Study on Mechanism of Heat Shock Protein 70 Involved in Intestinal Inflammation through TLR4/MyD88/NF- κB Pathway
WANG Quan-peng，HUANG Chong-jie，CAO Wei-lan
Abstract:
Objective To study the mechanism of heat shock protein 70 involved in intestinal inflammation.Methods Caco-2 cells treated with different lentivirus were divided into HSP70 overexpression group, HSP70 overexpression negative control group, HSP70 low expression group, HSP70 low expression negative control group and blank control group. Lipopolysaccharide stimulated the simulated inflammatory environment. The expression and content of key proteins and cytokines in the signal pathway were determined by Western blotting, reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and ELISA kit, and the cell proliferation rate was measured by EdU colorimetry. Results After 24 hours of LPS stimulation, the mRNA expression and protein content of IL-1 β and TNF- α in Caco-2 cells in each group were significantly higher than those in the group without LPS (P < 0.05). Compared with the low expression group, the mRNA expression of IL-1 β and TNF- α in HSP70 overexpression group were 1.87 and 1.94 times, the protein contents were 1.77 and 1.57 times, and the protein expression of HSP70, Toll-like receptor 4 (TLR4), myeloid differentiation factor 88(MyD88) and nuclear factor-kappa B (NF-κB) were 1.86, 1.96, 2.12, 1.69 times respectively. The relative expression of mRNA were 6.05, 2.98, 3.02, 2.46 times, and the differences were statistically significant.After lipopolysaccharide stimulation, the cell proliferation rate of HSP70 overexpression group was 1.64 times higher than that of overexpression negative control group, and that of HSP70 low expression group was 0.68 times higher than that of overexpression negative control group, and the cell proliferation rate of HSP70 overexpression group was 2.14 times higher than that of low expression group. Conclusion Heat shock protein 70 can produce inflammatory response through TLR4/MYD88/NF-κB inflammatory pathway to participate in the occurrence of intestinal inflammation,which can also promote the proliferation of intestinal cells.
