Abstract
Yunvjian (YNJ) is a traditional Chinese medicine formula adopted to prevent and treat diabetes. Our previous results from animal experiments showed that YNJ decreased blood glucose. This study aimed to examine the effect of high glucose and high lipid (HG/HL) conditions on the proliferation and apoptosis of INS-1 cells and the possible protective mechanism of YNJ-medicated serum on INS-1 cells exposed to HG/HL conditions. INS-1 cells were cultured in RPMI 1640 medium after being passaged. Then, INS-1 cells in the logarithmic growth phase were collected and divided into five groups: control, HG/HL, HG/HL+5% YNJ-medicated serum, HG/HL+10% YNJ-medicated serum, and HG/HL+20% YNJ-medicated serum. MTT assay and flow cytometry were used to detect proliferation and apoptosis of INS-1 cells, respectively. Protein profiles of INS-1 cells were analyzed using a tandem mass tag (TMT) label-based quantitative proteomic approach. Western blotting was performed to verify the proteomic results. YNJ-medicated serum significantly promoted INS-1 cell proliferation and inhibited apoptosis. Proteomic results from the INS-1 cells in the control, HG/HL, and HG/HL+10% YNJ-medicated serum groups showed that 7,468 proteins were identified, of which 6,423 proteins were quantified. Compared with the HG/HL group,430 differential proteins were upregulated, and 671 were downregulated in the HG/HL+10% YNJ-medicated serum group. Compared with the control group, 711 differential proteins were upregulated and 455 were downregulated in the HG/HL group, whereas 10 differential proteins were upregulated and 9 were downregulated in the HG/HL+10% YNJ-medicated serum group. Furthermore, several proteins related to autophagy, including ATG3, ATG2B, GABARAP, WIPI2, and p62/SQSTM1, were verified by western blotting, and these results were consistent with the results obtained from the proteomics analysis. These results confirmed that the autophagy pathway is critical to glucolipotoxicity in INS-1 cells. YNJ-medicated serum exhibited a protective effect on INS-1 cells cultured under HG/HL conditions by regulating autophagy genes' expression and restoring the autophagic flux.
Highlights
Autophagy can clear misfolded proteins and damaged organelles produced in cell growth, metabolism, and stress, and timely removal of these substrates is essential for cells to maintain homeostasis and normal physiological activities
Different concentrations of YNJ-medicated serum exhibit different effects on the proliferation of INS-1 cultured under high glucose (HG)/HL condition for 48 h. e at 590 nm (A590) values in the control, HG and high lipids (HG/HL), high glucose and high lipid (HG/ HL)+5%, HG/HL+10%, and HG/HL+20% YNJ-medicated serum groups were 2.38 ± 0.17, 1.75 ± 0.32, 2.11 ± 0.09, 2.35 ± 0.16, and 2.09 ± 0.13, respectively. ere were statistically significant differences in the A590 values between the control and HG/HL groups (p < 0.01), HG/HL and HG/HL+5%YNJ-medicated serum groups (p < 0.05), HG/HL and HG/HL+10 %YNJ-medicated serum groups (p < 0.01), and HG/HL and HG/HL+20 %YNJ-medicated serum groups (p < 0.05). e results suggested that HG/ HL could reduce cell proliferation after 48 h of culture
All 5%, 10%, and 20% YNJ-medicated serum promoted the proliferation of INS-1 cells cultured under HG/HL conditions, and 10% concentration of YNJ-medicated serum had a better effect than 5% and 20% concentration of YNJmedicated serum (Table 2)
Summary
Autophagy can clear misfolded proteins and damaged organelles produced in cell growth, metabolism, and stress, and timely removal of these substrates is essential for cells to maintain homeostasis and normal physiological activities. Studies have found that autophagy is the cell survival mechanism; it participates in the occurrence and development of various diseases, such as tumors, diabetes, liver injury, and leukemia. Β-cell dysfunction and apoptosis are Evidence-Based Complementary and Alternative Medicine the two major characteristics of the late stages of Type 2 diabetes mellitus (T2D) [2]. With prolonged exposure to glucolipotoxicity, the body’s selfcompensation function gradually decreases or disappears; irreversible damage to pancreatic β-cell function occurs, including reduced β-cell sensitivity to glucose, insufficient insulin secretion, and β-cell apoptosis [4], thereby inducing the occurrence of T2D. Us, glucolipotoxicity mediated β-cell loss is a critical causal factor in the late stages of diabetes; the exact mechanisms remain unclear With prolonged exposure to glucolipotoxicity, the body’s selfcompensation function gradually decreases or disappears; irreversible damage to pancreatic β-cell function occurs, including reduced β-cell sensitivity to glucose, insufficient insulin secretion, and β-cell apoptosis [4], thereby inducing the occurrence of T2D. us, glucolipotoxicity mediated β-cell loss is a critical causal factor in the late stages of diabetes; the exact mechanisms remain unclear
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