Antagonistic Effect Of Astragalus Injection On Reproductive Toxicity In Diesel Exhaust Particles Exposed Mice

Objective:With the expansion of urbanization and industrialization,China has become one of most seriously polluted countries in the world.Diesel exhaust particles(DEP) which contain polycyclic aromatic hydrocarbons,dioxins,nitrophenol etal,is the main component in air pollution.DEP owning chemicals activity of endocrine disrupting,can affect the expression of estrogen receptor and has activity of estrogen,anti-estrogen and anti-androgen. DEP was proved as bearing an obvious reproductive toxicity in animal experiments. Astragalus injection is extract of astragalus,its main compositions are amino,flavonoids, polysaccharidel and trace elements etal.Astragalus injection can antagonize the toxicity on motility of mice sperm and effect of ATP enzyme of cauda epididymidis induced by cadmium chloride,reduce and protect the apoptosis number of bilateral testicular germ cells in testicular torsion reset mice.Through this animal model:after ether anesthesia,the mice respectively were inhaled(through posterior pharyngeal) 30ml DEP suspension(12μg/μl, 3μg/μl,0.8μg/μl) or 30ml PBS to mimic inhalation of different concentrations of diesel exhaust by organism.Study the impact of DEP on reproductive capacity in ICR mice;and the antagonistic effect of astragalus injection on reproductive toxicity of ICR mice exposed in diesel exhaust particles.We aimed to investigate the density of sperm,total sperm activity,in vitro fertilization(IVF) rate,changes of epididymis and testicular structure.Strengthen the weak links in the research about the relationship between the male infertility and air pollution, investigate the mechanism of Chinese medicine in the treatment of male infertility.For the treatment of clinical patients with unidentified infertility,the experiment provided an experimental evidence and theoretical basis.Methods:60 mature(SW) male ICR mice were randomly divided into phosphate buffer group(group PBS),DEP(diesel exhaust particles ) exposed group(0.8μg/μl,3μg/μl, 12μg/μl DEP),DEP exposed and 10mg/ml astragalus injection treatment group,and diesel exhaust particles exposed and 20mg/ml astragalus injection group,6 in each group.The exposed model was established according to De Hennezel.After ether anesthesia,the mice respectively were inhaled(through posterior pharyngeal) 30ml DEP suspension(12μg/μl, 3μg /μl,0.8μg /μl) or 30ml PBS.Meanwhile exposed groups were injected with 1 ml 0.9%NaCl,intervention groups respectively were injected with 10mg/ml or 20 mg/ml astragalus injection everyday until the experiment was over.The male ICR mice were exposed twice weekly for four weeks,executed one week later.ICR male mice were ex executed by cervical dislocation.Removing the testis and cauda epididymidis in aseptic conditions.Then weight right testis and cauda epididymidis,and preserved as pathologic specimens.Take sperms from left cauda epididymidis.Detect relative testis weights,relative epididymis weights,sperm density,total activity,and in vitro fertilization(IVF) rate.Results:1 Sperm density,total activity,IVF rate,and thickness of spermatogenous epithelium and epididymis gland in 12,3μg/μl/mouse DEP- exposed groups decreased significantly,the relative epididymis weights increased significantly when compared with PBS group and 0.8μg /μl/mouse DEP-exposed group(P≤0.05).There is no statistical difference among the groups with regard to the relative testis weights(P＞0.05).Astragalus injection(10mg/ml, 20mg/ml) can significantly improve the detection indicators(P≤0.05)caused by being exposed to 3μg/μl,12μg/μl DEP suspension except the relative testis weights.But there is no difference between two concentrations.There is a positive correlation between the total sperm activity and the IVF rate(P=0.000).2 Histological examination of the testis:the wall of spermatogenous epithelium was thick;the layer of cell was wide and there were many sperms in arterial lumen in PBS and 0.8μg /μl/mouse DEP-exposed groups.The wall of spermatogenous epithelium became thinner significantly,spermatogenous epithelium arranged loosely;a few of atrophy seminiferous tubules can been viewed around the normal seminiferous tubule occasionally in 3.0μg/μl/mouse DEP- exposed group.The situation was much worse in 12.0μg/μl/mouse DEP-exposed group.The situation was improved in astragalus injection groups(10mg/ml,20 mg/ml)especially to 12,3μg/μl/mouse DEP-exposed groups.3 Histological examination of the epididymis:The epithelium cells of epididymis gland arranged closely,cilia of high columnar epithelium cells were clear and their margo libers were tidy.High columnar epithelium cells secreted vigorously.There were many ripe sperms in epididymis gland lumen in PBS and 0.8μg/μl/mouse DEP- exposed groups.The thickness of epididymis gland became thinner significantly,cilia were fuzzily,margo libers were unintelligble.The number of ripe sperms became fewer,necrosis cells can be seen in epididymis gland lumen in 3.0,12.0μg/μl/mouse DEP-exposed groups.The phenomenon was improved in astragalus injection groups(10mg/ml,20 mg/ml)especially to 12,3μg/μl/mouse DEP-exposed groups.Conclusion:1 Sperm density,total activity,IVF rate and thickness of spermatogenous epithelium and epididymis gland decreased significantly;the relative epididymis weights increased significantly and had bad impaction on the histological examination of the testis and epididymis in 3.0μg/μl,12.0μg/μl DEP groups;0.8μg/μ1 DEP group hadn't.2 Treating with 10mg/ml,20mg/ml astragalus injection can increase sperm density,total activity,IVF rate and thickness of spermatogenous epithelium and epididymis gland;decrease the relative epididymis weights;improve the condition of the histological examination of the testis and epididymis.But there is no difference between two concentrations.