低氧下运动训练抑制载脂蛋白E敲除小鼠脂肪组织细胞焦亡的研究

关键词：载脂蛋白E  脂肪组织  细胞焦亡  低氧  运动

Keywords:ApoE  adipose tissue  pyroptosis  hypoxia  exercise

摘要：

      摘要 目的：探究6周运动训练对慢性间歇低氧暴露下高脂膳食载脂蛋白E敲除（ApoE-/-）小鼠脂肪组织细胞焦亡的影响。 方法：40雄性ApoE-/-小鼠随机分为常氧不运动（N）、低氧不运动（H）、常氧运动（E）、低氧运动（HE）4组（高脂饲料），10只C57BL/6J小鼠为野生对照（W，普通饲料）。H组进行间歇低氧暴露，E组进行跑台运动，HE组在低氧环境下进行同E组运动。干预6周后，检测体质量、血脂；HE染色分析脂肪细胞横截面积；TUNEL染色评估凋亡；免疫荧光和Western Blot检测焦亡蛋白（NLRP3， ASC， NF-κB， Caspase-1， GSDMD）；PCR芯片筛选差异基因并用qPCR验证。 结果：①各组体质量无差异；HE组HDL-C显著高于H组（P<0.05）。②N组脂肪细胞横截面积显著小于W组，H组显著小于N组，HE组显著大于H组（P<0.05）。③H组细胞凋亡水平和NLRP3积分光密度（IOD）值显著高于N组，E组显著低于N组；HE组ACS和NLRP3的IOD值低于H组（P<0.05）。④H组NF-κB蛋白含量和 Caspase1/pro-Caspase1比值显著高于N组；H组NF-κB蛋白含量和 Caspase1/pro-Caspase1比值显著高于N组，HE组NLRP3、NF-κB蛋白含量、Caspase1/pro-Caspase1和cleaved-GSDMD/pro-GSDMD比值显著低于H组（P<0.05）。⑤与N组比，H组焦亡关键基因Nlrp3表达上调，而HE组差异表达基因数目减少且关键基因表达受抑（qPCR验证一致）。 结论：6周运动训练可抑制慢性间歇低氧暴露诱导的ApoE-/-小鼠脂肪组织细胞焦亡，改善血脂异常和脂肪细胞形态。

Abstract:

      Abstract Objective： To investigate the effect of 6-week exercise training on pyroptosis of adipocytes in apolipoprotein E knockout （ApoE-/-） mice fed a high-fat diet under chronic intermittent hypoxia exposure. Method： Forty male ApoE-/- mice were randomly assigned to four groups（fed high-fat diet）： normoxia sedentary（N），hypoxia sedentary（H），normoxia exercise（E），and hypoxia exercise（HE）. Ten C57BL/6J mice served as wild-type controls（W，fed standard diet）. Group H underwent intermittent hypoxia exposure. Group E performed treadmill exercise. Group HE performed the same treadmill exercise as group E but under hypoxic conditions. After 6 weeks of intervention， body weight and blood lipids were measured. Hematoxylin and eosin （HE） staining was used to analyze adipocyte cross-sectional area. TUNEL staining was used to assess apoptosis. Immunofluorescence and Western Blot were used to detect pyroptosis-related proteins（NLRP3，ASC，NF-κB，Caspase-1，GSDMD）. PCR arrays were used to screen for differentially expressed genes， which were validated by qPCR. Result： ①No significant differences in body weight were observed among groups. HDL-C level in group HE was significantly higher than that in group H （P<0.05）. ②The adipocyte cross-sectional area was significantly smaller in group N than in group W， significantly smaller in group H than in group N， and significantly larger in group HE than in group H （P<0.05）. ③The cell apoptosis levels and NLRP3 integrated optical density （IOD） values in group H were significantly higher than in group N， while they were significantly lower in group E than in group N. ASC and NLRP3 IOD values in group HE were lower than those in group H （P<0.05）. ④The NF-κB protein content and the Caspase1/pro-Caspase1 ratio in group H were significantly higher than those in group N. Furthermore， in group HE， the levels of NLRP3， NF-κB protein content， and the ratios of Caspase1/pro-Caspase1 and cleaved-GSDMD/pro-GSDMD were significantly lower than those in group H （P<0.05）. ⑤Compared to group N， the expression of the key pyroptosis gene Nlrp3 was upregulated in group H. In contrast， group HE showed a reduced number of differentially expressed genes and suppressed expression of key pyroptosis genes （confirmed by qPCR）. Conclusion： Six weeks of exercise training can suppress chronic intermittent hypoxia-induced adipose tissue pyroptosis in ApoE-/- mice， while also ameliorating dyslipidemia and improving adipocyte morphology.