卵巢切除对高胰岛素血症MKR小鼠糖脂代谢的影响

doi:10.13481/j.1671-587X.20220517

摘要/Abstract

摘要：

目的探讨正常或高胰岛素血症情况下卵巢切除对小鼠血糖、脂肪组织含量和肝脏脂质沉积的影响。方法 6周龄雌性骨骼肌特异性胰岛素样生长因子1受体功能缺失（MKR）高胰岛素血症小鼠20只随机分为MKR假手术组（MKR组）和MKR卵巢切除组（MKR OVX组），每组10只；野生型（WT）FVB/N小鼠20只随机分为WT假手术组（WT组）和WT卵巢切除组（WT OVX组），每组10只。术后恢复饲养2周，监测各组小鼠体质量和血糖水平；葡萄糖耐量试验（GTT）和胰岛素耐量试验（ITT）检测小鼠葡萄糖耐受能力和胰岛素敏感性；术后12周处死小鼠，称量各组小鼠肝脏和脂肪组织质量，HE染色、糖原染色和油红O染色观察各组小鼠肝脏和性腺脂肪组织病理形态表现，实时荧光定量PCR（RT-qPCR）法检测各组小鼠性腺脂肪组织中脂代谢相关基因mRNA表达水平，Western blotting法检测各组小鼠性腺脂肪组织中激素敏感脂肪酶（HSL）蛋白表达水平。结果与WT组比较，MKR组小鼠体质量明显降低（P<0.05）；与MKR组比较，MKR OVX组小鼠体质量明显增加（P<0.05），但与WT组和WT OVX组比较，MKR OVX组小鼠体质量降低（P<0.05）。MKR OVX组小鼠血糖逐渐升高，出现严重葡萄糖不耐受和胰岛素抵抗。与WT组比较，WT OVX组小鼠肝脏出现脂肪变性，糖原减少，MKR OVX组小鼠肝脂肪变性程度增加且糖原减少。与WT组比较，WT OVX组小鼠性腺脂肪和肠系膜脂肪系数明显升高（P<0.05）；与MKR组比较，MKR OVX组小鼠皮下脂肪、性腺脂肪和肾周脂肪系数明显升高（P<0.05）。HE染色，与WT组比较，MKR组小鼠脂肪细胞明显缩小，WT OVX组和MKR OVX组小鼠脂肪细胞均增大，但与WT组和WT OVX组比较， MKR OVX组小鼠脂肪细胞仍缩小。RT-qPCR和Western blotting法检测，与WT组比较，MKR组小鼠性腺脂肪组织中HSL mRNA和蛋白表达水平升高（P<0.05）；与MKR组比较，MKR OVX组小鼠性腺脂肪组织中HSL mRNA和蛋白表达水平降低（P<0.05）。结论卵巢切除会导致小鼠糖代谢紊乱、脂肪组织质量增加和肝脏脂质沉积，而高内源性胰岛素水平和胰岛素抵抗会进一步加重因卵巢切除导致的糖脂代谢紊乱。

关键词: 高胰岛素血症, 卵巢切除, 骨骼肌特异性胰岛素样生长因子1受体功能缺失小鼠, 糖脂代谢

Abstract:

Objective To investigate the effects of ovariectomy on the blood glucose， adipose tissue content and liver lipid deposition in the normal or hyperinsulinemia mice. Methods Twenty 6-week-old female loss of skeletal muscle-specific insulin-like growth factor-1 receptor function（MKR） hyperinsulinemia mice were randomly divided into MKR sham operation group （MKR group） and MKR ovariectomy group （MKR OVX group），with 10 mice in each group；twenty wild-type （WT） FVB/N mice were randomly divided into WT sham operation group（WT group） and WT ovariectomy group（WT OVX group），with 10 mice in each group. After two weeks of recovery feeding， the body weights and blood glucose levels of the mice in various groups were monitored. Glucose tolerance abilities and insulin sensitivities of the mice in various groups were detected by glucose tolerance test（GTT） and insulin tolerance test（ITT）. The mice were sacrificed 12 weeks after surgery，and the weights of liver and adipose of the mice in various groups were weighed.The pathomorphology of liver and gonadal adipose tissue of the mice in various groups were observed by HE staining，glycogen staining and oil red O staining. The expression levels of lipid metabolization-related gene mRNA in gonadal adipose tissue of the mice in various groups were detected by real-time fluorescence quantitative PCR（RT-qPCR） method. Western blotting method was used to detect the expression levels of hormone sensitive lipase（HSL） in gonadal adipose tissue of the mice in various groups. Results The body weight of the mice in MKR group was significantly decreased compared with WT group （P<0.05）； compared with MKR group， the body weight of the mice in MKR OVX group was significantly increased （P<0.05）， but compared with WT and WT OVX groups，the body weight of the mice in MKR OVX group was decreased （P<0.05）. In MKR OVX group， the blood glucose of the mice was gradually increased， and severe glucose intolerance and insulin resistance appeared. Compared with WT group， liver steatosis was found and glycogen was decreased in WT OVX group （P<0.05）， while the liver steatosis degree was increased and glycogen was decreased significantly in MKR OVX group.Compared with WT group，the gonadal adipose index and mesenteric adipose index of the mice in WT OVX group were increased（P<0.05）；compared with MKR group，the gonadal adipose index and perirenal adipose index of the mice in MKR OVX group was increased（P<0.05）.The HE staining results showed that the adipocytes in MKR group were significantly shunk compared with WT group， and the adipocytes were enlarged in WT OVX group and MKR OVX group，the adipocytes in MKR OVX group were still shunk compared with WT group and WT OVX group. Compared with WT group， the expression levels of HSL mRNA and protein in gonadal adipose tissue of the mice in MKR group were increased （P<0.05）， while the expression levels of HSL mRNA and protein in gonadal adipose tissue of the mice in WT OVX group and MKR OVX group were decreased（P<0.05）. Conclusion Ovariectomy can lead to dysglycemia， adipose tissue mass increasing and lipid deposition in the liver in the mice. High endogenous insulin levels and insulin resistance can further aggravate the dysglycemia caused by ovariectomy.

Key words: Hyperinsulinemia, Ovariectomy, Loss of skeletal muscle-specific insulin-like growth factor-1 receptor function mice, Glucose and lipid metabolism

中图分类号:

R587.3

姚庆,王丹,王小双,吴英杰,冉丽媛. 卵巢切除对高胰岛素血症MKR小鼠糖脂代谢的影响[J]. 吉林大学学报(医学版), 2022, 48(5): 1229-1237.

Qing YAO,Dan WANG,Xiaoshuang WANG,Yingjie WU,Liyuan RAN. Effect of ovariectomy on glucose and lipid metabolism in hyperinsulinemia MKR mice[J]. Journal of Jilin University(Medicine Edition), 2022, 48(5): 1229-1237.

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Group	Liver weight(m/g)	Liver index	Brown adipose index	Subcutaneous adipose index	Gonadal adipose index	Perirenal adipose index	Mesenteric adipose index
WT	1.499±0.197	5.236±0.955	0.491±0.059	2.444±0.908	3.996±0.841	1.776±0.995	0.509±2.223
WT OVX	1.555±0.074	5.225±0.447	0.553±0.042	2.677±0.579	4.872±1.013^*	2.092±0.628	1.785±0.288^*
MKR	1.445±0.243	6.395±0.571^*△	0.466±0.052^△	1.354±0.284^*△	1.332±0.374^*△	0.399±0.067^*△	1.083±0.076^*△
MKR OVX	1.561±0.027	6.645±0.396^*△	0.417±0.018^△	1.748±0.335^*△#	2.583±0.404^*△#	0.930±0.203^*△#	1.109±0.066^*△

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