摘要
目的探讨大肠埃希菌感染对雄性小鼠生殖细胞凋亡和P53蛋白表达的影响,从分子水平阐明清利生精丸治疗大肠埃希菌感染所致男性不育的机制。方法 50只雄性小鼠,经膀胱注射大肠埃希菌,建立动物感染模型,观察15d后随机分为5组:感染组(不治疗)、清利生精丸大、中、小等3种浓度治疗组(药物浓度分别为22.5、13.5和4.50g/ml)、呋喃坦啶治疗组,分别编码为MN、MTa、MTb、MTc、MTd组,另取10只小鼠膀胱注射生理盐水作为正常对照组(编码为CT)。各治疗组连续灌胃药液10d,应用流式细胞术检测小鼠睾丸生殖细胞凋亡率,免疫组化技术测定生殖细胞P53蛋白表达情况。结果各组小鼠均有一定的睾丸生殖细胞凋亡率,其中MN组生殖细胞凋亡率57.44%,CT组28.54%,MTa组30.11%,MTb组28.59%,差异有统计学意义(P<0.01);MN组生殖组细胞凋亡率与MTc(46.54%)和MTd组(43.41%)比较差异无统计学意义(P>0.05)。MN、CT、清利生精丸各治疗组和MTd组表达P53蛋白睾丸生殖细胞百分比差异有统计学意义(P<0.01)。结论大肠埃希菌感染小鼠后,可导致小鼠睾丸生殖细胞P53蛋白的表达上升,生殖细胞凋亡增加。清利生精丸通过下调P53蛋白,降低生殖细胞的凋亡率,提高小鼠生殖能力。因此,清利生精丸是治疗大肠埃希菌感染性不育的有效的药物之一。
Objective To study the effects Escherichia coli infection on the apoptosis of mice germ cells and expression of P53 protein in male mice and further to clarify the molecular mechanism of the Qing Li Sheng Jing Pill used to treat male sterility due to E.coli.Methods An animal infection model was established and fifty male mice were infected by injecting E.coli via the bladder.Mice were randomly divided into five groups（groups MN,MTa,MTb,MTc,and MTd,n=10） 15 days post-infection（dpi）：an untreated group,a treatment group given a high-dose Qing Li Sheng Jing Pill（22.5 g/ml）,a treatment group given a medium-dose Qing Li Sheng Jing Pill（13.5 g/ml）,a treatment group given a low-dose Qing Li Sheng Jing Pill（4.50 g/ml）,and a treatment group given Western medicine.Ten more mice were injected with saline via the bladder to serve as a normal control group（CT）.Infected mice were fed medicine 10 days continuously.The rate of germ cell apoptosis was detected by flow cytometry and the expression of P53 was immunohistochemically detected in testicular tissue;histopathological changes were also observed.Results The apoptosis of germ cells was detected in all infected groups and there was a significant difference between MN（57.44%） and CT（28.54%）,MTb（28.59%）,and MTa（30.11%）（P0.01）.The level of apoptosis decreased in MTc（46.54%） and MTd（43.41%） after treatment,but the difference was not statistically significant（P0.05）.The level of expression of P53 protein was significantly higher in MN than in CT,MTa,MTb,MTc,and MTd（P0.01）.Conclusion Results demonstrated that E.coli infection increased the rate of germ cell apoptosis and the level of P53 expression.Qing Li Sheng Jing Pills enhanced reproductive capacity by means of decreasing the level of P53 protein expression and the rate of germ cell apoptosis.Therefore,Qing Li Sheng Jing Pills may be an effective drug to treat infertility caused by E.coli.
出处
《中国病原生物学杂志》
CSCD
2011年第9期657-660,共4页
Journal of Pathogen Biology
基金
山东省计生委资助项目(No.2005-13)
关键词
清利生精丸
大肠埃希菌
动物模型
感染
生殖细胞凋亡
不育
雄性小鼠
Qing Li Sheng Jing pill
Escherichia coli
animal model
infection
germ cell apoptosis
infertility
male mice
