羟乙膦酸钠和1,25(OH)_2D_3对去卵巢大鼠IL1、IL6和TNFα分泌的影响

EFFECT OF INTERMITTENT ETIDRONATE DISODIUM AND 1,25(OH)2D3 ON SECRETION OF IL1, IL6 AND TNFα BY SPLEEN MACROPHAGES IN OVARIECTOMIZED RATS

５０只雌性Ｗｉｓｔａｒ大鼠，１０只行假性去卵巢术，４０只切除双侧卵巢。去卵巢大鼠再分为单纯去卵巢组、周期口服羟乙膦酸钠组、周期口服１，２５（ＯＨ）２Ｄ３组、周期序贯口服羟乙膦酸钠和１，２５（ＯＨ）２Ｄ３组。去卵巢第２６周末处死后取脾脏制备巨噬细胞，用ＭＴＴ法检测其在细菌脂多糖刺激下白细胞介素１（ＩＬ１）、白细胞介素６（ＩＬ６）和肿瘤坏死因子α（ＴＮＦα）分泌水平。结果表明，单纯去卵巢组的ＩＬ１分泌水平显著增高（Ｐ＜０．０５）；周期口服羟乙膦酸钠组在停药６周后，ＩＬ１和ＩＬ６分泌水平显著增高（Ｐ＜０．０５）；周期序贯口服羟乙膦酸钠和１，２５（ＯＨ）２Ｄ３组的ＩＬ１、ＩＬ６和ＴＮＦα分泌水平较单纯去卵巢组显著降低，近似于假性去卵巢组，亦低于单纯用羟乙膦酸钠或１，２５（ＯＨ）２Ｄ３组。提示羟乙膦酸钠和１，２５（ＯＨ）２Ｄ３联合序贯应用可使脾脏巨噬细胞ＩＬ－１、ＩＬ－６和ＴＮＦα分泌水平接近正常大鼠分泌水平，从而有可能抑制因雌激素减少所诱发的破骨细胞生成过多和活性增强，减少骨丢失。

Estrogen deficiency has been known to increase bone resorption, which is regulated by various cytokines such as IL1, IL6 and TNFα, secreted by cell of monocyte_macrophage series, through stimulating osteoclastogenesis and osteoclast activation. The effect of etidronate disodium (HEBP) and 1,25(OH)2D3 on secretion of IL_1, IL_6 and TNFα by spleen macrophages in ovariectomized rats was investigated.Forty female Wistar rats, 8 weeks of age, were subjected to ovariectomy (OVX) while 10 to sham_surgery (sham_OVX). 8 weeks later, thise 50 rats were divided into 5 groups: (1)sham_OVX, (2)OVX, (3)OVX+HEBP (medium phosphonate for 1 week, HEBP 30mg/kg BW/day orally for 2 weeks followed by 6 weeks off treatment), (4)OVX+1,25(OH)2D3 (non_treatmentfor3weeksfollowedby 6 weeks of oral 1,25(OH)2D3 0.4μg/kg BW/day), (5)OVX+HEBP+1,25(OH)2D3 (medium phosphonate for 1 weeks, HEBP for 2 weeks followed by 1,25(OH)2D3 for 6 weeks). After 2 cycles of treatment (18 weeks) in 4 OVX groups, all five groups of rats were sacrified at end of 26 weeks postsurgery. IL1, IL6 and TNFα activities released by lipopolysaccharide_stimulated spleen macrophages were determined by MTT method. The results showed that IL1 secretion in OVX rats of group 2 was significantly higher than that in sham_OVX rats (P<0.05). WithintermittentHEBPtreatment, secretion of IL1 and IL6 in group 3 was significantly higher than that of other groups (P<0.01 or P<0.05). But since the result was determined after 6 weeks of cessation of treatment it indicated that the effect of HEBP could not last 6 weeks, though being a powerful inhibitor of bone resorption. Comparing with sham_OVX rats, IL1 secretion with intermittent 1,25(OH)2D3 in group 4 was, to a certain extent, decreased. With coherent treatment of HEBP and 1,25(OH)2D3 in group 5, secretion of IL1, IL6 and TNFα was nearly similar to that of sham_OVX rats, but was significantly lower than that of OVX rats and also lower than those of OVX+HEBP and OVX+1,25(OH)2D3 rats. These results suggest that coherent therapy with HEBP and 1,25(OH)2D3 , especially with the effect of 1,25(OH)2D3 for longterm treatment, may be more effective to decrease the IL1, IL6 and TNFα secretion by spleen macrophages, thus to suppress the osteoclastogenesis and osteoclast activation induced by estrogen deficiency.