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Objective@#To investigate the effect of Ethephon on the testis pathological structure and apoptosis of spermatogenic cell in offspring male rats.@*Methods@#Twenty-four healthy female SD rats of 45 days old were randomly divided into control group, low-dose Ethephon group, medium-dose Ethephon group and high-dose Ethephon group according to body weight.The male rats of the same age were selected to mate with female rats.The rats were fed with Ethephon solution of different concentrations or 9 g/L saline every day, and they were continued to be fed with Ethephon during pregnancy and lactation.At the age of 7 days and 14 days, 10 offspring male rats were randomly selec-ted from each group and were put to death.The testicular tissue was stained with HE, and the morphological changes in the testis were observed with light microscope; the apoptotic cells were labeled with terminal deoxynucleotidyl transfe-rase dUTP nick-end labeling (TUNEL method) and the apoptosis index(AI) of testis spermatogenic cells was detected with fluorescence microscope.@*Results@#At the age of 7 days, the testis internal structure of the control group developed well, and the spermatic tubules were neatly and compactly arranged.In the low-dose Ethephon group, the seminiferous tubules of the testis were slightly smaller and the spermatogenic cells were loosely arranged compared with the control group.In the medium-dose Ethephon group, the testis seminiferous tubules were slightly disordered and the cell gap increased.In the high-dose Ethephon group, the testis development was poor, the diameter of seminiferous tubules decreased significantly, and the spermatogenic cells arrangement was in disorder.There was no statistically significant difference in spermatogenic cell AI between the low-dose group [(0.54±0.10)%] and the control group[(0.53±0.09)%] (P>0.05), while the spermatogenic cell AI in the medium-dose Ethephon group [(0.63±0.11)%] and the high-dose Ethephon group [(0.81±0.06)%] were higher than that in the control group, thus there exists a statistically significant difference (all P<0.01). The spermatogenic cell AI in the low-dose Ethephon group [(0.54±0.10)%] was lower than that in the medium-dose Ethephon group [(0.63±0.11)%], and the difference was statistically significant (P<0.05). The spermatogenic cell AI in the medium-dose Ethephon group was higher than that in the high-dose Ethephon group, and the difference was statistically significant (P<0.01). At the age of 14 days, the spermatogenic cells AI in control group, low-dose Ethephon group, medium-dose Ethephon group and high-dose Ethephon group were (0.54±0.08)%, (0.65±0.11)%, (0.77±0.11)%, and (0.88±0.10)% respectively, and the spermatogenic cells AI in all groups increased gradually, in which the differences were statistically significant (all P<0.01).@*Conclusions@#Excessive dose of Ethephon can induce pathological changes in testicular tissue and increase the apoptosis of spermatogenic cells, resulting in low fertility of offspring rats.
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Objective To investigate the effect of Ethephon on the testis pathological structure and apoptosis of spermatogenic cell in offspring male rats.Methods Twenty-four healthy female SD rats of 45 days old were randomly divided into control group,low-dose Ethephon group,medium-dose Ethephon group and high-dose Ethephon group according to body weight.The male rats of the same age were selected to mate with female rats.The rats were fed with Ethephon solution of different concentrations or 9 g/L saline every day,and they were continued to be fed with Ethephon during pregnancy and lactation.At the age of 7 days and 14 days,10 offspring male rats were randomly selected from each group and were put to death.The testicular tissue was stained with HE,and the morphological changes in the testis were observed with light microscope;the apoptotic cells were labeled with terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL method) and the apoptosis index(AI) of testis spermatogenic cells was detected with fluorescence microscope.Results At the age of 7 days,the testis internal structure of the control group developed well,and the spermatic tubules were neatly and compactly arranged.In the low-dose Ethephon group,the seminiferous tubules of the testis were slightly smaller and the spermatogenic cells were loosely arranged compared with the control group.In the medium-dose Ethephon group,the testis seminiferous tubules were slightly disordered and the cell gap increased.In the high-dose Ethephon group,the testis development was poor,the diameter of seminiferous tubules decreased significantly,and the spermatogenic cells arrangement was in disorder.There was no statistically significant difference in spermatogenic cell AI between the low-dose group [(0.54 ± 0.10)％] and the control group[(0.53 ±0.09) ％] (P ＞ 0.05),while the spermatogenic cell AI in the medium-dose Ethephon group [(0.63 ± 0.11) ％]and the high-dose Ethephon group [(0.81 ± 0.06) ％] were higher than that in the control group,thus there exists a statistically significant difference (all P ＜0.01).The spermatogenic cell AI in the low-dose Ethephon group [(0.54 ±0.10) ％] was lower than that in the medium-dose Ethephon group [(0.63 ± 0.11)％],and the difference was statistically significant (P ＜ 0.05).The spermatogenic cell AI in the medium-dose Ethephon group was higher than that in the high-dose Ethephon group,and the difference was statistically significant (P ＜0.01).At the age of 14 days,the spermatogenic cells AI in control group,low-dose Ethephon group,medium-dose Ethephon group and high-dose Ethephon group were (0.54 ± 0.08) ％,(0.65 ± 0.11) ％,(0.77 ± 0.11) ％,and (0.88 ± 0.10) ％respectively,and the spermatogenic cells AI in all groups increased gradually,in which the differences were statistically significant (all P ＜ 0.01).Conclusions Excessive dose of Ethephon can induce pathological changes in testicular tissue and increase the apoptosis of spermatogenic cells,resulting in low fertility of offspring rats.
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Objective To investigate the effects of Ethephon on spermatogenic cells and sex hormones levels of adolescent male rats.Methods The male SD rats of 25-day old were randomly (from the random number table) divided into high dose group (2 000 mg/kg),middle dose group (1 000 mg/kg),low dose group (500 mg/kg) and control group (the same amount of physiological saline).They were given Ethephon through stomach for 14 d.The pathological changes in testis tissues were observed by HE staining.The apoptosis of germ cells were detected by terminal transferase labeling (TUNEL).The automatic chemical luminescence immunoassay analyzer was used to test the serum sex hormone levels.Results After ethephon and saline lavage for 14 days in the dose of 2 000 mg/kg,1 000mg/kg,500 mg/kg concentrations respectively,body weight growth was significantly decreased (F =3.58,P =0.03).Testis mass growth [(0.91 ± 0.17) g,(1.13 ± 0.15) g,(1.21 ± 0.11) g,(1.29 ± 0.28) g] was significantly decreased (F =4.31,P =0.02).Experimental group spermatogenic cell apoptosis,the apoptosis index increased (F =156.00,P =0.00),high dose group[(2.40 ±0.18)％] and middle dose group[(1.72 ±0.14)％] was significandy increased (P ＜ 0.01).The levels of testosterone and estrogen in serum showed a decreasing trend along with the increase of the doses,and there was a statistical significance (F =11.85,38.93,all P =0.00).Compared with the control group [(0.86 ± 0.10) μg/L],the testosterone levels in high dose group [(0.31 ± 0.08) μg/L],middle dose group [(0.36± 0.05) μg/L] decreased significantly (P ＜ 0.01).Compared with the control group,low dose group,middle dose group [(36.43 ± 3.57) ng/L,(38.62 ± 2.24) ng/L,(31.87 ± 5.78) ng/L],the estrogen level in high dose group[(27.39 ± 2.11) ng/L] was significantly reduced (P ＜ 0.01).Conclusion Ethephon has reproductive toxicity,and can cause the serum level of testosterone and estradiol decreased,resulting in spermatogenic cell apoptosis index increased and the spermatogenic capability decreased.
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Objective To investigate the effect of arsenic(As2O3) on spermatogenic cell apoptosis and the expression of telomerase activity in adult rats.Methods 40 healthy male Sprague-Dawle rats were randomly divided into four groups and they were treated with As2O3 at doses of 0(control group),0.375,0.75 and 1.5 mg/kg body weight respectively through gavage for 16 consecutive weeks.The numbers of testicular sperm head were counted and the coefficient of testicular viscera,daily sperm production(DSP) were calculated in every group.The apoptosis of germ cell was assessed by in situ terminal deoxynucleotityl transferase mediated dTUP nick end labeling(TUNEL) technique.The activity of telomerase in the spermatogenic cells was determined by immunohistochemistry.Results In the testes of As2O3-treated rats,the coefficient of testicular viscera,the number of testicular sperm head and DSP in moderate and high dose groups decreased significantly than that of control(P
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In the occupational production and the daily life,the pollutant of pesticides may enter our bodies through the skincontact,the breath and the gastrointestinal absorption and so on. It may cause the reproductive organ and the spermatogenic cell damaged,abnormal endocrine function and the reproduction related genes expressed abnormally,resulting to abnormal spermatogenesis,which leads to oligospermia,asthenospermia,teratospermia and azoospermia. In order to look for a normal pattern and investigate the intrinsic mechanism of the cytotoxicity to the spermatogenic cell correlated with the pesticide pollution,it is necessary to review the relationship between the toxicity of pesticide pollutant and the abnormity of male spermatogenic cell,which will provide some basic information for preventing and controlling the reproductive toxicity of pesticides and ensuring the reproduction healthy of people.
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Objective To study the effects of manganese on the expression of caspase-3 and cytochrome-c in the spermatogenic cells of rats. Methods Forty-eight male SD rats were randomly divided into the high dose,low dose exposure groups and the control groups,16 in each,were given manganese(MnCl2?4H2O) at 15,30 mg/kg and normal saline,I.P. once a day, 5 times a week,for 6 consecutive weeks. In the 4th and 6th weekend,the testis were collected,the expression of caspase-3 and cytochrome-c in the testis was determined by immunohistochemical methods (SABC). Results The caspase-3-positive-cell rate in the spermatogenic cells of rats in the high dose and low dose exposure groups increased significantly (P
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Fluorescent staining method is a sensitive cytochemical staining method, it can reflect minimal changes of biochemical components of cells. The 7690-Xu fluorescent staining solution prepared with thioflavine can show the heterogeneo- us fluorescence of cells in different stages of differentiation. The younger cells with low degree of differentiation show blue fluorescence, the lower the degree the deeper the color. Along with the differentiation of cells, the blue color of fluorescence deflects to the red color of spectrum gradually, the blue color of fluorescence of cytoplasm changes gradually to bluish, bluish-green, and finally yellow color. The blue color of fluorescence of nucleus changes through greyish- blue to orangeyellow or orange-red.The cytoplasm of cells in various organs with high degree of differentiation appears yellow, and the nucleus orange-yellow or orange-red in color.
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Objective To investigate the development of neonatal mouse testis in castrated immunodeficient mice by monitoring the graft survival and weight and observing the structure of seminiferous epithelium and the composition of spermatogenic cells in grafts. Methods Neonatal Kun-ming mouse testis were grafted under the skin of castrated nude mice(7-12week-old).Grafts were then taken out at different time intervals(namely 16 time points: 3 days,1-11 weeks respectively and 3-6 months respectively).The survival rate of grafts was calculated and the wet weight was measured.Histological analysis was performed for the observation of the structure of seminiferous epithelium and the composition of spermatogenic cells in grafts. Results Four hundrcd and five grafts recovered out of 450 testis grafted, resulting in a recovery rate of 90.0%.The graft weight increased more than 40 times.The developmental pattern of seminiferous tubules and the appearance time of various spermatogenic cells in grafts were similar as seen in intact mice.Eight weeks after the grafrting,an increasing degradation of seminiferous epithelium was observed.Conclusion When neonatal mouse testis were grafted into nude mice,the developmental course was similar as that in normal donors.The optimal retrieval time of round spermatids and sperms was around the end of the first spermatogenesis wave,about 5-7weeks after the grafting procedure.