去氫表雄固醇 (DHEA)是腎上腺皮質所分泌的類固醇荷爾蒙，是製造性荷爾蒙的中間產物，在哺乳動物體內參與多種生理作用，具有調節免疫反應的功能。本實驗室先前曾以離體狀態，研究DHEA對老鼠免疫細胞製造腫瘤壞死因子 (TNF-alfa) 及轉型生長素 (TGF-beta) 的影響，發現DHEA能促進P388D1細胞及腹腔巨噬細胞製造TNF-alfa 及 TGF-beta。本研究則延續前人研究結果，欲知老鼠施打DHEA後，對體內免疫細胞製造TNF- alfa及TGF- beta的功能，是否造成影響。Balb/c老鼠皮下施打DHEA 72 小時之後，採取血清並取出腹腔巨噬細胞，腹腔巨噬細胞以酯多醣(LPS)刺激活化，培養特定時間後，收集培養基上清液並以酵素免疫分析法測定培養基上清液及血清中TNF- alfa 及TGF-beta 的濃度。結果顯示，細胞在離體培養下，TNF- alfa 的分泌量在6小時達到最高峰，而老鼠施打每克重50ug及100ug的DHEA，其腹腔巨噬細胞分泌TNF- alfa 的能力顯著增加，但高於100ug 之劑量後，TNF- alfa 的分泌量即回復至控制組的水準。本實驗也以RT-PCR確認DHEA對TNF- alfa 的影響，結果顯示DHEA可促進老鼠腹腔巨噬細胞TNF- alfa 基因的表現。採取施打DHEA老鼠之血清，化驗結果顯示，DHEA不顯著影響血清中TNF-alfa 的濃度。由施打DHEA的老鼠身上分離之腹腔巨噬細胞，進行離體培養24小時後，無論施打何種劑量，其分泌TGF-beta 的能力均與控制組無顯著差異。採取施打DHEA老鼠的血清進行化驗，發現老鼠施打每克重100ug的DHEA時，血液中TGF-beta的濃度顯著低於控制組。dexamethasone是一種很強的免疫抑制劑，由施打dexamethasone的老鼠身上分離之腹腔巨噬細胞，進行離體培養6小時，其分泌TNF-alfa的能力顯著降低，不過如果同時施打dexamethasone及DHEA，則DHEA會對dexamethasone產生拮抗作用，即減少dexamethasone對巨噬細胞功能的抑制能力。分析血清中TNF-alfa 的含量，雖然dexamethasone的抑制效果不明顯，但是同時施打DHEA時，仍對dexamethasone的作用有影響。反之，dexamethasone顯著促進血清中TGF-beta 的濃度，然而DHEA無法拮抗dexamethasone對血清中TGF-beta濃度的促進作用。由本研究結果推知，施打DHEA對腹腔巨噬細胞分泌TNF-alfa的能力有顯著的促進作用，對血清中TGF- 的濃度有抑制作用。但在活體內，DHEA對分泌此兩種細胞激素的影響可能受到其他因素的干擾以致TNF-alfa的血清濃度 及腹腔巨噬細胞分泌TGF- beta的能力未因為施打DHEA而改變 ; 此外，dexamethasone及DHEA雖然同為固醇類荷爾蒙，但是DHEA可拮抗dexamethasone對TNF-alfa 的抑制效應。

Dehydroepiandrosterone (DHEA) is a predominant androgen secreted by adrenal cortex. DHEA has been proposed to play an important role in regulating physiological and immunological system in mammals. Our previous report suggested that DHEA caused an enhancement in TNF-alfa and TGF-beta production by both P388D1 cells and peritoneal macrophage under in vitro condition. The specific aim of present study is to investigate whether DHEA have ability to regulate TNF-alfa and TGF-beta production under in vivo condition. Balb/c mice administrated DHEA subcutaneously were sacrificed at 72 hrs after the treatment, then the serum and peritoneal macrophages were collected. The peritoneal macrophages were cultured with LPS for a period of time and the supernatant was then collected. Both TNF-alfa and TGF-b concentration were quantified using ELISA kits. Results suggested that the TNF-alfa secretion by peritoneal macrophages from the mice injected with 50 ug per g body weight and 100 ug per g body weight of DHEA were significantly increased and reached peak 6 hr after incubation. However, the amount of TNF-alfa secretion by macrophages returned to control level when the dose higher than 100 ug per g body weight was given. Result from RT-PCR analysis confirmed the finding that expression of TNF-alfa mRNA was enhanced by in vivo administration of DHEA. By contrast, the serum level of TNF-alfa was not significantly affected by the drug treatment. The TGF-beta secretion by peritoneal macrophages under in vitro culture did not show a significant difference from mice receiving carrier (100 % ethanol). However, the serum level of TGF-beta of mice injected with 100 ug per g body weight of DHEA was significantly lower than the mice received carrier. Dexamethasone is a potent immunosuppressor. Mice injected 1ug per g body weight of dexamethasone showed a significant reduction in TNF-alfa secretion by peritoneal macrophages. However, injection of DHEA together with dexamethasone could overcome the immunosupressive effect of dexamethasone on TNF-alfa secretion. The serum level of TNF-alfa did not show a significant change in dexamethasone treated mice but it showed a significantly increase in the mice received both DHEA and dexamethasone. Mice received dexamethasone showed a significant increase in the serum level of TGF-beta. However, DHEA could not antagonize the augmented effect of dexamethasone on the serum level of TGF-beta. In conclusion, mice injected DHEA potentiated the TNF-alfa production by peritoneal macrophages but decreased the serum level of TGF-beta. Since the effect of DHEA on both TNF-alfa and TGF-beta production might be interfered by other factors under in vivo condition, the effect of DHEA on the serum level of TNF-alfa and the TGF-beta was not significant. In addition, DHEA might be an effective antagonist to overcome the suppressive effect of dexamethasone on TNF-alfa secretion in vivo.

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