Mathison J C, Schreiber R D, La Forest A C, Ulevitch R J
J Immunol. 1983 Jun;130(6):2757-62.
The results from a number of clinical and experimental studies have suggested that during endotoxemia, suppression of adrenocortical steroidogenesis may occur. We have examined the possibility that macrophages are the source of a factor that suppresses adrenocortical steroidogenesis. Resident and peptone-elicited peritoneal exudate macrophages (PEM) from C3HeB/FeJ mice were incubated for 4 hr at 37 degrees C in the presence or absence of T cell hybridoma-derived lymphokine (LK) that contained high concentrations of MAF activity (assessed by induction of nonspecific tumoricidal activity in PEM). The LK was removed by rinsing, and fresh medium was added, followed by Salmonella minnesota R595 LPS (final concentration 10 micrograms/ml). After 18 hr at 37 degrees C the PEM supernatants and control medium from flasks without cells were harvested and stored at -20 degrees C. Explanted rabbit adrenocortical cells in 96-well plates were exposed to 30 microliters of PEM supernatant or control medium and ACTH (10 or 100 mU/ml) in a final volume of 120 microliters for 3 consecutive days. The adrenocortical cell supernatants were harvested each day, followed by replenishment of medium, PEM supernatant, and ACTH. Fluorogenic steroid production in wells that received control medium or supernatants from PEM not treated with LPS was normal (0.22 microgram +/- 0.010 (SD) per 5 X 10(4) cells). However, as much as 75 to 95% suppression of steroidogenesis was observed in wells that received supernatants from PEM treated with LK and LPS, compared to 40% suppression in wells that received supernatant from PEM treated with LPS alone. Continued exposure (over 3 days) of adrenocortical cells to supernatants from LPS-treated PEM resulted in progressively decreasing response to ACTH. Comparable suppressive activity was observed in supernatants from LPS-treated bone marrow-derived macrophages. In further experiments, suppression was observed in wells that were pretreated (22 hr) with the appropriate PEM supernatant, and evidence was obtained that the suppressive activity was not due to carry-over LPS. Finally, results from control experiments demonstrated that suppressive PEM supernatants neither inactivate ACTH nor interfere with the assay of fluorogenic steroids. Thus, these results suggest that during endotoxemia, products from LPS-stimulated macrophages may suppress adrenocortical function.
多项临床和实验研究结果表明，在内毒素血症期间，肾上腺皮质类固醇生成可能受到抑制。我们研究了巨噬细胞是否是抑制肾上腺皮质类固醇生成因子的来源。将来自C3HeB/FeJ小鼠的驻留型和蛋白胨诱导的腹腔渗出巨噬细胞（PEM）在存在或不存在含有高浓度巨噬细胞激活因子（MAF）活性（通过诱导PEM中的非特异性杀肿瘤活性评估）的T细胞杂交瘤衍生淋巴因子（LK）的情况下，于37℃孵育4小时。通过冲洗去除LK，加入新鲜培养基，然后加入明尼苏达沙门氏菌R595脂多糖（LPS，终浓度10微克/毫升）。在37℃孵育18小时后，收集来自无细胞培养瓶的PEM上清液和对照培养基，并储存在-20℃。将接种在96孔板中的原代兔肾上腺皮质细胞暴露于30微升PEM上清液或对照培养基以及促肾上腺皮质激素（ACTH，10或100毫单位/毫升）中，终体积为120微升，连续3天。每天收集肾上腺皮质细胞上清液，然后补充培养基、PEM上清液和ACTH。接受对照培养基或未用LPS处理的PEM上清液的孔中荧光类固醇生成正常（每5×10⁴个细胞0.22微克±0.010（标准差））。然而，与仅接受用LPS处理的PEM上清液的孔中40%的抑制率相比，接受用LK和LPS处理的PEM上清液的孔中类固醇生成的抑制率高达75%至95%。肾上腺皮质细胞持续暴露（超过3天）于LPS处理的PEM上清液导致对ACTH的反应逐渐降低。在LPS处理的骨髓来源巨噬细胞的上清液中观察到类似的抑制活性。在进一步的实验中，在用适当的PEM上清液预处理（22小时）的孔中观察到抑制作用，并且获得证据表明抑制活性不是由于残留的LPS。最后，对照实验结果表明，具有抑制作用的PEM上清液既不会使ACTH失活，也不会干扰荧光类固醇的测定。因此，这些结果表明，在内毒素血症期间，LPS刺激的巨噬细胞产生的产物可能抑制肾上腺皮质功能。
