| Objective: To investigate the effect of 900 MHz mobile phone frequency electromagnetic radiation on testicular oxidative damage and the expression of Prdx2 and Prdx4 protein in rats and and explore the mechanism of action of the Guilingji.On the basis of previous studies,the expression of testicular ultrastructure,antioxidant enzymes,peroxiredoxin 2(Prdx2)and peroxiredoxin 4(Prdx4)were the main research indicators.The Chinese medicine Guilingji was selected as the intervention factor to observe the effect of 900 MHz mobile phone frequency electromagnetic radiation on the testicular function of rats,and the possible mechanism of action of the Guilingji.Methods: Fifty healthy SD male rats were randomly divided into five groups,10 in each group: false radiation group,4h radiation group,8h radiation group,4h Chinese medicine group,and 8h Chinese medicine group.False radiation group radiations 0 hours,and the other four groups were exposed to 900 MHz electromagnetic radiation(370 ?W/cm2)for 4h and 8h,continuous radiation for 15 d,and the 4h Chinese medicine group and the 8h Chinese medicine group were irradiated with the Guilingji suspension.The other three groups were given distilled water.At the end of the experiment,the rats was sacrificed and take out the organizations.The pathological morphology of rat testis and the ultrastructural changes of testis were observed by HE staining.The changes of oxidative and antioxidant indexes in rat testis and rat serum were detected by TBA method.The expression of prdx2 and prdx4 protein was detected by immunohistochemistry and Western blot..Result:1.Comparison of general conditions of rats in each groupDuring the experiment,no significant change in response sensitivity was observed in each group.Daily intake of water is normal,The rats in the Guiling group had more stools.The skins of the rats in each group were intact and white and shiny.The body weight of the rats in each group increased,the difference was not statistically significant(P>0.05).2.Comparison of testicular coefficients of rats in each groupThere was no significant difference in testicular weight and testicular coefficient between the groups(P>0.05).3.Comparison the testicular histomorphology of rats in each groupIn the false radiation group,the seminiferous tubules of the testes of the rats were tightly connected,and the inner layers of the seminiferous tubules were structurally intact;It can be seen that spermatogonia,primary spermatocytes,secondary spermatocytes(occasionally),sperm cells are arranged neatly,and the lumen is filled with mature sperm;Compared with the false radiation group,the histological changes of the testis in the 4-hour radiation group were not significant.In the 8-hour radiation group,the testicular seminiferous tubules have a clear structure,and the spermatogenic cells at all levels are clearly distributed,but the microscopy is loose,and the number of mature spermatozoa in the small lumen is reduced.In the 4-hour Chinese medicine group,the layers of the testicular seminiferous tubules were clear and intact,and the seminiferous tubules were filled with mature spermatozoa;Compared with the 8-hour radiation group,the testicular tissue morphology of the rats in the 8-hour Chinese medicine group was basically normal.4.Comparison the testicular ultrastructure in rats of each groupCompared with the false radiation group,the spermatic membrane of the4 h radiation group was invaginated,the supporting cells were slightly separated from the basement membrane of the seminiferous tubule,the intercellular space was widened,some mitochondria were swollen and contained vacuoles,and the ridge of mitochondria are disappeared;Compared with the false radiation group,the thickening of the basement membrane of theseminiferous tubules in the 8h radiation group was thickened and the gap between the supporting cells was widened.The vacuoles between the cells and the cells were obvious.The spermatogonial cells showed irregular membranes,obvious depression,and mitochondrial dumbbell-shaped malformation.The mitochondrial part of the ridge disappears and contains vacuoles;Compared with the 4h radiation group,the basement membrane of the testicular seminiferous tubules in the 4h Chinese medicine group tends to be normal,the support cells are restored to tight junctions,the intercellular gap is reduced,and the mitochondrial structure is basically normal;Compared with the 8h radiation group,the testicular cells and intercellular space were reduced in the8 h Chinese medicine group,the basement membrane of the seminiferous tubules and the supporting cells became normal,and occasional vacuoles were observed in the cells,occasionally,mitochondria are swollen and vacuolated.5.Comparison of MDA,GSH content,SOD and POD activities in serum of rats in each groupCompared with the false radiation group,the serum GSH content and SOD activity of the rats in the 4h radiation group and the 8h radiation group were decreased significantly,and the MDA content was increased significantly(P<0.05).The serum POD activity of the 8h radiation group was decreased significantly(P<0.05).Compared with the 4h radiation group,serum MDA content increased significantly and SOD activity decreased in the 8h radiation group(P<0.05).Compared with the 4h radiation group,the MDA content in the serum of the 4h Chinese medicine group was significantly lower(P<0.05).Compared with the 8h radiation group,the serum MDA content in the8 h Chinese medicine group was decreased significantly,and the SOD activity was increased significantly(P<0.05).There was no significant difference in the change between other indicators.6.Comparison of MDA,GSH content and SOD and POD activities in testicular tissue of test in each groupCompared with the false radiation group,the SOD activity of the testis tissue was decreased significantly in the 4h radiation group,and the MDAcontent in the testis tissue of the 8h radiation group was increased significantly(P<0.05),and the GSH content,SOD and POD activities were significantly decreased(P<0.05);Compared with the 4h radiation group,the GSH content decreased in the 8h radiation group.Compared with the 8h radiation group,the MDA content in the testis tissue of the 8h Chinese medicine group was significantly decreased(P<0.05),and the GSH content and SOD activity were significantly increased(P<0.05).There was no significant difference between the other indexes(P>0.05).7.Comparison of Prdx2 and Prdx4 protein expression in testis of each group(immunohistochemistry)The false radiation group of Prdx2 and Prdx4 are expressed in spermatogonia,primary spermatocytes,secondary spermatocytes and sperm cells.The expression is gradually stronger from spermatogonial cells to sperm cells.Compared with the false radiation group,the expression of Prdx2 and Prdx4 in the radiation group significantly reduced,the expression of spermatogonia and primary spermatocytes,and the expression of secondary spermatocytes,sperm cells and sperm decreased.The average photometric values of Prdx2 and Prdx4 protein expression in the testis of the drug-administered group were significantly increased,especially in the Chinese medicine group.Compared with the false radiation group,the average luminosity values of Prdx2 and Prdx4 protein in the testis of the 4h radiation group and the 8h radiation group were significantly lower(P<0.05),and the expression of Prdx2 and Prdx4 protein in the testicular tissue of the 8h radiation group was lower than that in the 4h radiation group.The expression of Prdx2 and Prdx4 protein in testis tissue of 4h Chinese medicine group was significantly increased(P<0.05).Compared with the 8h radiation group,the expression of Prdx2 and Prdx4 protein was significantly increased in the 8h Chinese medicine group(P<0.05).8.Comparison of Prdx2 and Prdx4 protein expression in testis of each group(Western blot)To compared with the false radiation group,the average gray scale ratio of Prdx2 and Prdx4 protein expression in the testis of each group was significantly decreased(P<0.05).Compared with the 4h radiation group,the expression of protein in the 4h Chinese medicine group was increased significantly(P<0.05).Compared with the 8h radiation group,the expression of protein in the 8h Chinese medicine group was significantly increased(P<0.05).In addition,compared with the 4h radiation group,the expression of Prdx2 and Prdx4 protein in the testis tissue of the 8h radiation group was significantly decreased(P<0.05).9.Comparison of apoptosis in testis tissue of rats in each group.The spermatogenic cells were observed in each group,The nucleus is brownish yellow for positive expression.The false radiation group was mainly found in spermatogonia;the 4h and 8h radiation groups were found in spermatogonial cells,spermatocytes and sperm cells.In the 8h radiation group,some mature spermatozoa showed yellow-brown expression;The 4h Chinese medicine group was found in spermatogonia,occasionally in primary spermatocytes;the 8h Chinese medicine group was found in spermatogonia,primary spermatocytes,and occasionally in sperm cells.Conclusion:1.Although there is no significant abnormality in the morphology of rat testis induced by electromagnetic radiation at 900 MHz mobile phone frequency,the ultrastructural changes of testicular tissue in rats are obvious,and the changes of cell membrane and mitochondria is obvious,which can aggravate the oxidative damage of testicular tissue in rats,decreased the oxidase activity,and weakened the expression of Prdx2 and Prdx4 protein,and apoptosis in each group,which has a time-effect relationship.Rats and humans do not belong to the same genus,and oxidative damage may be related to the radiation dose,however,the results of this experiment have certain guiding significance for the clinic.humans can be made aware of the effects of radiation and prevent it.2.Guilingji can improve the damage of ultrastructure of rat testiculartissue induced by radiation,increase the expression of Prdx2 and Prdx4 in testis tissue,reduce oxidative damage.However,the in-depth mechanism of electromagnetic radiation damage to human tissues and organs has not been effectively confirmed,and further research is needed. |
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