Astragaloside IV ameliorates insulin induced insulin resistance in HepG2 cells through reactive oxygen species mediated c-Jun N-terminal kinase pathway

doi:10.19852/j.cnki.jtcm.2023.01.007

Journal of Traditional Chinese Medicine ›› 2023, Vol. 43 ›› Issue (1): 60-67.DOI: 10.19852/j.cnki.jtcm.2023.01.007

• Original articles • Previous Articles Next Articles

Astragaloside IV ameliorates insulin induced insulin resistance in HepG2 cells through reactive oxygen species mediated c-Jun N-terminal kinase pathway

YE Xiaomei¹^,², XING Xiaowei¹, YUAN Kangrui¹, WANG Dongming⁴, WU Dudu¹, CHEN Zhi¹(), YU Zhiqiang³()

1 School of Pharmacy, Guangdong Medical University, Dongguan 523808, China
2 Department of Clinical Pharmacy, SSL Central Hospital of Dongguan City (the Third People's Hospital of Dongguan City), Dongguan 523326, China
3 School of Pharmaceutical Sciences, Guangdong Provincial Key Laboratory of New Drug Screening, Southern Medical University, Guangzhou 510515, China
4 Department of Pharmacy, Central People's Hospital of Zhanjiang, Zhanjiang 524037, China

Received:2021-03-22 Accepted:2021-06-12 Online:2023-02-15 Published:2023-01-10
Contact: CHEN Zhi,YU Zhiqiang
About author:YU Zhiqiang, School of Pharmaceutical Sciences, Guangdong Provincial Key Laboratory of New Drug Screening, Southern Medical University, Guangzhou 510515, China. zhiqiang.yu.1@asu.edu. Telephone:+86-769-22896321; +86-769-22896560
CHEN Zhi, School of Pharmaceutical Sciences, Guangdong Provincial Key Laboratory of New Drug Screening, Southern Medical University, Guangzhou 510515, China. czwududu@126.com;
Supported by:
National Natural Science Foundation of Guangdong Province(2017A030310666);National Natural Science Foundation of Guangdong Province(2018A030307003);Guangdong Medical University Nanhai Marine Biomedical Resources R&D Public Service Platform Open Fund Project(2HC18013);Guangdong Medical University Nanhai Marine Biomedical Resources R&D Public Service Platform Open Fund Project(2HC18016);“Group-type” Special Support Project for Education Talents in Universities(4SG19045G);Undergraduate Science & Technology Innovation Foundation of Guangdong Province(201810571046);Undergraduate Science & Technology Innovation Foundation of Guangdong Province(201810571073);Medical Science and Technology Development Foundation of Guangdong Province(A2016355);The Opening Project of State Key Laboratory of Polymer Materials Engineering (Sichuan University)(sklpme2018-4-23)

Abstract

Abstract:

OBJECTIVE: To investigate the effects and elucidate the mechanism of Astragaloside IV (AS-IV) for insulin resistance (IR) and type 2 diabet es mellitus (T2DM).
METHODS: CCK8 kit was used to detect cell viability, glucose detection kit was used to detect the concentration of glucose in cell supernatant, reactive oxygen species (ROS) detection kit and Western blot were used to explore the mechanism of Astragaloside IV (AS-IV) in improving IR. A diabetic rat model was also established by feeding high sugar and fat diet and streptozotocin (STZ) injection. After treatment with AS-IV, rosiglitazone (ROZ), or normal saline, the fasting blood glucose (FBG), C peptide (C-P), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and the glucose tolerance were assessed.
RESULTS: AS-IV could effectively reduce the content of ROS and increase the glucose uptake in high insulin-treated IR-type HepG2 cells. The results of molecular mechanisms indicated that AS-IV could improve insulin resistance by reducing JNK phosphorylation and regulating c-Jun N-terminal kinase (JNK) downstream protein expression. Additionally, AS-IV could significantly reduce the levels of FBG, TNF-α, IL-6 and the glucose tolerance in diabetic rats (P < 0.05 or < 0.01). The high and medium dose groups of AS-IV could significantly increase the C-P levels in diabetic rats (P < 0.05 or < 0.01).
CONCLUSIONS: Our results indicated that AS-IV improve liver IR through the JNK pathway and ROS, which meant a new molecular target for the treatment of diabetes. The AS-IV also helped to prevent and improved the insulin resistance of rats.

Key words: Astragaloside IV, Insulin resistance, Glucose consumption, ROS, JNK, HepG2 cells

YE Xiaomei, XING Xiaowei, YUAN Kangrui, WANG Dongming, WU Dudu, CHEN Zhi, YU Zhiqiang. Astragaloside IV ameliorates insulin induced insulin resistance in HepG2 cells through reactive oxygen species mediated c-Jun N-terminal kinase pathway[J]. Journal of Traditional Chinese Medicine, 2023, 43(1): 60-67.

Figures/Tables 6

Figure 1 Effects of AS-IV on the viability of HepG2 cells A-D: cells were treated with 0.1, 1, 5, 10, 100 or 150 μM AS-IV for 24 (A), 48 (B), 72 (C) and 96h (D), and inhibition of cell viability was determined by MTT assay. Results are expressed as the mean ± standard deviation of three independent experiments (n = 3). aP < 0.05 compared with untreated cells. MTT: 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide.

Figure 2 Effects of AS-IV on ameliorating glucose consumption in insulin resistance N: control group, given conventional feed without other interfering factors. M: model group, given high sugar and fat feed but no other interfering factors; ROZ: positive group (30 μM rosiglitazone). [Results are expressed as the mean ± standard deviation of three independent experiments (n = 3). aP < 0.05 versus control, bP < 0.05 versus model].

Table 1 Comparison of the levels of FBG, C-P, TNF-α, IL-6 in different groups ($\bar{x}\pm s$)

Group	n	FBG (mmol/L)	C-P (ng/mL)	TNF-α (ng/mL)	IL-6 (ng/mL)
N	8	5.46±0.49^a	1.79±0.22^a	149±25.92^a	49±10.45^a
M	8	20.48±1.48^b	0.84±0.25^b	238±37.18^b	135±16.53^b
A1	8	17.58±2.74^bc	1.15±0.38^b	194±33.45^c	96±18.42^ab
A2	8	15.65±1.51^ab	1.34±0.51^c	176±38.16^a	79±19.13^ab
A3	8	13.89±3.18^ab	1.59±0.36^a	168±29.13^a	71±21.76^ad
x	8	13.94±1.58^ab	1.67±0.31^a	185±34.43^a	80±19.24^ab

Table 2 Comparison of the levels of blood glucose during OGTT in different groups ($\bar{x}\pm s$)

Group	n	Blood glucose
Group	n	0 h	0.5 h	1 h	2 h	AUC
N	8	5.54±1.89^a	7.55±1.47^a	7.17±1.40^a	5.70±1.39^a	12.92±1.69^a
M	8	20.59±1.95^b	31.58±2.14^b	27.72±1.81^b	23.44±2.41^b	51.41±3.51^b
A1	8	17.63±1.69^ab	28.35±2.97^bc	24.37±0.88^ab	19.22±1.57^ab	44.18±1.94^ab
A2	8	15.31±2.29^ab	26.59±3.14^ab	24.16±2.34^ab	17.39±2.19^ab	41.63±3.05^ab
A3	8	13.47±2.46^ab	23.40±2.93^ab	18.42±2.31^ab	11.47±1.57^ab	31.63±2.97^ab
x	8	13.56±0.95^ab	24.39±1.48^ab	19.30±1.83^ab	12.26±2.40^ab	33.15±3.75^ab

Figure 3 AS-IV ameliorates ROS accumulation in HepG2 cells A: AS-IV reduced insulin-increased intracellular ROS levels in HepG2 cells. B: ROS levels were quantified. n = 3, aP < 0.05 vs the control group, bP < 0.05 vs the insulin resistant group. N: control group, given conventional feed without other interfering factors. M: model group, given high sugar and fat feed but no other interfering factors; ROZ: positive group (30 μM rosiglitazone). ROS: reactive oxygen species.

Figure 4 Effects of AS-IV on regulation of JNK-AKT-GSK3β signaling pathway in insulin-resistant HepG2 cells A: the expressions of JNK and phosphorylated JNK (p-JNK) in insulin-resistant HepG2 cells. B: Relative p-JNK levels were quantified. C: the expressions of PDK1 in insulin-resistant HepG2 cells. D: relative PDK1 levels were shown. E: the expressions of AKT in insulin-resistant HepG2 cells. F: relative AKT levels were quantified. G: the expressions of GSK3β in insulin-resistant HepG2 cells. H:relative GSK3β levels were displayed. Data are expressed as means ± SD from three individual experiments. aP < 0.001, dP < 0.05, fP < 0.01, compared to normal group; bP < 0.05, eP < 0.01, cP < 0.001 compared to model group. N: control group; M: model group; ROZ: rosiglitazone group (30 μM).

References 25

1	Riddy DM, Delerive P, Summers RJ, Sexton PM, Langmead CJ. G Protein-Coupled Receptors Targeting Insulin Resistance, Obesity, and Type 2 Diabetes Mellitus. Pharmacol Rev 2018; 70: 39-67. DOI PMID
2	Farrokhian A, Bahmani F, Taghizadeh M, et al. Selenium supplementation affects insulin resistance and serum hs-CRP in patients with type 2 diabetes and coronary heart disease. Horm Metab Res 2016; 48: 263-8. DOI PMID
3	Motiani P, Virtanen KA, Motiani KK, et al. Decreased insulin-stimulated brown adipose tissue glucose uptake after short-term exercise training in healthy middle-aged men. Diabetes Obes Metab 2017; 19: 1379-88. DOI PMID
4	Keane KN, Cruzat VF, Carlessi R, de Bittencourt PIH, Newsholme P. Molecular events linking oxidative stress and inflammation to insulin resistance and β-cell dysfunction. Oxid Med Cell Longev 2015; 181643.
5	Bonnard C, Durand A, Peyrol S, et al. Mitochondrial dysfunction results from oxidative stress in the skeletal muscle of diet-induced insulin-resistant mice. J Clin Invest 2008; 118: 789-800. DOI PMID
6	Rehman K, Akash MSH. Mechanism of generation of oxidative stress and pathophysiology of type 2 diabetes mellitus: how are they interlinked? J Cell Biochem 2017; 118: 3577-85. DOI PMID
7	Badeau RM, Honka M-J, Lautamäki R, et al. Systemic metabolic markers and myocardial glucose uptake in type 2 diabetic and coronary artery disease patients treated for 16 weeks with rosiglitazone, a PPARγ agonist. Ann Med 2014; 46: 18-23.
8	Zhang J, Liu X, Xie XB, Cheng XC, Wang RL. Multitargeted bioactive ligands for PPARs discovered in the last decade. Chem Biol Drug Des 2016; 88: 635-63. DOI PMID
9	Meijun L, Zhicheng L, Bin X, Wei Z, Jianwei C. Review of systematic reviews and Meta-analyses investigating Traditional Chinese Medicine treatment for type 2 diabetes mellitus. J Tradit Chin Med 2016; 36: 555-63. PMID
10	Li L, Hou X, Xu R, Liu C, Tu M. Research review on the pharmacological effects of astragaloside IV. Fund Clin Pharmacol 2017; 31: 17-36. DOI PMID
11	Lv L, Wu SY, Wang GF, et al. Effect of astragaloside IV on hepatic glucose-regulating enzymes in diabetic mice induced by a high-fat diet and streptozotocin. Phytother Res 2010; 24: 219-24. DOI PMID
12	Yin Y, Qi F, Song Z, Zhang B, Teng J. Ferulic acid combined with astragaloside IV protects against vascular endothelial dysfunction in diabetic rats. BioSci Trends 2014; 8: 217-26. PMID
13	Ji TJ, Wang ZY, Zhu YF, et al. Effect of astragaloside Ⅳ in regulating PI3K/Akt/FoxO1 pathway and inhibiting hepatic gluconeogenesis in diabetic rats. Zhong Guo Shi Yan Fang Ji Xue Za Zhi 2020; 26: 78-86.
14	Wang LN, Fang XM, Hu JP, Jia XF, Zhu GQ, Li YJ. Effect of Qidi Xiaoke prescription on C-peptide, tumor necrosis factor-α, interleukin-6, and histopathological changes of the spleen in rats with type 2 diabetes with deficiency of both Qi and Yin. J Anhui Univ Chin Med 2017; 36: 72-5.
15	Li M, Yang CY, Fu CH, Xiong YG, Ling Y. Study of purslane/ licorice/dandelion compound on insulin resistance in type 2 diabetic rats. Acta Lab Anim Sci Sin 2020: 1-10.
16	Liu P, Shi L, Cang X, et al. CtBP2 ameliorates palmitate-induced insulin resistance in HepG2 cells through ROS mediated JNK pathway. Gen and Comp Endocr 2017; 247: 66-73.
17	Lee BC, Lee J. Cellular and molecular players in adipose tissue inflammation in the development of obesity-induced insulin resistance. BBA - Mol Basis Dis 2014; 1842: 446-62.
18	Dai B, Wu Q, Zeng C, et al. The effect of Liuwei Dihuang decoction on PI3K/Akt signaling pathway in liver of type 2 diabetes mellitus (T2DM) rats with insulin resistance. J Ethnopharmacol 2016; 192: 382-9. DOI PMID
19	Zhang H, Ge Z, Tang S, Meng R, Bi Y, Zhu D. Erythropoietin ameliorates PA-induced insulin resistance through the IRS/AKT/ FOXO1 and GSK-3β signaling pathway, and inhibits the inflammatory response in HepG2 cells. Mol Med Rep 2017; 16: 2295-301.
20	Gao W, Du X, Lei L, et al. NEFA-induced ROS impaired insulin signalling through the JNK and p38MAPK pathways in non-alcoholic steatohepatitis. J Cell Mol Med 2018; 22: 3408. DOI PMID
21	Dong Y, Gao G, Fan H, Li S, Li X, Liu W. Activation of the liver X receptor by agonist TO901317 improves hepatic insulin resistance via suppressing reactive oxygen species and JNK Pathway. PLoS One 2015; 10: e0124778.
22	Yang Y, Li W, Li Y, Wang Q, Gao L, Zhao J. Dietary Lycium barbarum polysaccharide induces Nrf2/ARE pathway and ameliorates insulin resistance induced by high-fat via activation of PI3K/AKT sSignaling. Oxid Med Cell Longev 2014; 145641.
23	He S, Peng WB, Zhou HL. Combination treatment of deep sea water and fucoidan attenuates high glucose-induced insulin-resistance in HepG2 hepatocytes. Mar Drugs 2018; 16: 48.
24	Cordero-Herrera I, Martín MÁ, Goya L, Ramos S. Cocoa flavonoids attenuate high glucose-induced insulin signalling blockade and modulate glucose uptake and production in human HepG2 cells. Food Chem Toxicol 2014; 64: 10-9.
25	Song R, Wang X, Mao Y, et al. Resistin disrupts glycogen synthesis under high insulin and high glucose levels by down-regulating the hepatic levels of GSK3β. Gene 2013; 529: 50-6.

[1]	CHANG Fengjin, ZHOU Peng, LI Guoying, ZHANG Weizhi, ZHANG Yanyan, PENG Daiyin, CHEN Guangliang. Taohong Siwu decoction (桃红四物汤) ameliorates atherosclerosis in rats possibly through toll-like receptor 4/myeloid differentiation primary response protein 88/nuclear factor-κB signal pathway [J]. Journal of Traditional Chinese Medicine, 2024, 44(1): 103-112.
[2]	SUN Qianhui, CHENG Kai, DAI Xingye, YANG Zhiwen, WU Xiaoling, XU Chang, QIU Xinghua, GAO Xiaofeng, LIU Daonan, YANG Qirui. Effect of electroacupuncture at Neiguan (PC6) at different time points on myocardial ischemia reperfusion arrhythmia in rats [J]. Journal of Traditional Chinese Medicine, 2024, 44(1): 113-121.
[3]	LIU Huihui, FENG Jun, LIU Jianhe, CHENG Choufu, HU Guoheng. Efficacy of Jiangzhi Xiaoban tablet (降脂消斑片) on toll-like receptor 4/nuclear factor-kappa B/nod-like receptor protein 3 signaling pathway in mice with atherosclerosis induced by high-fat diet [J]. Journal of Traditional Chinese Medicine, 2024, 44(1): 88-94.
[4]	LIU Yue, ZHANG Fan, HAN Xiaomeng, XU Ningyang, ZHAO Yu, WANG Qige, WANG Jianan, LU Bingjiu, Zhang Yan. Jianpi Qutan Fang (健脾祛痰方) induces anti-atherosclerosis and ameliorates endothelial cell injury in high-fat diet rats via an anti-inflammatory and inhibiting Janus kinase/signal transducer and activator of transcription signaling pathway [J]. Journal of Traditional Chinese Medicine, 2023, 43(6): 1168-1175.
[5]	QIN Xihui, PANG Jianli, XIONG Guan, FENG Jie. Bo′s abdominal acupuncture improves disordered metabolism in obese type 2 diabetic rats through regulating fibroblast growth factor 21 and its related adipokines [J]. Journal of Traditional Chinese Medicine, 2023, 43(6): 1200-1208.
[6]	LI Zhihao, HAN Wenjun, SONG Xiuling, LI Yan, CHEN Yuelai. Electroacupuncture stimulating Zhongji (CV3), Guanyuan (CV4), and bilateral Dahe (KI12) attenuates inflammation in rats with chronic nonbacterial prostatitis induced by estradiol through inhibiting toll-like receptor 4 pathway [J]. Journal of Traditional Chinese Medicine, 2023, 43(5): 963-972.
[7]	HAN Yunpeng, YU Wentao, ZHANG Ying, XU Huazhou, DENG Guoxing, FANG Chaoyi. Qinghua decoction (清化饮) improves chronic nonbacterial prostatitis possibly via regulating the chromogranin A/nerve growth factor/tyrosine kinase A signaling pathway mediated by inflammatory factors [J]. Journal of Traditional Chinese Medicine, 2023, 43(4): 695-703.
[8]	WAN Feng, YANG Ruchun, TANG Yuewen. Uncovering pharmacological mechanisms of Phellinus linteus on focal segmental glomeruloscleosis rats through tandem mass tag-based quantitative proteomic analysis, network pharmacology analysis and experimental validation [J]. Journal of Traditional Chinese Medicine, 2023, 43(4): 744-750.
[9]	PANG Fengtao, LI Kesong, ZHANG Yi, TANG Xiaopo, ZHOU Xinyao. Efficacy of Lushi Runzao decoction (路氏润燥汤) on ameliorating Sjogren's syndrome: a network pharmacology and experimental verification-based study [J]. Journal of Traditional Chinese Medicine, 2023, 43(4): 751-759.
[10]	Minh Duc Nguyen, Thanh Van Tran, Quoc Vinh Nguyen, Cay Doan Ha, Linh Vu Phuong Dang. Effectiveness of bee venom acupuncture for patients suffering from periarthritis humeroscapularis [J]. Journal of Traditional Chinese Medicine, 2023, 43(4): 795-800.
[11]	SHENG Song, ZHANG Yanhong, GAO Hongyang, MA Hangkun, HUANG Ye, LI Qingna, GAO Rui, XU Fengqin. Effectiveness of acupoint application of Xiaozhong Zhitong Tie on diarrhea in patients: a retrospective cohort study in China [J]. Journal of Traditional Chinese Medicine, 2023, 43(4): 809-814.
[12]	LUO Xin, XIE Jing, HUANG Li, GAN Wenfan, CHEN Ming. Efficacy and safety of activating blood circulation and removing blood stasis of Traditional Chinese Medicine for managing renal fibrosis in patients with chronic kidney disease: a systematic review and Meta-analysis [J]. Journal of Traditional Chinese Medicine, 2023, 43(3): 429-440.
[13]	ZHAO Ye, WANG Xian, GU Ling, LI Zihang, ZHU Jingtian, WANG Wenkai, ZHANG Liang, XUE Mei. Efficacy of Danggui Buxue decoction (当归补血汤) on diabetic nephropathy-induced renal fibrosis in rats and possible mechanism [J]. Journal of Traditional Chinese Medicine, 2023, 43(3): 507-513.
[14]	YE Yujuan, WEI Yuting, JIA Jing, YAN Xingke. Efficacy of needling Baihui (GV20), Neiguan (PC6), Shenmen (HT7) and Taichong (LR3) on cerebral cortical blood oxygen level in rats with insomnia [J]. Journal of Traditional Chinese Medicine, 2023, 43(3): 523-532.
[15]	JIA Lihua, KUANG Haodan, XU Yuan. Efficacy of Buzhong Yiqi decoction (补中益气汤) on benign prostatic hyperplasia and its possible mechanism [J]. Journal of Traditional Chinese Medicine, 2023, 43(3): 533-541.

Home

Online First

Author Center

Reviewer Center

Issues

Announcement

About Us

Astragaloside IV ameliorates insulin induced insulin resistance in HepG2 cells through reactive oxygen species mediated c-Jun N-terminal kinase pathway

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 6

References 25

Related Articles 15

Recommended Articles

Metrics

Comments