Olubadewo Joseph O, Spitzer Judy A
College of Pharmacy, Xavier University of Louisiana, 1 Drexel Drive, New Orleans, LA 70125, USA.
Alcohol. 2003 Nov;31(3):137-47. doi: 10.1016/j.alcohol.2003.09.004.
We examined how acute diabetes mellitus and acute ethanol intoxication modulate factors that mediate immune responses as a basis for explaining the increased susceptibility to infection in these two conditions. Our working hypothesis is that ethanol intoxication in diabetes compromises host defense mechanisms to a greater extent than observed in each condition alone. Male and female rats were made diabetic with streptozotocin (65 mg/kg, i.p.). Forty-eight hours after administration of streptozotocin, rats either received no treatment (control group) or were treated with (1) ethanol (bolus injection of 1.75 g/kg, followed by a 3-h infusion at the rate of 300 mg/kg/h), (2) lipopolysaccharide [(LPS); 0.9 mg/kg], or (3) a combination of LPS+ethanol. At the end of 3 h, rats were killed, and the livers were digested by perfusion with collagenase-containing Hanks' balanced salt solution to isolate hepatocytes and Kupffer cells. To measure chemokine generation, hepatocytes (2.5x10(5) cells per well) and Kupffer cells (1x10(6) cells per well) were cultured for 20 h, and the supernatant was used to measure cytokine-induced neutrophil chemoattractant (CINC) and regulated on activation, normal T-cell expressed and secreted (RANTES) chemokines. Phagocytosis by Kupffer cells was measured by flow cytometry and expressed as mean channel fluorescence intensity (MCF). Induction of diabetes as well as treatment of nondiabetic rats with LPS, ethanol, or LPS+ethanol caused depression of MCF values of Kupffer cells. However, treatment of the diabetic male and female rats with LPS and LPS+ethanol increased the MCF values relative to those of Kupffer cells obtained from untreated diabetic rats, but administration of ethanol to diabetic rats did not have a similar effect. The induction of diabetes caused an increase in CINC generation by Kupffer cells obtained from male rats, but not from female rats. This diabetes-induced elevation of chemoattractant factor was decreased when diabetic animals were treated with LPS, ethanol, or LPS+ethanol, and the sex difference was obliterated. Thus, the induction of diabetes as well as treatment with LPS, ethanol, or LPS+ethanol in nondiabetic rats depressed the phagocytic capability of Kupffer cells, whereas the presence of endotoxemia (administration of the endotoxin LPS) or administration of LPS+ethanol reversed the diabetic effect, but ethanol intoxication did not. These findings seem to indicate a persistence of depression of host defense capacity in the ethanol-intoxicated diabetic condition. This is further reinforced by the depression of the diabetes-induced enhancement of chemotaxis when the diabetic rats became intoxicated.
我们研究了急性糖尿病和急性乙醇中毒如何调节介导免疫反应的因子，以此作为解释这两种情况下感染易感性增加的基础。我们的工作假设是，糖尿病中的乙醇中毒比单独在每种情况下观察到的更严重地损害宿主防御机制。雄性和雌性大鼠通过腹腔注射链脲佐菌素（65mg/kg）制成糖尿病模型。在注射链脲佐菌素48小时后，大鼠要么不接受治疗（对照组），要么接受以下处理：（1）乙醇（静脉推注1.75g/kg，随后以300mg/kg/h的速率进行3小时输注），（2）脂多糖[（LPS）；0.9mg/kg]，或（3）LPS+乙醇的组合。在3小时结束时，处死大鼠，通过用含胶原酶的汉克斯平衡盐溶液灌注消化肝脏以分离肝细胞和库普弗细胞。为了测量趋化因子的产生，将肝细胞（每孔2.5×10⁵个细胞）和库普弗细胞（每孔1×10⁶个细胞）培养20小时，并用上清液测量细胞因子诱导的中性粒细胞趋化因子（CINC）和活化调节的正常T细胞表达和分泌（RANTES）趋化因子。通过流式细胞术测量库普弗细胞的吞噬作用，并表示为平均通道荧光强度（MCF）。诱导糖尿病以及用LPS、乙醇或LPS+乙醇处理非糖尿病大鼠导致库普弗细胞的MCF值降低。然而，用LPS和LPS+乙醇处理糖尿病雄性和雌性大鼠相对于从未经处理的糖尿病大鼠获得的库普弗细胞，其MCF值增加，但给糖尿病大鼠施用乙醇没有类似效果。糖尿病的诱导导致从雄性大鼠而非雌性大鼠获得的库普弗细胞产生的CINC增加。当糖尿病动物用LPS、乙醇或LPS+乙醇处理时，这种糖尿病诱导的趋化因子升高降低，并且性别差异消失。因此，诱导糖尿病以及在非糖尿病大鼠中用LPS、乙醇或LPS+乙醇处理会降低库普弗细胞的吞噬能力，而内毒素血症（施用内毒素LPS）或施用LPS+乙醇可逆转糖尿病的影响，但乙醇中毒则不能。这些发现似乎表明在乙醇中毒的糖尿病状态下宿主防御能力持续降低。当糖尿病大鼠中毒时，糖尿病诱导的趋化作用增强的降低进一步强化了这一点。
