The Study Of Compound Angelica Injection To Interfere Toxicity On Reproduction Cell Of Female Mice Exposed To Diesel Exhaust Particles

Objective:Through the observation of mice models which were polluted by diesel exhaust particles (DEP),the research intends to analyze DEP's influence on oocyte growth,clarify its mechanism,and investigate the relationship between DEP and female reproductive toxicity. After confirming that DEP induce female reproductive toxicity,the research continues to investigate the likely preventive effect of compound Angelica injection and its mechanism. This will provide a theoretical basis for clinical application of Traditional Chinese Medicine in protection of reproductive function,and consequently promote the generalization and application of Traditional Chinese Medicine.Method:The research object was 273 21-day-old healthy ICR mouse,which were divided into 13 groups.Beside normal control group and normal-treatment group,low dose DEP model, middle dose DEP model,high dose DEP model,DEP treatment-model group(DEP middle dose and high dose),middle dose model with low dose traditional Chinese medicine,middle dose model with middle dose traditional Chinese medicine,middle dose model with high dose traditional Chinese medicine,high dose model with low dose traditional Chinese medicine, high dose model with middle dose traditional Chinese medicine,high dose model with high dose traditional Chinese medicine.When they were 21-31 days old,the mouse in the former 9 groups would be injected with DEP liquids which are provided by Public Health College of The Ohio State University at pharynx posterior wall by sample pipettor,once every three days for ten days.To Chinese medicine treatment groups,they would be injected with 5g pharmacognostic/kg·d,10g pharmacognostic/kg·d,or 20g pharmacognostic/kg·d of compound angelica into abdominal cavity after being intoxicated,and normal-treatment group,DEP treatment-model group(DEP middle dose and high dose) would be injected with physiological saline 0.3ml/10g·d.Then the mouse would be observed on following aspects:general situation,body weight,ovaries weigh and organ coefficient.We observed the effects of diesel exhaust particles on mice oocyte maturation in vivo and in vitro fertilization,observed oocyte's ultrastructural under electron microscope,measured the contents of SOD,MDA and GSH in ovaries tissue bomogenate and measured the contents of oocyte mitochondria DNA(mtDNA) by means of real time PCR technique.Results:Model Group1.In each group of mice,it was not significantly different in body weight change(P＞0.05).The ovarian wet weight and ovarian organ coefficient of DEP in the high-dose exposure group versus control group and DEP's low dose group were significantly lower (P＜0.05); 2.By electron microscopy's findings:the mitochondria of oocytes in DEP's middle and highdose exposure groups swelling and vacuolar degeneration,but the ultramicrostructure of cell in low dose group were normal;3.DEP the middle-and high-dose exposure in mice ovarian tissue SOD content,GSH content significantly decreased,MDA content increased significantly(P＜0.05);4.All experimental groups of oocytes did not affect germinal vesicle breakdown,DEP's middle-and high-dose exposure group can significantly reduce the mouse(0 h) oocytes first polar body release rate(P＜0.01 ),and increased oocytes degenerate rate(P＜0.01); The fertilization rate of oocytes at 24 and cleavage rate of embryos after 2-cell stage at 48h of in vitro fertilization,when the total rate with the control group,DEP's low-dose group decreased significantly(P＜0.01);5.Compared with the control group,DEP3.0ug/ul group and mice DEP12.0ug/ul oocyte mtDNA copy number(transformed into logarithms) and the control group,DEP0.8ug/ul there is statistical difference between groups study significance(P＜0.01). Compound angelica medicine treatment group:DEP mouse exposed to varying degrees the situation improved,as demonstrated by:1.In each group of mice,there was no significant difference in body weight change(P＞0.05).In compound angelica high-dose treatment group ovarian wet weight and ovarian organ coefficient with the corresponding dose of DEP's model group has significantly increased(P＞0.01);2.By electron microscopy's findings:compared with model group,the ultramicrostructure of cell on the ovaries appear improve in compound angelica treatment group;3.In compound angelica high-dose treatment group,mouse ovarian tissue SOD content and GSH content was significantly increased,MDA content was decreased significantly(P＜0.05);4.All experimental groups of oocytes did not affect germinal vesicle breakdown,compound angelica high-dose treatment group can be significantly increased in mouse(0 h) oocytes first polar body release rate(P＜0.01),and reduced oocytes degenerate rate(P＜0.01).The fertilization rate of oocytes at 24 and cleavage rate of embryos after 2-cell stage at 48h of in vitro fertilization,when the total dose rate and the corresponding DEP the model group increased significantly(P＜0.01);5.For mouse oocyte mtDNA copy number(after the transformation of logarithms),of compound angelica high-dose treatment group with the corresponding dose of DEP compared with the model group,the difference had statistical significance(P＜0.01).Conclusion:1.The acutely exposured to diesel exhaust particles of animal models,which have been successfully made in the research are propitious to the study of how DEP and compound Angelica affect female genital system.2.DEP is reproductively toxiferous,and it will do harm to the ovaries and oocyte separately. The probability of in vitro fertilization decreases with the increasing contents of DEP in mice.These prove that DEP is harmful to conception. 3.From the view point of Traditional Chinese Medicine,DEP which is one of the factors of environmental pollution,was treated as a kind of toxin.It could affect the embryo and pregnant mother,resulting in dysfunction of organ,imbalance of Yin-yan,and damage of reproductive system which could lead to abortion.4.Some Traditional Chinese medicine can counteract DEP's toxicity.The compound Angelica can relieve the damage of genitals and oocyte of mouse,and achieve the effect of "strengthening vital Qi and eliminating pathogens".So when properly used,the compound Angelica is effective to the prevention and cure of DEP exposure.