[1]李凤丽,李茂山,何　丽,等.Adropin对脂肪细胞自噬及磷脂酰肌醇-3激酶/丝氨酸苏氨酸蛋白激酶通路的影响[J].新乡医学院学报,2019,36(11):1018-1023.[doi:10.7683/xxyxyxb.2019.11.004]
　LI Feng-li,LI Mao-shan,HE Li,et al.Effects of Adropin on adipocyte autophagy and phosphatidylinositol 3 kinase/protein kinase B pathway[J].Journal of Xinxiang Medical University,2019,36(11):1018-1023.[doi:10.7683/xxyxyxb.2019.11.004]
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Adropin对脂肪细胞自噬及磷脂酰肌醇-3激酶/丝氨酸苏氨酸蛋白激酶通路的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
36
期数:
2019年11
页码:
1018-1023
栏目:
基础研究
出版日期:
2019-11-05

文章信息/Info

Title:
Effects of Adropin on adipocyte autophagy and phosphatidylinositol 3 kinase/protein kinase B pathway
作者:
李凤丽1; 李茂山2; 何　丽1; 罗　方1; 康志强1
（1.郑州大学附属郑州中心医院内分泌科，河南　郑州　450007； 2.河南省直第三人民医院骨科，河南　郑州　450006）
Author(s):
LI Feng-li1; LI Mao-shan2; HE Li1; LUO Fang1; KANG Zhi-qiang1
(1.Department of Endocrinology，Zhengzhou Central Hospital Affiliated to Zhengzhou University，Zhengzhou 450007，Henan Province，China；2.Department of Orthopaedics，the Third People′s Hospital of Henan Province，Zhengzhou 450006,Henan Province，China)
关键词:
Adropin; 脂肪细胞; 磷脂酰肌醇-3激酶/丝氨酸苏氨酸蛋白激酶通路; 自噬
Keywords:
Adropin; adipocytes; phosphatidylinositol 3 kinase/protein kinase B; autophagy
分类号:
R589.2
DOI:
10.7683/xxyxyxb.2019.11.004
文献标志码:
A
摘要:
目的　探讨Adropin对脂肪细胞自噬及磷脂酰肌醇-3激酶/丝氨酸苏氨酸蛋白激酶（PI3K/AKT）通路的影响。方法　 将小鼠胚胎成纤维前脂肪细胞3T3-L1诱导分化为成熟脂肪细胞，随机分为对照组、Adropin组、自噬激动剂组、Adropin+自噬激动剂组。对照组细胞不予特殊处理，Adropin组细胞加入Adropin（终浓度为1 000 nmol·L-1），自噬激动剂组加入西罗莫司（终浓度为100 nmol·L-1），Adropin+自噬激动剂组细胞加入Adropin（终浓度为1 000 nmol·L-1）和西罗莫司（终浓度为100 nmol·L-1），继续培养2 h；采用单丹磺酰尸胺染色检测各组细胞中自噬空泡情况，细胞免疫荧光染色法检测各组细胞微管相关蛋白轻链3（LC3）表达，采用Western blot法检测各组细胞自噬相关蛋白Beclin-1、LC3-Ⅱ、p62及PI3K/AKT通路蛋白PI3K、磷酸化PI3K（p-PI3K）、AKT、磷酸化AKT（p-AKT）的表达。结果　与对照组比较，Adropin组细胞中自噬空泡相对含量、LC3阳性细胞比例及LC3-Ⅱ、Beclin-1蛋白相对表达量显著降低（P<0.05），p62、p-PI3K、p-AKT蛋白相对表达量显著升高（P<0.05）。与对照组比较，自噬激动剂组细胞中自噬空泡相对含量、LC3阳性细胞比例及LC3-Ⅱ、Beclin-1蛋白相对表达量显著升高（P<0.05），p62、p-PI3K、p-AKT蛋白相对表达量显著降低（P<0.05）。Adropin+自噬激动剂组与对照组细胞中自噬空泡相对含量、LC3阳性细胞比例及LC3-Ⅱ、Beclin-1、p62、p-PI3K、p-AKT蛋白相对表达量比较差异无统计学意义（P>0.05）。与Adropin组比较，自噬激动剂组、Adropin+自噬激动剂组细胞中自噬空泡相对含量、LC3阳性细胞比例及LC3-Ⅱ、Beclin-1蛋白相对表达量显著升高（P<0.05），p62、p-PI3K、p-AKT蛋白相对表达量显著降低（P<0.05）。与自噬激动剂组比较，Adropin+自噬激动剂组细胞中自噬空泡相对含量、LC3阳性细胞比例及LC3-Ⅱ、Beclin-1蛋白相对表达量显著降低（P<0.05），p62、p-PI3K、p-AKT蛋白相对表达量显著升高（P<0.05）。4组细胞中PI3K、AKT蛋白相对表达量两两比较差异均无统计学意义（P>0.05）。结论　Adropin可抑制脂肪细胞自噬，其机制可能是通过激活PI3K/AKT通路而实现。
Abstract:
Objective　To investigate the effects of Adropin on adipocyte autophagy and phosphatidylinositol 3 kinase/protein kinase B (PI3K/AKT) pathway.Methods　Mouse embryonic fibroblast-adipose like cell line (3T3-L1) was induced to differentiate into mature adipocytes，then the cells were randomly divided into control group，Adropin group，autophagy agonist group and Adropin + autophagy agonist group.The cells in the control group were not given special treatment.The cells in the Adropin group were given Adropin (final concentration was 1 000 nmol·L-1).The cells in the autophagy agonist group was given sirolimus (final concentration was 100 nmol·L-1).The cells in the Adropin + autophagy agonist group were given Adropin (final concentration was 1 000 nmol·L-1) and sirolimus (final concentration was 100 nmol·L-1).Then the cells in were cultured for 2 hours.The autophagic vacuoles in cells of each group were detected by monodansylcadaverine staining.The expression of microtubule-associated protein 1 light chain 3 (LC3) in cells of each group was detected by immunofluorescence staining.The expressions of autophagy-related proteins （Beclin-1，LC3-Ⅱ，p62）and PI3K/AKT pathway-related proteins such as PI3K，phosphorylated-PI3K (p-PI3K)，AKT，phosphorylated-AKT (p-AKT) in cells of each group were detected by Western blot.Results　Compared with the control group，the relative content of autophagic vacuoles，the proportion of LC3 positive cells，the relative expressions of LC3-Ⅱ and Beclin-1 proteins was lower (P<0.05)，while the relative expressions of p62，p-PI3K and p-AKT proteins were higher in the Adropin group (P<0.05)；the relative content of autophagic vacuoles，the proportion of LC3 positive cells，the relative expressions of LC3-Ⅱ and Beclin-1 proteins in the autophagy agonist group were higher(P<0.05)，while the relative expressions of p62，p-PI3K and p-AKT proteins were lower (P<0.05).There was no significant difference in the relative content of autophagic vacuoles，the proportion of LC3 positive cells and the relative expressions of LC3-Ⅱ，Beclin-1，p62，p-PI3K and p-AKT proteins between the Adropin + autophagy agonist group and the control group (P>0.05).Compared with the Adropin group，the relative content of autophagic vacuoles，the proportion of LC3 positive cells，the relative expressions of LC3-Ⅱ and Beclin-1 proteins in the autophagy agonist group and Adropin + autophagy agonist group were higher (P<0.05)，while the relative expressions of p62，p-PI3K and p-AKT proteins were lower (P<0.05).Compared with the autophagy agonist group，the relative content of autophagic vacuoles，the proportion of LC3 positive cells，the relative expressions of LC3-Ⅱ and Beclin-1 proteins in the Adropin + autophagy agonist group were lower(P<0.05)，while the relative expressions of p62，p-PI3K and p-AKT proteins was higher (P<0.05).There was no significant difference in the relative expressions of PI3K and AKT proteins between the any two groups (P>0.05).Conclusion　Adropin can inhibit autophagy of adipocytes，and its mechanism may be involve in activating PI3K/AKT pathway.

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更新日期/Last Update: 2019-11-05