900mhz射频辐射对卵巢切除小鼠骨微结构和骨代谢的影响
首发时间:2024-03-01
摘要:[目的] 研究900mhz射频辐射对卵巢切除小鼠股骨微结构和骨代谢的影响。方法 60只3月龄c57bl/6雌性小鼠随机分为sham组(假暴露组)、ovx组(卵巢切除组)、ovx lrf组(卵巢切除后,给予50μw/cm2射频辐射)、ovx mrf组(卵巢切除后,给予150μw/cm2射频辐射)、ovx hrf组(卵巢切除后,给予450μw/cm2射频辐射)、ovx e2组(卵巢切除后,肌注0.04mg/kg雌激素处理),连续干预四周后取小鼠血清,采用elisa试剂盒检测血清e2、balp、bgp、opg、ctx-ⅰ和tracp-5b的表达水平;取双侧股骨,micro-ct检测小鼠股骨bmd以及bv、tv、bv/tv、tb.th、tb.n和tb.sp等骨微结构的变化;rt-pcr检测成骨细胞和破骨细胞分化转录因子runx2和nfatc1的表达水平,免疫组织化学染色检测股骨中opg和rankl蛋白的表达水平。结果卵巢切除后,小鼠血清e2水平显著降低,而给予雌激素处理后血清e2水平显著升高(p<0.05);小鼠血清骨形成和骨吸收结果显示,ovx组小鼠血清bgp、opg、ctx-ⅰ、tracp较假暴露组显著升高,而ovx mrf组和ovx e2组血清bgp水平显著升高,血清ctx-ⅰ和tracp-5b水平显著降低(p<0.05)。micro-ct结果显示,ovx组小鼠bmd和bv、bv/tv、tb.th、tb.n显著降低,tb.sp显著增加(p<0.05),且股骨3d图像显示骨小梁丢失严重,结构松散,而给予射频辐射和e2干预后,bmd和上述指标均有显著改善(p<0.05),骨小梁结构恶化得到明显改善,且三个剂量rf组中,以150μw/cm2 rf作用效果最为明显。rt-pcr结果显示ovx组nfatc1 和runx2 mrna表达水平显著增高(p<0.05),给予射频辐射和e2干预后,runx2 mrna表达显著增高,nfatc1 mrna显著降低(p<0.05)。免疫组化结果显示ovx组小鼠opg和rankl的表达显著增加,opg与rankl比值显著低于对照组(p<0.05);而给予射频辐射和e2干预后,opg与rankl比值显著增加,且在三个剂量rf组中,以50μw/cm2 rf作用效果最为明显。结论 150μw/cm2 900mhz射频辐射能够拮抗卵巢切除引起的小鼠血清骨代谢指标的异常、骨密度的降低、骨微结构的恶化以及骨吸收的增加。
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effect of 900mhz radiofrequency field on bone microarchitecture and metabolism in ovariectomized mice
abstract:[objective] to investigate the effect of 900mhz radiofrequency radiation on microarchitecture and metabolism of femur in ovariectomized mice. [methods]sixty 3-month-old female c57bl/6 mice were randomly divided into six groups(n=10): sham (sham exposure), ovx (ovariectomy), ovx lrf、ovx mrf、ovx hrf (50μw/cm2、150μw/cm2 and 450μw/cm2 radiofrequency radiation post ovx)、ovx e2 (0.04mg/kg estrogen injection post ovx) groups. after four weeks of continuous intervention, the mice were sacrificed. the serum samples of all mice were collected and the levels of e2, balp, bgp, opg, ctx-ⅰ and tracp-5b were detected with elisa assays. the microstructure and morphology of femur were observed with microcomputed tomography (micro-ct), the parameters of bone, including bmd, bv, tv, bv/tv, tb. n, tb.th, and tb.sp, were calculated from the reconstructed images. the mrna expression of runx2 and nfatc1, which are considered as osteoclast and osteoblast differentiation transcription factors respectively, were determined by real-time pcr. the levels of opg and rankl proteins were detected using immunohistochemical staining. [results]after ovariectomy, a significant decrease in the level of serum e2 was observed in mice, while the level of serum e2 was significantly increased following e2 treatment(p<0.05). compared with sham group, the levels of serum bgp, opg, ctx-ⅰ and tracp-5b were significantly increased in ovx group(p<0.05); following rf and e2 treatment, the levels of serum bgp in ovx mrf and ovx e2 groups were significantly increased while ctx-ⅰ and tracp-5b were significantly decreased compared that with ovx group, (p<0.05). micro- ct results showed that compared with the sham exposure group, the bmd and bv, bv/tv, tb.th, tb.n in ovx group decreased significantly while tb.sp increased significantly(p<0.05), the 3d images of femur showed severe loss of bone trabeculae and loose structure. after radiofrequency radiation and estrogen intervention, bmd and the above indicators improved significantly (p<0.05), and the deterioration of trabecular bone structure was significantly improved. among the three doses of rf groups, the effect of 150μw/cm2 rf was the most significant. compared with sham group, the expression of runx2 and nfatc1 mrna increased significantly in ovx group(p<0.05); after radiofrequency radiation and estrogen intervention, runx2 mrna expression increased significantly, while nfatc1 mrna decreased significantly (p<0.05); the western blot results showed that the levels of opg and rankl protein in ovx group were significantly increased, and the opg/rankl ratio was significantly lower than that in control group (p<0.05); compared with ovx group, opg/rankl ratio was significantly increased in ovx mrf group and ovx e2 group. [conclusion]150μw/cm2 900mhz radiofrequency irradiation could antagonize the changes of serum bone metabolism biomarkers, the decrease of bone mineral density, the deterioration of bone microstructure and the increasing bone resorption in mice induced by ovariectomy.
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900mhz射频辐射对卵巢切除小鼠骨微结构和骨代谢的影响
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