外源性硫化氢对去卵巢骨质疏松大鼠骨代谢及护骨素/细胞核因子κB受体活化因子配体表达的影响

摘要

目的：探讨外源性硫化氢(H2S)对去卵巢骨质疏松(OP)大鼠骨代谢及护骨素(OPG)和细胞核因子资B受体活化因子配体(RANKL)表达的影响。方法将50只健康SD雌性大鼠随机分为对照组、去卵巢OP模型组和外源性H2S供体硫氢化钠(NaHS)(10、50、100μmol/kg)组，每组10只。12周后检测各组大鼠的尿钙和尿脱氧吡啶啉(Dpd)及血清钙、磷、碱性磷酸酶(ALP)和雌二醇(E2)水平，采用双能X线吸收法测量大鼠右侧股骨的骨密度(BMD)，采用Western blot检测股骨中OPG和RANKL的表达。结果与对照组比较，去卵巢OP模型组大鼠血清E2[(76.83±8.12)pmol/L比(11.05±1.42)pmol/L，t=5.74，P<0.05]和BMD水平[(0.25±0.03)g/cm2比(0.14±0.01)g/cm2，t=3.85，P<0.05]显著降低，尿钙[(0.85±0.08)mg/L比(1.88±0.21)mg/L，t=4.65，P<0.05]、尿Dpd[(14.67±1.28)nmol/mmol比(22.34±1.75)nmol/mmol，t=3.81，P<0.05]、血清钙[(3.29±0.27)mmol/L比(4.68±0.52)mmol/L，t=3.98，P<0.05]、血清磷[(2.49±0.35)mmol/L比(4.03±0.59)mmol/L，t=4.31，P<0.05]和ALP水平[(78.65±5.23)U/L比(167.35±1.75)U/L，t=4.47，P<0.05]均显著升高；OPG的表达显著降低[(0.68±0.09)比(0.27±0.04)，t=3.82，P<0.05]，而RANKL的表达显著增加[(0.18±0.02)比(0.66±0.09)，t=3.91，P<0.05]。与去卵巢OP模型组比较，NaHS(50、100μmol/kg)组大鼠股骨的BMD显著增加[(0.14±0.01)g/cm2比(0.19±0.02)g/cm2和(0.23±0.03)g/cm2，F=6.42，P<0.05]，尿钙[(1.88±0.21)mg/L 比(1.39±0.09)mg/L 和(0.97±0.09)mg/L，F=4.67，P <0.05]、尿 Dpd [(22.34±1.75)nmol/mmol 比(18.43±2.04)nmol/mmol和(15.75±1.14)nmol/mmol，F=3.92，P<0.05]、血清钙[(4.68±0.52)mmol/L比(3.98±0.47)mmol/L和(3.56±0.25)mmol/L，F=3.89，P<0.05]、血清磷[(4.03±0.59)mmol/L比(3.11±0.42)mmol/L和(2.68±0.26)mmol/L，F=4.23，P<0.05]和ALP水平[(167.35±1.75)U/L比(115.69±8.72)U/L和(87.61±9.56)U/L，F=4.15，P<0.05]显著降低，OPG的表达显著增加[(0.27±0.04)比(0.48±0.05)和(0.64±0.08)，F=4.84，P<0.05]，而RANKL的表达显著降低[(0.66±0.09)比(0.37±0.05)和(0.22±0.04)，F=5.16，P<0.05]。结论外源性H2S抑制了卵巢切除所引起的OP，其作用机制可能与H2S上调骨组织中OPG的表达而降低RANKL的表达有关。%Objective To investigate the effect of extrogenous hydrogen sulfide (H2S) on the bone metabolism and osteo-protegerin/receptor activator of nuclear factor kappa Bligand(OPG/RANKL) expressions in ovariectomized rats with osteoporosis. Methods 50 SD female rats were randomly divided into control group, ovariectomized osteoporosis model group, and extrogenous H2S donator sodium hydrosulfide (NaHS) (10, 50, 100 μmol/kg) treatment group, with 10 rats in each group. After 12 weeks, the levels of urine calcium and deoxypyridinoline (Dpd), serum calcium, serum phosphorus, serum alkaline phosphatase (ALP) and estradiol (E2) were measured. Bone mineral density (BMD) was measured by a dual-energy X-ray absorptiometry. The expessions of OPG and RANKL in femur were measured by Western blot. Re-sults Compared with the sham control group, the levels of E2 in ovariectomized osteoporosis model group [(76.83±8.12) pmol/L vs (11.05±1.42) pmol/L, t=5.74, P<0.05] and BMD [(0.25±0.03) g/cm2 vs (0.14±0.01) g/cm2, t=3.85, P<0.05] were significantly decreased, the levels of urine calcium [(0.85±0.08) mg/L vs (1.88±0.21) mg/L, t=4.65, P<0.05] and Dpd [(14.67±1.28) nmol/mmol vs(22.34±1.75) nmol/mmol,t=3.81,P< 0.05], serum calcium [(0.85±0.08) mg/L vs(1.88±0.21) mg/L, t=4.65, P< 0.05], serum phosphorus [(2.49±0.35) mmol/L vs (4.03±0.59) mmol/L, t=4.31, P< 0.05] and serum ALP [(78.65±5.23) U/L vs(167.35±1.75) U/L, t=4.47, P< 0.05] were significantly increased, the expression of OPG in the ovariectomized osteoporosis model group was significantly decreased [(0.68±0.09) vs (0.27±0.04), t=3.82, P < 0.05] and the expression of RANKL in the ovariectomized osteoporosis model group was significantly increased [(0.18±0.02) vs (0.66±0.09), t=3.91, P< 0.05] in the ovariectomized osteoporosis model group. Compared with the o-variectomized osteoporosis model group, the level of BMD in NaHS group (50, 100 μmol/kg) [(0.14±0.01) g/cm2 vs(0.19±0.02) g/cm2 and (0.23±0.03) g/cm2,F=6.42,P < 0.05] were significantly increased, the levels of urine calcium [(1.88±0.21) mg/L vs (1.39±0.09) mg/L and (0.97±0.09) mg/L, F=4.67, P< 0.05] and Dpd [(22.34±1.75) nmol/mmol vs (18.43±2.04) nmol/mmol and (15.75±1.14) nmol/mmol, F=3.92, P<0.05], serum calcium [(4.68±0.52) mmol/L vs (3.98±0.47) mmol/L and (3.56±0.25) mmol/L, F=3.89, P<0.05], serum phosphorus [(4.03±0.59) mmol/L vs (3.11±0.42) mmol/L and (2.68±0.26) mmol/L, F=4.23, P<0.05] and serum ALP [(167.35±1.75) U/L vs (115.69±8.72) U/L and (87.61±9.56) U/L, F=4.15, P< 0.05] were significantly decreased, the expression of OPG in the ovariectomized osteoporosis model group were significantly up-regulated [(0.27±0.04) vs (0.48±0.05) and (0.64±0.08), F=4.84, P< 0.05] and the expression of RANKL were significantly down-regulated [(0.66±0.09) vs(0.37±0.05) and (0.22±0.04), F=5.16, P<0.05] in the o-variectomized osteoporosis model group. Conclusion Extrogenous H2S inhibits the osteoporosis induced by ovariotomy in rats, the mechanism may be related to the up-regulation of OPG expression and the down-regulation of RANKL ex-pression induced by H2S.