Effects of tripterygium glycosides on high glucose induced apoptosis of human renal tubular epithelial cells and CXCL10/CXCR3 axis

doi:10.11958/20221078

Abstract

Abstract:

Objective To investigate effects of tripterygium glycosides (TG) on the apoptosis of human renal tubular epithelial cells HK-2 induced by high glucose and the CXC chemokine ligand 10 (CXCL10)/CXC chemokine receptor 3 (CXCR3) axis. Methods Human renal tubular epithelial cells HK-2 were cultured in vitro and divided into the control group (culture medium containing 5.5 mmol/L glucose), the high glucose group (culture medium containing 25 mmol/L glucose) and the 12.5, 25 and 50 mg/L TG groups (culture medium containing 12.5, 25 and 50 mg/L TG and 25 mmol/L glucose, respectively). Methyl thiazolyl tetrazolium (MTT) method was used to detect the viability of HK-2 cells. Flow cytometry was used to detect the apoptosis of HK-2 cells. 2',7'-dichlorodi-hydrofluorescein diacetate method was used to detect reactive oxygen species (ROS) level of HK-2 cells. Xanthine oxidase method was used to detect superoxide dismutase (SOD) level of HK-2 cells. Enzyme-linked immunosorbent assay was used to detect levels of inflammatory factors of tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1) in HK-2 cells. Western blot assay was used to detect the expression of apoptotic proteins (Caspase-3, Caspase-9), CXCL10 and CXCR3 proteins in HK-2 cells. Results Compared with the control group, the HK-2 cell viability and SOD level were significantly reduced in the high glucose group, and the apoptosis rate, ROS, TNF-α, TGF-β1 levels and Caspase-3, Caspase-9, CXCL10, CXCR3 protein expression levels were significantly increased (P<0.05). Compared with the high glucose group, the HK-2 cell viability and SOD level were significantly increased in the 12.5, 25 and 50 mg/L TG groups, and the apoptosis rate, ROS, TNF-α, TGF-β1 levels and Caspase-3, Caspase-9, CXCL10, CXCR3 protein expression levels were significantly reduced, and high dose TG has better effect (P<0.05). Conclusion TG can inhibit high glucose induced apoptosis, oxidative stress and inflammation of HK-2 cells, and its mechanism may be related to the inhibition of CXCL10/CXCR3 axis.

Key words: Tripterygium Wilfordii, chemokine CXCL10, receptors, CXCR3, apoptosis, oxidative stress, tripterygium glycosides, high glucose, human renal tubular epithelial cells HK-2

CLC Number:

R587.24

Figures/Tables 7

References 20

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组别	细胞活力	细胞凋亡率
对照组	100.00±0.00	8.34±1.84
高糖组	62.75±3.46^a	36.75±2.73^a
12.5 mg/L TG组	76.42±3.21^b	29.63±2.19^b
25 mg/L TG组	85.14±3.89^bc	21.56±1.95^bc
50 mg/L TG组	93.22±4.16^bcd	11.42±1.54^bcd
F	116.531^＊＊	197.208^＊＊

组别	细胞活力	细胞凋亡率
对照组	100.00±0.00	8.34±1.84
高糖组	62.75±3.46^a	36.75±2.73^a
12.5 mg/L TG组	76.42±3.21^b	29.63±2.19^b
25 mg/L TG组	85.14±3.89^bc	21.56±1.95^bc
50 mg/L TG组	93.22±4.16^bcd	11.42±1.54^bcd
F	116.531^＊＊	197.208^＊＊

组别	ROS（荧光强度）	SOD（U/mL）
对照组	100.49±8.17	16.72±1.96
高糖组	254.39±14.54^a	7.41±0.77^a
12.5 mg/L TG组	207.58±13.47^b	9.12±0.98^b
25 mg/L TG组	162.71±10.32^bc	11.36±1.25^bc
50 mg/L TG组	112.88±9.47^bcd	14.58±1.47^bcd
F	190.452^＊＊	48.180^＊＊

组别	ROS（荧光强度）	SOD（U/mL）
对照组	100.49±8.17	16.72±1.96
高糖组	254.39±14.54^a	7.41±0.77^a
12.5 mg/L TG组	207.58±13.47^b	9.12±0.98^b
25 mg/L TG组	162.71±10.32^bc	11.36±1.25^bc
50 mg/L TG组	112.88±9.47^bcd	14.58±1.47^bcd
F	190.452^＊＊	48.180^＊＊

组别	TNF-α	TGF-β1
对照组	21.44±3.56	25.39±4.37
高糖组	68.79±7.11^a	77.92±7.81^a
12.5 mg/L TG组	55.36±6.68^b	61.43±5.76^b
25 mg/L TG组	35.42±4.45^bc	45.72±5.11^bc
50 mg/L TG组	24.91±3.13^bcd	31.24±4.69^bcd
F	90.086^＊＊	87.014^＊＊