| Objectives Previous studies have demonstrated that there is obvious harm function of long term application of rhubarb extract on uterus and ovary of rats,for further screening of rhubarb extract reproductivetoxicity.This study compared the rhubarb rhubarb water extract,chloroform,ethyl acetate,n-butanol extract and water extract on the ovarian and uterine index in rats,female glycol?E2?and progesterone?P?,testosterone?T?,estrogen receptor?ER?,progesterone receptor?PR?,effect of uterine and ovarian ultrastructure,determine the main effect of the extract of rhubarb on uterine and ovarian function and structure,lay the foundation for the further discussion of parts and components of rhubarb toxicity on reproductive system.Basics.Methods Followed by chloroform,ethyl acetate,the water extract was extracted by n-butanol,the corresponding extract and water soluble.60 SD female rats were randomly divided into normal control group,rhubarb extract group?4g/kg?,chloroform extract group?4.44mg/kg?,ethyl acetate extract group?40.96mg/kg?,is the butanol extract group?79.43mg/kg?,water extract group?835.72mg/kg?,each dose equivalent to 4 g/kg crude rhubarb,continuous intragastric administration for 60 days.After the last administration of 24 h in serum,uterus,ovary Calculate,uterus,ovary index;ELISA assay in rat serum P,T,E2 levels;immunohistochemical determination of ER in the uterus,ovary,the expression level of PR protein;HE staining,observe the uterine gland under the light microscope,the structure change of follicles in the ovary;observe the uterus under transmission electron microscope,ultrastructure of ovarian changes in the structure.Results 1 The effect of uterine and ovarian index:Rhubarb extract group,ethyl acetate extracts of rat ovarian index compared with the normal control group were decreased by7.5%,6%,the uterus index compared with the normal control group were in creased by18.3%,6%.n-butanol extract group,water extract group of rat ovarian index compared with control group were reduced by 19.7%,23.4%,the uterus index compared with the normal control group were decreased by 0.02% and 4.4%,but the differences were not statistically significant?P > 0.05?;chloroform extracts of rat ovarian index compared with normal control group in creased 4.6%,but the difference was not statistically significant?P>0.05?,uterine index and the normal control group in creased by 44.5%,the difference was statistically significant?P < 0.05?.2 Effect on the uterus,ovarian tissue structure:Rhubarb extract,ethyl acetate extract,n-butanol extract group,water extract group in rat serum P levels compared with normal control group were decreased by 2.34%,9.54%,9.25%,6.08%,but the differences were not statistically significant?P> 0.05?;the level of T in serum of rats with normal control group comparisons were reduced by 4.91%,4.24%,6.74%,4.25%,but the differences were not statistically significant?P>0.05?;chloroform extracts of rat serum P levels compared with normal control group decreased 19.49%,the level of T decreased compared with normal control group 18.90%,the differences were statistically significant?P<0.05?;rhubarb extract,chloroform extract,ethyl acetate extract group,n-butanol extracts of rat serum E2 levels were compared with the normal control group were in creased by 53.58%,87.78%,60.23%,59.28%,the differences were statistically significant?P < 0.05 or P < 0.01?;water soluble E2 levels in serum were compared with normal control group in creased12.93%,but the difference was not statistically significant?P>0.05?.3 Effects of ER?PR?levels on the uterus?endometrium and glands?and ovary?granulosa cells?: The chloroform extract,ethyl acetate extract group,n-butanol extracts of rat uterus in the expression level of ER compared with the normal control group were in creased by323.01%,117.77%,82.79%,the differences were statistically significant?P<0.05 or P<0.01?;rhubarb extract group and water soluble compounds in rat uterus the expression level of ER compared with the normal control group were increased by 44.07% and27.58%,there were no significant differences?P>0.05?.Rhubarb extract group,water extract group of rat uterus in the expression level of PR compared with the normal control group were increased by 19.15% and 43.75%,but the differences were not statistically significant?P>0.05?;chloroform extract,ethyl acetate extract and n-butanol extracts of rat uterus in the expression level of PR and normal control groups were increased by 99.74%,88.21%,84.92%,the differences were statistically significant?P<0.05 or P < 0.01?.The chloroform extract,ethyl acetate extract group rat ovary ER expression level compared with the normal control group were decreased by 60.88%,43.08%,the differences were statistically significant?P<0.05 or P<0.01?;water soluble group rat ovary ER expression level compared with normal control group increased7.52%,but there was no statistically significant difference?P > 0.05?;rhubarb extract group,n-butanol extracts of rat ovary ER expression level compared with the normalcontrol group were decreased by 30.99% and 30.46%,there were no significant differences?P>0.05?.The chloroform extracts of rat ovarian PR levels compared with normal control group increased 140.21%,the difference was statistically significant?P<0.01?;n-butanol extract group,rhubarb extract group rat ovary PR expression level compared with the normal control group were increased by 53.56% and 19.16%,there were no significant differences?P > 0.05?;ethyl acetate extract group,water extract group rat ovary PR expression level compared with the normal control group were decreased by 44.52% and 7.67%,there were no significant differences?P>0.05?.4 On the structure of uterus and ovary:Each extract group compared with the normal control group showed different degrees of uterine gland dilated,epithelial cells were flat,partial vacuolization or high columnar hyperplasia,to increase the number of nuclear increased.The number of blood vessels increased with cystic dilatation.Interstitial edema.The most obvious osteoporosis,chloroform extract group,acetic acid ethyl acetate extract group.Compared with the normal control group,the number of follicles in each group was decreased,the number of corpus luteum,the number of locked follicles increased,the number of granulosa cells in the ovary of chloroform extract group was reduced,and the arrangement of cells in the follicle was disordered.5 On the ultrastructure of uterus and ovary:Normal control endometrial epithelial microvilli rats rich glandular cells arranged orderly,rich,nuclear center,nuclear membrane was clear and intact,there are many secretory cells.The cytoplasm and cell organelles,more mitochondria,Golgi apparatus,endoplasmic reticulum,mitochondria cristae.Rhubarb decoction group rats endometrial microvillus shedding,gland cell membrane integrity,cell rupture,margination,organelles decreased.Chloroform extracts of rat uterine epithelial cells were almost completely absent,the gland cell number was significantly reduced,arranged in disorder,secretory cells decreased,the nucleolus and the chromatin dispersion,edge There were vacuoles in the stroma,and the cells were significantly reduced,and there was a gap between the cells.The ethyl acetate extract group was similar to that of the chloroform extract group.The n-butanol extract group and the water soluble group were close to the normal control group.l group in rat ovarian granulosa cells,the cells are large,round shape,complete nuclear center,nuclear membrane was clear and intact,chromatin distribution,abundant cytoplasm and secretory cells.Cell cell organelles,mitochondria are mitochondrial cristae clear,there are many Golgi and mitochondria.The lipid dropletsscattered.Nuclear margination appeared rhubarb decoction group in rat ovarian granulosa cells,granulosa cells and interstitial vacuoles,organelles decreased.The number of chloroform extracts of rat ovarian granulosa cells decreased,disordered cell nuclear rupture,chromatin margination,mitochondria decreased,edema,vacuolar,mitochondria There was a gap between the cells,and the ethyl acetate extract group was close to that of the chloroform extract.The n-butanol extract and water soluble group were close to the normal control groupConclusions Rhubarb extract used in this experiment and various extracts of uterus,ovarian function and structure have a certain degree of damage,the damage degree is the most significant chloroform extract,extract the main hints of chloroform extract produced the reproductive toxicity of rhubarb.Description of rhubarb on uterine and ovarian toxicity of parts and components are the chloroform extract. |
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