Rashid G, Luzon A A, Korzets Z, Klein O, Zeltzer E, Bernheim J
Department of Nephrology and Hypertension, Sapir Medical Center, Kfar-Saba, Israel.
Perit Dial Int. 2001 Mar-Apr;21(2):122-9.
To evaluate the effect of advanced glycation end-products (AGEs) and the inhibitor of their formation, aminoguanidine, on tumor necrosis factor-alpha (TNFalpha) production (as a functional marker) by rat peritoneal macrophages (PMphi).
Charles River rats underwent a daily intraperitoneal injection of peritoneal dialysis solution [(PDS), 4.25 g/dL dextrose; Dialine, Travenol, Ashdod, Israel] for a 2-month period (group E). Another group of rats was subjected to the same protocol with the addition of 25 mg/kg aminoguanidine (group A). Three control groups were utilized: (1) rats that were injected daily with aminoguanidine only (group AO), (2) rats that were injected with Dulbecco's phosphate-buffered saline (group D), and (3) rats in which no intervention was carried out (group C). After 2 months, PMphi were isolated from rat peritoneal effluent and their TNFalpha production measured by ELISA in cell-free culture supernatants, in both the basal state and after 24-hour stimulation with lipopolysaccharide (LPS). The concentrations of AGEs in peritoneal effluent were assayed and correlated to TNFalpha levels. PMphi obtained from normal rats were then incubated for 24 hours with (1) the peritoneal effluent of each of the above respective groups, with or without LPS; (2) increasing concentrations of AGEs (0-250 microg/mL); and (3) increasing concentrations of aminoguanidine (0-7.5 mg/mL), and TNFalpha secretion again determined.
After 2 months of daily intraperitoneal injection of PDS, in the basal state, TNFalpha production was significantly higher in PMphi isolated from the peritoneal effluent groups (groups E, A, and AO) compared to controls (group C). Following LPS stimulation, a further increase in TNFalpha secretion was seen, with a significantly greater response in group AO versus groups E, A, and D. Effluent AGEs were markedly elevated only in group E. No correlation was found between TNFalpha secretion by these PMphi and the concentration of AGEs. On incubation with the respective peritoneal effluents (groups E, A, and AO), in both the basal and stimulated state, TNFalpha production by PMphi from normal rats was significantly enhanced compared to group C. Incubation with increasing concentrations of AGEs or aminoguanidine resulted in an increase of TNFalpha secretion by these PMphi.
Following intermittent intraperitoneal administration of glucose-based PDS, rat PMphi are chronically activated, as evidenced by increased basal TNFalpha secretion. The peritoneal effluent of such treated animals is capable of stimulating TNFalpha production by normal rat PMphi. These data suggest that glucose-based PDS acts as a primer of PMphi, which retain their ability to further stimulation by LPS. Although, in vitro, AGEs promote TNFalpha secretion by normal rat PMphi, in vivo, their influence is probably modulated by other factors. Aminoguanidine has a specific inducing effect on rat PMphi, independent of glucose-based PDS.
评估晚期糖基化终末产物（AGEs）及其形成抑制剂氨基胍对大鼠腹腔巨噬细胞（PMphi）产生肿瘤坏死因子-α（TNFα）（作为一种功能标志物）的影响。
将Charles River大鼠每日腹腔注射腹膜透析液[（PDS），4.25 g/dL葡萄糖；Dialine，Travenol，以色列阿什杜德]，持续2个月（E组）。另一组大鼠接受相同方案，并添加25 mg/kg氨基胍（A组）。使用了三个对照组：（1）仅每日注射氨基胍的大鼠（AO组），（2）注射杜氏磷酸盐缓冲盐水的大鼠（D组），（3）未进行干预的大鼠（C组）。2个月后，从大鼠腹腔流出液中分离出PMphi，并通过ELISA在无细胞培养上清液中测量其在基础状态和用脂多糖（LPS）刺激24小时后的TNFα产生量。测定腹腔流出液中AGEs的浓度，并将其与TNFα水平相关联。然后将从正常大鼠获得的PMphi与（1）上述各组各自的腹腔流出液（有无LPS）孵育24小时；（2）浓度递增的AGEs（0 - 250 μg/mL）；（3）浓度递增的氨基胍（0 - 7.5 mg/mL）孵育，并再次测定TNFα分泌量。
每日腹腔注射PDS 2个月后在基础状态下，与对照组（C组）相比，从腹腔流出液组（E组、A组和AO组）分离出的PMphi中TNFα产生量显著更高。LPS刺激后，TNFα分泌进一步增加，AO组的反应明显大于E组、A组和D组。仅E组的流出液AGEs显著升高。这些PMphi的TNFα分泌与AGEs浓度之间未发现相关性。与各自的腹腔流出液（E组、A组和AO组）孵育时，在基础和刺激状态下，正常大鼠的PMphi产生的TNFα与C组相比均显著增强。与浓度递增的AGEs或氨基胍孵育导致这些PMphi的TNFα分泌增加。
间歇性腹腔给予基于葡萄糖的PDS后，大鼠PMphi被长期激活，基础TNFα分泌增加证明了这一点。此类处理动物的腹腔流出液能够刺激正常大鼠PMphi产生TNFα。这些数据表明基于葡萄糖的PDS作为PMphi的启动剂，PMphi保留了被LPS进一步刺激的能力。虽然在体外，AGEs促进正常大鼠PMphi分泌TNFα，但在体内，其影响可能受到其他因素的调节。氨基胍对大鼠PMphi有特异性诱导作用，但与基于葡萄糖的PDS无关。
