sperm motility Parameters were obtained.After 12 weeks of follow-up,the Pregnancy outcome of the spouses of the two groups of Patients was observed in time,and the clinical Pregnancy rate,miscarriage rate and live birth rate were also observed.2.Experimental research(1)The random sampling method was used to collect Patients who were in the First Affiliated Hospital of Jinan University(Guangzhou Overseas Chinese Hospital)from March 30,2017 to September 30,2018,40 people.And the Percentage of normal spermatozoa,Percentage of abnormal spermatozoa,abnormal sperm index(TZI),sperm deformity index(SDI),Percentage of head defects,neck and middle segments were calculated.Due to the Poor efficacy of traditional western medicine treatment such as vitamin E and anti-oxidation,a total of 80 cases of Patients visited the hospital.Symptoms: Patients with syndromes of kidney deficiency and blood stasis were included in the study as subjects(abnormal spermatozoa group);Men with the same age at the same age as male specialists in the Department of Reproductive Medicine were selected.(Normal spouses have a history of Pregnancy within the Past 3months.),normal semen detection in 20 cases(normal sperm group).The two groups of subjects used CASA to analyze semen routine Parameters of semen and obtain total sperm count,sperm concentration,total sperm motility,and sperm motility Parameters.Diff-Quik method was used to detect sperm morphology,and the Percentage of normal morphological spermatozoa and Percentage of abnormal spermatozoa were calculated.Sperm abnormality index(TZI),Sperm deformity index(SDI),Percentage of head defects,Percentage of neck and mid-segment defects,Percentage of defect at the main Part of the tail;Sperm density gradient centrifugation was used to extract sperms and extract them.Total sperm Protein was used to detect the expression of JAK2 Protein and STAT3 Protein by WB method.(2)Randomize the above-mentioned abnormal spermatozoa group of 40 Patients;into the Needle-Pricking group(A group)and no treatment group(B group);A group(40 cases): To Needle-Pricking treatment after dialectical addition and subtraction oral,the course of treatment is 12 weeks(use the national Patent for needle Picking invented by Professor Chen Dong from the new needle Picking department of the first affiliated hospital of Jinan university);group B(40 cases): blank group,no drug treatment;And the Percentage of normal spermatozoa,Percentage of abnormal spermatozoa,abnormal sperm index(TZI),sperm deformity index(SDI),Percentage of head defects,neck and middle segments were calculated.subjects of both groups were subjected to CAS analysis of semen routine Parameters before and after 12 weeks(treatment)to obtain total sperm count,sperm concentration,spermatozoa Total vigor and sperm movement Parameters;Diff-Quik method was used to detect sperm morphology,and the Percentage of normal spermatozoa,Percentage of abnormal spermatozoa,abnormal sperm index(TZI),sperm deformity index(SDI),Percentage of head defects,neck and middle segments were calculated.Percentage of defects,Percentage of main tail defects,and Percentage of residual residual cytoplasm.Sperm density gradient centrifugation was used to extract spermatozoa and extract total sperm Proteins.WB method was used to detect the expression of JAK2 Protein and STAT3 Protein,respectively.(3)Randomly collected 8 males(with normal Pregnancy history in the Past 3months)from the March 30,2017 to September 30,2018 in the male reproductive health department of the first affiliated hospital of Jinan university(Guangzhou overseas Chinese hospital).From 23 to 45 years old,normal spermatozoa were included in the study;routine collection of sperm,after treatment by density gradient centrifugation,different concentrations(0,25,50,100 ?mol/ L)JAK2/STAT3 inhibitor AG490 and sperm in vitro After incubation for 3 hours,sperm morphology was measured by Diff-Quik method,and the Percentage of normal spermatozoa was calculated.The sperm DFI was detected by FCM.Results1.Clinical research:Percentage of abnormal sperm morphology,and sperm forward movement(PR)after treatment in the Needle-Pricking treatment group were higher than before treatment.The sperm DFI,Percentage of head defects,Percentage of tail defects,and excess cytoplasmic residual rate were higher.Before treatment,the difference was statistically significant(P<0.05).After treatment of Needle-Pricking treatment group,the TZI,SDI,and neck defect rate were lower than before treatment.And the Percentage of normal spermatozoa,Percentage of abnormal spermatozoa,abnormal sperm index(TZI),sperm deformity index(SDI),Percentage of head defects,neck and middle segments were calculated.Semen volume and sperm The concentration and sperm count were higher than those before treatment,but the difference was not statistically significant(P>0.05).The normal sperm morphology after treatment in the vitamin E group was higher than before treatment,and the tail defect rate was lower than before treatment.The differences were statistically significant.Significance(P<0.05).Patients treated with TZI,SDI,sperm DFI,semen volume,total sperm count,head defect rate,neck defect rate,and excess cytoplasmic residual rate after treatment Before treatment,sperm concentration,total sperm motility,and sperm forward movement(PR)were higher than before treatment,with no statistically significant difference(P>0.05);Patients without treatment did not receive treatment.After 12 weeks,the excess cytoplasmic residual rate of the tail was lower than that of the 12 th week.The difference was statistically significant(P<0.05).The normal sperm morphology,semen volume,total sperm motility,head defect rate,and neck defect rate Increased,sperm abnormality index(TZI)and abnormal sperm index(SDI)did not change,sperm DFI,sperm concentration,total sperm count and sperm forward movement(PR)decreased compared with before treatment,the difference was not statistically significant(P> 0.05).2.Experimental results:(1)JAK2 and STAT3 were expressed in normal spermatozoa and teratozoospermia.JAK2 and STAT3 were highly expressed in healthy sperm in the spermatozoa group;JAK2 Protein and STAT3 Protein were expressed in teratozoospermia.The teratozoospermia group was significantly weakened;the difference in Protein expression between the two groups was statistically significant(P<0.05).The Percentage of normal sperm morphology was Positively correlated,abnormal sperm index and sperm abnormality index were negatively It is closely related to the expression of JAK2 and Stat3,the difference was statistically significant(P<0.05);Not related and the difference was not statistically significant(P>0.05).(2)JAK2 Protein and STAT3 Protein were expressed in the Needle-Pricking group and the untreated group;And the Percentage of normal spermatozoa,Percentage of abnormal spermatozoa,abnormal sperm index(TZI),sperm deformity index(SDI),Percentage of head defects,neck and middle segments were calculated.the expression of JAK2 Protein and STAT3 Protein in the Needle-Pricking treatment group was higher than that in the untreated group,and the expression level of the Protein between the two groups was statistically significant(P<0.05);Two groups of Patients in the Needle-Pricking treatment group and the non-treatment group were compared by using self-contrast method.The Percentage of normal sperm morphology,sperm motility,PR,JAK2 Protein and STAT3 Protein expression after treatment in the Needle-Pricking group were higher than those before treatment.Statistically significant(P<0.05).Semen volume and sperm concentration were higher than before treatment,TZI,SDI were lower than before treatment,and there was no significant difference(P>0.05);The Percentage of normal sperm morphology,semen volume,sperm concentration,sperm motility,PR,TZI in the untreated group.The SDI and STAT3 Protein expression was lower than before treatment,with statistically significant difference.(P<0.05).(3)The Percentage of normal sperm morphology and sperm DFI after incubating with AG490 and using AG490 for 3 hours showed that the normal morphology Percentage of sperm decreased with the increase of AG490 concentration,and the DFI of sperm increased with the increase of AG490,Both were statistically significant(P<0.05);however,there was no correlation between normal sperm morphology and sperm DFI by mean correlation analysis(r=0.24,P=0.14).Conclusions1.Needle-Pricking treatment can improve the normal sperm morphology of sperm in Patients with male infertility deformity sperm,reduce sperm DFI,reduce the Percentage of defects in the sperm head and the Percentage of tail defects,and increase the spouse's Pregnancy rate.2.For the normal sperm group and the abnormal sperm group,the expression of JAK2 and STAT3 Protein in the normal sperm group was higher than that in the abnormal sperm group.At the same time,the expression levels of JAK2 and STAT3 Protein in the abnormal sperm group were significantly decreased,and the abnormal sperm group JAK2 The expression level of STAT3 Protein was significantly lower than that of the normal sperm group.Needle Picking can increase the rate of normal sperm morphology,which may be related to the increase of JAK2 and STAT3 Protein expression and the activation of JAK2/STAT3 signaling Pathway.3.AG490 is a JAK2/STAT3 inhibitor,which can reduce the Percentage of normal sperm morphology and increase sperm DFI.The decrease in the Percentage of normal sperm and the increase in sperm DFI may be related to the inhibition of JAK2/STAT3 signal transduction Pathway. |
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