莱菔硫烷对高脂诱导肥胖大鼠肾脏线粒体复合体氧化损伤的保护作用

Protective effects of sulforaphane on the oxidative damage of kidney mitochondria complex in obese rats induced by high-fat diet

目的：研究高脂膳食对大鼠肾脏线粒体氧化损伤情况，并分析莱菔硫烷对高脂饮食诱导的大鼠肾脏线粒体氧化损伤的保护作用。方法将88只8周龄（体重180～200 g）雄性SPF级SD大鼠适应性喂养1周后，采用随机数字法选取8只作为基础对照组，并仅喂养基础饲料至实验结束。其余大鼠喂养高脂饲料2周后，选取体重增量≥40％的大鼠诱导肥胖模型，共32只。采用随机数字表法将肥胖模型大鼠平均分为高脂对照组，莱菔硫烷低、中、高剂量组。莱菔硫烷低、中、高剂量组灌胃的莱菔硫烷剂量分别为5、10、20 mg／kg，同时喂养高脂饲料。喂养5周后，取大鼠肾脏，检测线粒体氧化损伤的相关指标，并分析不同组间指标变化。结果高脂对照组大鼠的肾脏活性氧簇（0.26±0.04）和丙二醛含量[（0.87±0.05）U／mg ]均高于基础对照组[（0.20±0.02）、（0.57±0.08）U／mg]（t值分别为－3.02、－4.72，P值均＜0.05），总抗氧化能力（T-AOC）活性[（0.43±0.04） U／mg]、线粒体膜电位（MMP）水平（12.09±1.56）均低于基础对照组[（0.48±0.04） U／mg、16.08±3.12]（t值分别为2.06、2.28，P值均＜0.05）。灌胃干预后，莱菔硫烷低、中、高剂量组丙二醛含量[（0.67±0.05）、（0.55±0.05）、（0.56±0.07）U／mg]均低于高脂对照组[（0.87±0.05） U／mg]（t值分别为3.65、5.71、5.60，P值均＜0.05），T-AOC活性[（0.49±0.05）、（0.55±0.05）、（0.54±0.04） U／mg]、总超氧化物歧化酶（T-SOD）活性[（61.07±2.79）、（55.95±2.39）、（60.26±6.02） U／mg]、MMP水平（17.17±2.52、18.24±2.54、18.21±3.65）均高于高脂对照组[（0.43±0.04） U／mg、（47.22±2.43） U／mg、12.09±1.56]（T-AOC活性t值分别为－2.36、－4.83、－4.30，T-SOD活性t值分别为－6.37、－4.71、－5.99，MMP水平t值分别为－2.90、－3.52、－3.50，P值均＜0.05）；莱菔硫烷低、中剂量组谷胱甘肽过氧化物酶（GSH-Px）活性[（69.12±8.63）、（64.43±6.58）U／mg]均高于高脂对照组[（53.03±5.70）U／mg]（t 值分别为－3.82、－2.71，P 值均＜0.05），高剂量组 GSH-Px 活性[（60.02±7.05）U／mg]与高脂对照组差异无统计学意义（t＝－1.66，P＞0.05）。结论高脂膳食可以诱导肾脏线粒体出现氧化损伤。莱菔硫烷对高脂饮食诱导肥胖大鼠肾脏线粒体氧化损伤及线粒体复合体活性的降低具有一定的保护作用。

Objective To realize the oxidative damage of kidney mitochondrial complex in obese rats induced by high-fat diet and investigate the protective effects of sulforaphane against the damage . Methods Eighty-eight adult male SD rats were used , after 1 week adaptability feeding , 8 rats were selected as control group and given low-fat diet.The other 80 rats were given high-fat diet.After 2 weeks, the 32 diet-induced obesity models were choosen whose weight gain was higher than 40%.The 32 rats were randomly divided into 4 groups, i.e.high fat group, high fat ＋sulforaphane low dose group , high fat ＋sulforaphane middle dose group and high fat ＋sulforaphane high dose group.The rats in the sulforaphane＂nbsp;low, middle and high dose groups were orally administered with sulforaphane 5, 10 and 20 mg/kg, all the 4 groups were kept feeding high-fat diet for 5 weeks.All rats were sacrificed and their kidneys were removed to assay the index of oxidative damages .Results The content of ROS （0.26 ±0.04） and MDA（（0.87 ± 0.05） U/mg） in the hight-fat group were significantly higher than those in the control group （（0.20 ± 0.02）,（0.57 ±0.08） U/mg） （t values were -3.02 and -4.72, P 〈0.05）.The activity of T-AOC （（0.43 ±0.04） U/mg） and MMP （12.09 ±1.56） were lower than the control group （（0.48 ± 0.04 U/mg,（16.08 ±3.12） ） （t values were 2.06 and 2.28,P 〈0.05）.Gavage intervention with sulforaphane , the MDA amount （ （ 0.67 ±0.05 ） , （ 0.55 ±0.05 ） , （ 0.56 ±0.07 ） U/mg ） in the sulforaphane low , middle and high dose groups were lower than the hight-fat group （ （ 0.87 ±0.05 ） U/mg （t values were 3.65,5.71 and 5.60.P〈0.05）.The activity of T-AOC（（0.49 ±0.05）,（0.55 ±0.05）, （0.54 ±0.04） U/mg）, T-SOD（（61.07 ±2.79）, （55.95 ±2.39）, （60.26 ±6.02） U/mg） and the level of MMP（（17.17 ±2.52）, （18.24 ±2.54）, （18.21 ±3.65）） were higher than in the high-fat group （（0.43 ±0.04） U/mg,（47.22 ±2.43） U/mg,（12.09 ±1.56））（tT-AOC values were -2.36, -4.83 and-4.30;tT-SOD values were -6.37, -4.71 and -5.99; tMMP values were -2.90,-3.52 and -3.50, P〈0.05 ）.The activity of GSH-Px in the sulforaphane low and middle dose groups （ （ 69.12 ±8.63 ） , （64.43 ±6.58） U/mg） were higher than those in the high-fat group（（53.03 ±5.70） U/mg）（t values were -3.82 and -2.71,P〈0.05）.But there were no significant difference between the high dose group （（60.02 ±7.05） U/mg） and the high-fat group （t=-1.66,P〉0.05）.Conclusion High-fat diet can induce the mitochondrial oxidative dysfunction in kidney , and sulforaphane shows protective effect on the kidney mitochondrial complex from oxidative damage in obese rats induced by high -fat diet.