China Animal Husbandry and Veterinary Medicine ›› 2023, Vol. 50 ›› Issue (7): 2820-2831.doi: 10.16431/j.cnki.1671-7236.2023.07.022

• Preventive Veterinary Medicine • Previous Articles     Next Articles

Catalpol Alleviates the Damaging Effect of AGEs on the Endothelial Cells in the Hepatic Sinus by Disrupting the Interaction Between Galectin-3 and CD146

SUN Weixiang1,2,3, ZONG Yingying6, LIU Bo1, ZHUANG Yuan1, QIN Feng1,2, CHEN Yu1, ZHU Shanyuan1,2,3, XU Huiqin4,5   

  1. 1. Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China;
    2. Jiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Taizhou 225300, China;
    3. Engineering Technology Research Center for Modern Animal Science and Novel Veterinary Pharmaceutic Development, Taizhou 225300, China;
    4. School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China;
    5. Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica, Nanjing 210023, China;
    6. Nanjing University of Chinese Medicine Hanlin College, Taizhou 225300, China
  • Received:2022-10-31 Published:2023-06-30

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Abstract: 【Objective】 The purpose of this study was to investigate the effect of catalpol on the inflammatory injury of rat liver sinusoidal endothelial cells (RLSECs) induced by advanced glycation end products (AGEs) via affecting the interaction between Galectin-3 and CD146.【Method】 After incubating RLSECs with 0, 0.1, 1 and 10 μmol/L catalpol for 48 h, the effect of cell proliferation was observed by CCK-8 method.Incubating RLSECs with 10 μmol/L catalpol for 0, 12, 24, 48 and 96 h, and the effect of cell proliferation was observed by the same method as above.Set Control (blank control group), AGEs (AGEs treatment), Cat1 (1 μ mol/L catalpol), Cat10 (10 μ mol/L catalpol) and positive control GB1107 (1 μmol/L GB1107) groups.After Cat1, Cat10 and GB1107 groups were incubated with drugs for 30 min, then all groups were stimulated by 200 μg/mL AGEs except for Control group, the morphological changes of RLSECs in the above groups were observed, and the degree of cell damage was detected by lactate dehydrogenase (LDH) method.The secretion of monocyte chemoattractant protein-1 (MCP-1) and intercellular adhesion molecule-1 (ICAM-1) were measured by ELISA.The macrophages RAW264.7 were divided into groups and administered with the same method as above.After 48 h, the amount of nitric oxide (NO) released to the cell supernatant in each group was observed with Griess method.RAW264.7 cells were seeded in the Transwell chamber and RLSECs were seeded at the bottom of the well plate.Control, AGEs, Cat10, Cat10+LV Galectin-3-GFP and Cat10+LV Galectin-3-shRNA groups were set up.The last two groups were transfected with lentivirus for 48 h, and then administered with drugs for 30 min respectively, AGEs were added to the culture medium with a final concentration of 200 μg/mL for stimulation except for Control group, and the number of migrated macrophages was observed by crystal violet method 48 h later.After incubation of RLSECs in Control, AGEs, Cat10 and GB1107 groups for 48 h, the co-localization of Galectin-3 and CD146 was observed by immunofluorescence.The protein samples from Control, AGEs and Cat10 groups were used to detect the interaction between Galectin-3 and CD146 and their respective expression levels by Western blotting and Co-IP methods.【Result】 Compared with Control group, there was no significant difference in cell proliferation rate of RLSECs in Cat0.1 group (P>0.05), while the cell proliferation rate of RLSECs in Cat1 and Cat10 groups was significantly increased (P<0.05).Compared with the group incubated for 0 h, the cell profiferation rate of RLSECs was significantly increased in the group with 10 μmol/L catalpol incubated for 48 h. Therefore, cells were incubated with 10 μmol/L catatpol for 48 h for subsequent experiments. Compared with AGEs group, in the Cat1 and Cat10 groups the cell damage induced by AGEs was improved, and the LDH activity of RLSECs supernatant, the release of MCP-1 and ICAM-1 were significantly decreased (P<0.05).Compared with AGEs group, in the catalpol groups the activation of macrophage RAW264.7 was inhibited and the release of NO from cells was significantly reduced (P<0.05).By overexpression of lentivirus vector and knockdown of Galectin-3 of RLSECs and RAW264.7, it was proved that catalpol could significantly improve the infiltration of macrophages by inhibiting the expression of this molecule (P<0.05).Compared with AGEs group, in Cat10 group the binding of Galectin-3 and CD146 was significantly inhibited (P<0.05) and the respective expression of the two molecules did not affect (P>0.05).【Conclusion】 Catalpol could promote the uncoupling effect of Galectin-3 and CD146 molecular complexes induced by AGEs, improved the damage of sinusoidal vascular endothelial cells induced by AGEs and the release of proinflammatory factors, reduced the activation of macrophages and the secretion of NO, and played a protective role in sinusoidal vascular endothelial cells.The mechanism of catalpol to improve the liver injury caused by AGEs deposition in diabetes was preliminarily proved through in vitro experiments.

Key words: catalpol; diabetic liver injury; advanced glycation end products(AGEs); Galectin-3; CD146; inflammatory injury

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