高脂饮食诱导脂肪肝小鼠骨微结构改变的研究
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国家自然科学基金(81500650);上海市科研计划项目(13ZR1432100)


Impaired bone microstructures in high fat diet induced fatty liver mice
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    摘要:

    目的:观察高脂饮食诱导的脂肪肝小鼠骨组织微结构的改变及可能机制。方法:20只4周龄C57雄性小鼠随机分成2组,标准饮食组(control,n=9)喂食标准饮食,高脂饮食组(HFD,n=11)喂食高脂食物诱导小鼠脂肪肝模型。12周实验结束后采集小鼠的血液、肝脏、股骨、胫骨样本。检测血浆丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)浓度;肝脏组织经HE和油红O染色,测定肝脏的甘油三酯(TG)及总胆固醇(TC)含量;采用micro-CT高分辨率扫描模式对两组骨样本数据进行比较,数据包括:骨总体积(TV)、骨量(BV)、骨体积分数(BV/TV)、连接密度(Conn.D)、结构模型指数(SMI)、骨小梁数目(Tb.N)、骨小梁厚度(Tb.Th)、骨小梁间距(Tb.Sp)、表观密度(Ap.Dens)、实质密度(Mat.Dens)及各向异性程度(DA),部分股骨HE染色测量骨髓脂肪含量。结果:高脂饮食组小鼠均成功建立脂肪肝模型;与标准饮食组比较,高脂饮食组小鼠的股骨具有低TV[(2.128 ± 0.591)mm3 vs.(3.570 ± 0.330)mm3,P < 0.001]、Tb.N[(1.769 ± 0.218)/mm vs.(2.284 ± 0.726)/mm,P=0.030]、SMI(2.950 ± 0.242 vs. 3.820 ± 0.729,P=0.004)以及较高的Tb.Sp[(0.595 ± 0.083)mm vs.(0.472 ± 0.116)mm,P=0.013];高脂饮食组胫骨TV[(1.127 ± 0.338)mm3 vs.(1.741 ± 0.683)mm3,P=0.017]和 SMI(2.431 ± 0.501 vs. 3.188 ± 0.465,P=0.003)也低于标准饮食组。骨髓组织形态分析后发现高脂饮食组小鼠骨髓中每mm2脂肪滴的数目(AD)显著高于标准饮食组(5.5 ± 2.2 vs. 2.6 ± 0.5,P=0.042),AD的多少与TV显著相关(股骨:r=-0.756,P=0.030;胫骨:r=-0.771,P=0.025)。结论:脂肪肝对小鼠的骨微结构有着不良影响,而骨髓的脂化可能是导致这一病理现象的潜在原因。

    Abstract:

    Objective:To evaluate the effect of fatty liver induced by high fat diet in C57 mice on bone microstructures and the possible mechanism. Methods:A total of 20 C57 male mice(4 weeks old)were enrolled and divided into two groups,fed for 12 weeks either with standard chow(control;n=9)or with high-fat diet to induce fatty liver(HFD;n=11). Blood,femur,tibia and liver samples were collected after sacrifice. Plasma alanine aminotransferase(ALT)and aspartate aminotransferase(AST)concentration were examined. The liver was stained with H&E and Oil red-O to determine if the mice were suffering from fatty liver and to measure the amount of triglyceride(TG)and total cholesterol(TC). The bone samples were scanned using a micro-CT system in a high-resolution scanning mode. Total volume(TV),bone volume(BV),total bone volume fraction(BV/TV),connectivity density(Conn.D),structural model index(SMI),trabecular number(Tb.N),trabecular thickness(Tb.Th),trabecular spacing(Tb. Sp),apparent density(Ap.Dens),material density(Mat.Dens)and degree of anisotropy(DA)were compared between control and HFD groups. The number of marrow fat drops was measured by some of the femur with HE staining. Results:In the HFD group,all mice were proved with fatty liver by liver H&E and oil red O staining. Compared to the control group,the HFD group had lower levels of TV[(2.128 ± 0.591)mm3 vs.(3.570 ± 0.330)mm3,P < 0.001)],Tb.N[(1.769 ± 0.218)/mm vs.(2.284 ± 0.726)/mm,P=0.030)],SMI(2.950 ± 0.242 vs. 3.820 ± 0.729,P=0.004)and higher levels of Tb.Sp[(0.595 ± 0.083)mm vs.(0.472 ± 0.116)mm,P=0.013]in femur. The HFD group also had lower levels of TV[(1.127 ± 0.338)mm3 vs.(1.741±0.683)mm3,P=0.017]and SMI(2.431 ± 0.501 vs. 3.188 ± 0.465,P=0.003)in tibia. Bone marrow histomorphological analysis showed that the number of adipose drop(AD)per mm2 bone marrow was significantly higher in the HFD group compared with control mice(5.5 ± 2.2 vs. 2.6 ± 0.5,P=0.042). AD was positively with TV(Femur:r= -0.756,P=0.030;Tibia:r= -0.771,P=0.025). Conclusion:Our results suggest an adverse effect of fatty liver on bone microstructure in C57 mice. Bone marrow adiposity may be a factor underlying this physiopathologic process.

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浦祥玲,崔 冉,盛 辉,曲 伸.高脂饮食诱导脂肪肝小鼠骨微结构改变的研究[J].南京医科大学学报(自然科学版),2018,(4):458-463

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  • 收稿日期:2017-11-22
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  • 在线发布日期: 2018-04-26
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