Dhydroepiandrosterone (DHEA) is a steroid hormone produced by the adrenals that serves as a precursor for many steroid hormones.The aim of this work is to study the effects of DHEA on liver, ovary, breast and skin of adult and senile female albino rats and the role of vitamin E as a protective agent.The period of this experimental study extended for 6 months. Both adult and senile female albino rats were received the medications for 4 months to study the toxic effects of the drug then the rats stayed without any medication for another 2 months to study the degree of regression in these effects if present.Six hundred female albino rats were used in this study (300 adult and 300 senile). Each of them was equally divided into 5 groups, the first three groups were control groups receiving distilled water, corn oil and vitamin E respectively, the last two groups were received DHEA and combined DHEA and vitamin E treated groups.The studied dose of DHEA in both adult and senile female albino rats was 300mg/kg B.W/day in a maximum therapeutic dose, vitamin E was given in a dose of 100mg/kg B.W/day.Twenty rats from each group in both adult and senile female albino rats were sacrificed after 2& 4 months of daily drug administration and after 2 months follow up without any drugs (6 months). Specimens obtained from liver, ovary, breast and skin were subjected to histopathological examination using H&E stain and AgNORs hisochemical stain which confirm the proliferative changes seen by light microscopy. The mean values of the mitotic count indicated by the number of nuclear dots in a certain number of cells in AgNOR stain of each organ were estimated by Computerized Analysis System 200 (CAS 200).(A) Histopathological study:Throughout the whole period of the study, histopathological changes obtained from the three control groups (adult and senile) were within normal.Histopathological Effect on Liver:(I) DHEA treated group:After 2 months of daily DHEA administration, histopathological changes in the liver were in the form of congestion, inflammation and hepatocellular carcinoma. Congestion was present in 90% of rats in adult female albino rats, and in 45% of rats in the senile group. Inflammation occurred in all rats 100% of the adult group and in 25% of rats of the senile group. As regarding HCC, it was not present in adult rats, but was present only in 20% of rats of the senile group.After 4 months, of daily DHEA administration in adult group inflammation regressed and was evident only in 30%, congestion in 85 % of rats while HCC were detected in 40 %of rats. In the senile group, progression occurred in all histopathological changes evident by the occurrence of congestion in 60% of rats, inflammation in 55% of rats and HCC manifestation in 75% of rats.After 6 months, (when rats stayed for 2 months without any medications as a follow up period) liver congestion and inflammation showed regression in both adult and senile groups, but no regression in HCC changes. In adult group, congestion was present in 10% of rats and inflammation in 5% of rats, while manifestations of HCC were still detected in 40% of rats. In senile group, congestion and inflammation were detected in 15% of rats, while HCC changes were observed in 65% of rats. This means that DHEA cause hepatic toxicity in senile rats more than in adult.(II) Combined DHEA and vitamin E treated group:After 2 months of this combination therapy, congestion was evident in 90% of rats liver and inflammation in 70% of rats, while no manifestation of HCC could be detected in adult female rats.In senile female rats, this combination therapy regress both liver congestion and inflammation to 35% and 25% respectively, while HCC changes was present only in 10% of rats.After 4 months of combined DHEA and vitamin E treatment in adult group, there was a regression in both liver congestion and inflammation (45% & 10% respectively), while HCC changes appear in 35% of rats. In senile group, congestion decrease to 15% of rats and inflammation to 10% of rats, while HCC changes increased to 65% of rats.After 2 months follow up period with cessation of combined DHEA and vitamin E treatment, there was a regression in both liver, congestion and inflammation in adult group (10%) of rats, while HCC manifestations were obvious in 35% of rats. In senile rats, liver congestion was observed in 10% of rats and inflammation in 5% of rats, while HCC changes appear in 60% of rats. This means that vitamin E had a protective role in liver congestion and inflammation but no role in HCC of both adult and senile liver.Histopathological Effect On Ovary(I) DHEA treated group:After 2 months of daily DHEA administration, cystic changes were detected in 40% in ovary of adult group, while they were detected only in 10% of senile rat group.After 4 months of daily DHEA administration, ovarian cystic changes showed significant progression and were observed in all the adult rats 100% and 40% in senile rats group.After 6 months (2 months of DHEA cessation), these cystic changes did not show significant regression as it represents 85% in adult rats group and 30% in senile rats group indicating that there was no improvement in these cystic changes after this follow up period.The detectable significant histopathological changes were cystic changes in the form of polycystic ovary in both adult and senile rats all over the period of the study.(II) Combined DHEA and vitamin E group:After 2 months of daily administration of this combined therapy, cystic changes of the ovary were detected in 35% of adult rats group, while showed only in 5% of senile rats group.After 4 months of treatment with DHEA & Vitamin E, cystic changes progressed significantly into 90% in adult rats and in 35% in senile rats.After 6 months (2 months without this combined therapy), cystic changes did not regress as it is represented in 70% of adult rats and in 20% of senile rats.Vitamin E did not offer any protection to ovarian cystic changes in both adult and senile female rats throughout the whole period of the study.Histopathological Effect On BreastThe detectable significant histopathological changes were glandular hyperplasia in the mammary tissue.(I) DHEA treated group:After 2 months of daily DHEA administration, 35% of the adult female rats showed glandular hyperplasia of the breast tissues, while it is showed in 20% of the senile rats.After 4 months of daily DHEA administration, glandular hyperplasia was progressed to be detected in 95% of adult female rats and 75% of senile female rats.After 6 months, (when DHEA treatment stopped for 2 months) glandular hyperplasia was detected in 40% of adult rats group and 45% of senile rats group.(II) Combined DHEA and vitamin E treated group:Vitamin E offered a protective role against DHEA induced glandular hyperplasia of senile rats breast, while no protection could be detected in adult rats breast.After 2 months of daily administration of DHEA with vitamin E, only 15% of adult rats group and 20% of senile rats.After 4 months of this combined therapy, glandular mammary hyperplasia was detected in 75% of adult rats group, while was detected in 20% of senile rats group indicating the protective role of vitamin E in senile female rats against glandular hyperplasia.After 6 months (2 months follow up without this combined therapy) glandular hyperplasia was detected in 30% of adult rats group, while was detected only in 15% of senile rats indicating the protective role of vitamin E in senile female rats even after cessation of vitamin E with DHEA.Histopathological Effect On SkinIn DHEA treated group, there was an obvious improvement in the morphological appearance of the senile rats skin in the form of increased the skin thickening and the size and number of the sebaceous glands, but these changes were not detected in adult female rats. These beneficial effects of DHEA on senile rats skin were augmented through Co-administration of DHEA with vitamin E.After 2 months of daily DHEA therapy, skin thickening was detected in 40% of DHEA treated rats group while in combined DHEA and vitamin E treated group 60% of rats showed skin thickening. Increase in size and number of sebaceous glands occurred in 50% of DHEA treated senile rats and in 70% of combined DHEA with vitamin E treated senile rats.After 4 months of therapy, significant improvement in skin thickening was detected in both DHEA 65% and combined DHEA and vitamin E 90% treated senile rats. Also, hyperplasia of sebaceous glands was detected in 60% of DHEA treated group and 90% of combined DHEA and vitamin E treated group.After 6 months (2 months follow up), skin thickening remained in high incidence even after stoppage of DHEA treatment (55%) or combined DHEA and vitamin E (65%), but there was a regression in sebaceous glands hyperplasia after cessation of DHEA treatment (20%) which not regressed in the size and number in senile rats after cessation of combined DHEA and vitamin E treatment (55%).(B) Cell proliferation by AgNORs stain and its estimation by Compeuterized Analyzer System 200 (CAS 200):Proliferative changes detected in histopathological changes of each organ (liver, ovary and breast) was confirmed by AgNORs stain and the mean value of the mitotic count was accurately estimated by CAS 200 through counting the number of dots inside the nuclei in a certain number of cells.The mean values of the mitotic count of control groups were within normal throughout the period of the study in both adult and senile.Effect On Liver(I)DHEA treated group:After 2 months of DHEA treatment in adult female rats, the mean value of the mitotic count was (2.115 ± 0.374) that progressed after 4 months to (5.471 ± 3.431) and still high after 6 months when DHEA treatment stopped for 2 months (5.039 ± 0.39).In senile group, the mean values of the mitotic count inside liver cells were higher than those inside liver cells of adult group as the mean value after 2 months was (4.305 ± 2.338) that progressed after 4 months to (6.706 ± 4.285) and remained high even after stoppage of the drug for 2 months (5.908 ± 3.356).This means that there was no improvement in HCC manifestation after this follow up period after cessation of DHEA in liver of both adult and senile female albino rats.(II) Combined DHEA and vitamin E treated group:After 2 months of combined DHEA and vitamin E treatment in adult female rats, the mean value of the mitotic count inside liver cells was (1.980 ± 0.207) that progressed after 4 months to (4.956 ± 2.434) and still high after 6 months ( 4.995 ± 1.357) when DHEA treatment stopped for 2 months.In senile group, the mean values of the mitotic count inside liver cells were higher than those inside liver cells of adult group as the mean value after 2 months was (3.728 ± 1.716) that progressed after 4 months to (6.169 ± 4.140) and still high even after cessation of this combined drugs for 2 months (5.552 ±3.396).This means that there was no protective role of vitamin E in HCC after this follow up period in liver of both adult and senile female albino rats.The proliferative changes in the liver cells was manifested by hepatocellular carcinoma (HCC) which early detected by the results of CAS 200 (2 months) and progressed after 4 months to remain high even after cessation of the drug for 2 months (6 months). These values were higher in senile rats than those in adult rats in both DHEA and combined DHEA and vitamin E treated group. No role of vitamin E in the protection against HCC.Effect On Ovary(I) DHEA treated group:After 2 months of daily DHEA administration in adult female rats, the mean value of the mitotic count was (6.980 ± 3.38) which progressed after 4 months to (9.019 ± 4.837) and remained high after 6 months even with cessation of DHEA treatment for 2 months (8.635 ± 3.83).In senile group, the mean value of the mitotic count was (4.7 ± 1.52) which progressed to (6.896 ± 4.147) and remained high even after stoppage of the drug for 2 months (6.03 ± 2.7).(II)Combined DHEA and vitamin E treated group:After 2 months of this combined therapy in adult female rats group, the mean value of the mitotic count was (6.412 ± 2.73) which progressed after 4 months to (8.75 ± 3.33), then after cessation of the drugs for 2 months the value was (8.7 ± 2.5).In senile group, the mean value of the mitotic count after 2 months was (4.55 ± 1.0) which progressed after 4 months to (6.5 ± 1.1) and remained high after cessation of drugs (5.92 ± 0.3).The proliferative activity in the ovarian cells was in the form of polycystic ovary, this proliferation starts at the end of the first period of the study and progressed after 4 months. After cessation of the drugs either DHEA alone or combined DHEA and vitamin E, there were no regression in the ovarian cystic changes, thus the mitotic count remained high after 6 months. No role of vitamin E in protection against ovarian cystic changes.Effect On BreastThe proliferative activity in the breast tissues in the form of glandular hyperplasia was confirmed by AgNORs stain and the mean value of the mitotic count in the nuclei of the breast cells was estimated by CAS 200. The lesion was progressed after 4 months then regressed after cessation on the drug in both adult and senile female rats in both DHEA and combined DHEA and vitamin E treated group.There was a protective role of vitamin E against DHEA induced glandular hyperplasia in senile female albino rats after 4 and 2 months follow up period (6 months), while no protective role for vitamin E in adult female breast throughout the periods of the study.(I) DHEA treated group:After 2 months of daily DHEA administration in adult female rats, the mean value of the mitotic count was (3.250 ± 2.085) which progressed after 4 months to (4.640 ± 2.650) then regressed after 6 months (2 months follow up without treatment) to (3.591 ± 1.57).In senile group, with daily administration of DHEA, the mean value of the mitotic count in breast cells was (1.828 ± 0.599) which progressed after 4 months to (3.645 ± 1.476) then regressed after 6 months (2 months follow up without treatment) to (2.025 ± 0.542).(II) Combined DHEA and vitamin E treated group:After 2 months of daily DHEA with vitamin E administration, the mean value of the mitotic count was (2.925 ± 0.547) which progressed after 4 months (4.020 ± 2.986) then regressed to (3.039 ± 0.396) after the follow up period.In senile group taking this combined therapy, no significant difference in the mean value between the three periods of the study. The mean value of the mitotic count was (1.656 ± 0.590) after 2 months, (1.619 ± 0.707) after 4 months and (1.661 ± 0.372) after cessation of the drugs.Effect Of SkinThe mean values of the mitotic count were within normal values throughout the whole periods of the study in both adult and senile female albino rats either taking DHEA or combined DHEA and vitamin E.CONCLUSION(1) The effect of DHEA in liver of senile rats is more hazardous than in liver of adult rats.(2) Dehydroepiandrosterone (DHEA) as a precursor for sex hormone had toxic effects on organs under hormonal control like ovary and breast which were more obvious in adult than in senile female albino rats especially with prolonged administration of high doses.(3) When DHEA was stopped for 2 months as a follow up period, a regression was detected in liver congestion and inflammation of both adult and senile female rats, also there was a regression in glandular hyperplasia of breast tissue of both adult and senile female albino rats. But no regression was detected in the toxic effects of DHEA regarding HCC and polycystic ovaries in both adult and senile female albino rats.(4) Vitamin E protected the liver against the effect of DHEA induced toxic capillarites (congestion and inflammation). However, it did not protect liver against hepatocellular carcinoma in both adult and senile female albino rats.(5) Vitamin E had no protective role in DHEA induced polycystic ovaries (PCO) in both adult and senile female albino rats.(6) Vitamin E protected the breast tissue against DHEA induced glandular hyperplasia only in senile female albino rats.(7) DHEA had a real benefit effects on the skin of senile female rats through improving the thickness of the skin as well as the size and number of the sebaceous glands.(8) Concerning skin improvement by DHEA administration, there was a regression in the improved sebaceous glands when DHEA was stopped for 2 months as a follow up period.(9) Vitamin E augmented the effect of DHEA on the skin of senile female rats by increasing the skin thickening and the size and number of the sebaceous glands. Also, preserve the size and number of these glands even after cessation of combined DHEA and vitamin E.(10) AgNORs stained slides estimated by Computerized Analyzer System 200 (CAS 200) was an early and sensitive indicator for cell proliferation which observed in the histophatological picture.