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1.天津中医药大学 中西医结合学院,天津 300193
2.天津市现代中药重点实验室,天津 300193
3.天津中医药大学 中药学院,天津 300193
[第一作者] 程丽娜,在读硕士,从事糖脂代谢研究,E-mail:chenglina1993@163.com
*康宁,博士,教授,从事糖脂代谢研究,E-mail:kangndd@163.com
收稿日期:2019-06-10,
网络出版日期:2019-09-11,
纸质出版日期:2020-02-20
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程丽娜, 曹世杰, 曲明, 等. 甘草主要成分改善L6大鼠成肌细胞胰岛素抵抗的机制[J]. 中国实验方剂学杂志, 2020,26(4):88-94.
Li-na CHENG, Shi-jie CAO, Ming QU, et al. Action Mechanism of Main Components from Glycyrrhizae Radix et Rhizoma in Improving Insulin Resistance in L6 Rat Myoblasts[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(4): 88-94.
程丽娜, 曹世杰, 曲明, 等. 甘草主要成分改善L6大鼠成肌细胞胰岛素抵抗的机制[J]. 中国实验方剂学杂志, 2020,26(4):88-94. DOI: 10.13422/j.cnki.syfjx.20200102.
Li-na CHENG, Shi-jie CAO, Ming QU, et al. Action Mechanism of Main Components from Glycyrrhizae Radix et Rhizoma in Improving Insulin Resistance in L6 Rat Myoblasts[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(4): 88-94. DOI: 10.13422/j.cnki.syfjx.20200102.
目的:
2
确定甘草主要化学成分是否通过调节L6大鼠成肌细胞内糖原合成、糖酵解途径和脂肪酸合成改善胰岛素抵抗。
方法:
2
利用0.05 mmol·L
-1
棕榈酸(PA)孵育9 h诱导L6大鼠成肌细胞建立胰岛素抵抗(IR)细胞模型;实验设正常组,模型组,甘草酸(GA,25 μmol·L
-1
)组,18
β
-甘草次酸(18
β
-GA,25 μmol·L
-1
)组,异甘草素(ILG,25 μmol·L
-1
)组和异甘草苷(ILQ,25 μmol·L
-1
)组;葡萄糖试剂盒检测细胞培养液上清葡萄糖含量;糖原检测试剂盒测定肌糖原含量;蛋白免疫印迹法(Western blot)检测固醇调节元件结合蛋白-1c(SREBP-1c),脂肪酸合成酶(FAS)和糖原合成酶激酶3
β
(GSK3
β
)的蛋白表达水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测糖酵解关键酶的mRNA水平。
结果:
2
与正常组比较,模型组IR-L6大鼠成肌细胞中葡萄糖消耗率显著下调(
P
<
0.01),细胞中的糖原含量减少(
P
<
0.05),SREBP-1c和FAS蛋白表达减少(
P
<
0.05,
P
<
0.01),磷酸果糖激酶1(PFK1),丙酮酸激酶(PK)和己糖激酶(HK) mRNA水平下调(
P
<
0.05),GSK3
β
蛋白表达增加(
P
<
0.05);与模型组比较,各给药组GA,18
β
-GA和ILG可以明显增加IR-L6大鼠成肌细胞中糖原含量(
P
<
0.05,
P
<
0.01),GA,18
β
-GA和ILQ则明显增加SREBP-1c蛋白表达(
P
<
0.05,
P
<
0.01),而GA,18
β
-GA,ILG和ILQ可明显增加FAS的蛋白表达(
P
<
0.05,
P
<
0.01)以及PFK1,PK和HK mRNA表达(
P
<
0.05,
P
<
0.01),但下调GSK3
β
蛋白表达(
P
<
0.05)。
结论:
2
甘草主要成分通过促进糖酵解和糖原合成发挥降糖作用,并且可通过调节脂肪酸合成改善胰岛素抵抗。
Objective:
2
To determine whether the main components of Glycyrrhizae Radix et Rhizoma can improve insulin resistance by regulating glycogen synthesis
glycolysis pathway and fatty acid synthesis in myoblasts of L6 rat myoblasts.
Method:
2
Insulin resistance (IR) model of L6 rat myoblasts was established through incubation with 0.05 mmol·L
-1
palmitic acid (PA) for 9 hours. Normal group
model group
glycyrrhizic acid (GA
25 μmol·L
-1
) group
18
β
-glycyrrhetinic acid (18
β
-GA
25 μmol·L
-1
) group
isoliquiritigenin (ILG
25 μmol·L
-1
) group and isoliquiritin (ILQ
25 μmol·L
-1
) group were set up
glucose content in supernatant of cell culture medium was detected by glucose kit
myoblasts glycogen content was determined by glycogen detection kit
protein expression levels of Sterol regulatory element binding protein-1c(SREBP-1c)
fatty acid synthetase (FAS) and glycogen synthase kinase3
β
(GSK3
β
) were detected by Western blot
and the mRNA expressions of key enzymes in glycolysis were detected by quantitative real-time fluorescence polymerase chain reaction(Real-time PCR).
Result:
2
Compared with those in the normal group
the glucose consumption rate was significantly down-regulated in model group (
P
<
0.01)
the glycogen content was decreased (
P
<
0.05)
the protein expressions of Sterol regulatory element binding protein-1c (SREBP-1c) and fatty acid synthase (FAS) were decreased (
P
<
0.05
P
<
0.01)
the mRNA expressions of fructose phosphate kinase 1 (PFK1)
pyruvate kinase (PK) and hexokinase (HK) were down-regulated (
P
<
0.05)
and the protein expression of glycogen synthase kinase 3 (GSK3
β
) protein was increased (
P
<
0.05). Compared with model group
GA
18
β
-GA and ILG could significantly increase glycogen content in myoblasts of IR-L6 rats (
P
<
0.05
P
<
0.01). GA
18
β
-GA and ILQ could significantly increase the expression of SREBP-1c (
P
<
0.05
P
<
0.01)
and GA
18
β
-GA
ILG and ILQ could significantly increase the expression of FAS (
P
<
0.05
P
<
0.01)
the mRNA expressions of PFK1
PK and HK (
P
<
0.05
P
<
0.01)
and down-regulate the protein expression of GSK3
β
(
P
<
0.05).
Conclusion:
2
The main components of licorice improve the insulin resistance by promoting glycolysis and glycogen synthesis and regulating fatty acid synthesis.
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