Effects of curcumin on pro-inflammatory factors in pulmonary microvascular endothelial cell thrombus model stimulated by thrombus / 中国中西医结合急救杂志

ABSTRACT

Objective To explore the effects of curcumin on pro-inflammatory factors in the lung microvascular endothelial cells (LMVEC) model stimulated by thrombus. Methods The LMVECs were divided into six groups according to the random number table method. No treatment was given to the blank control group ; the model group was cultured for 7 hours in normal medium; the curcumin group was treated with 40 μmol/L curcumin for 72 hours ; the shRNA group was infected with shRNA adenovirus for 72 hours; the irregular chemokines (CX3CL1) overexpression group was infected with CX3CL1 overexpressing adenovirus for 72 hours; the shRNA+curcumin group infected with shRNA adenovirus and treated with 40 μmol/L curcumin together for 72 hours; CX3CL1 overexpression +curcumin group infected with CX3CL1 overexpressing adenovirus and treated with 40 μmol/L curcumin together for 72 hours. After each group was given the corresponding pretreatment, the thrombus natural precipitation was added each group for 12 hours. The contents of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), the mRNA expression levels of CX3CL1, CX3CL1 receptor (CX3CR1), IL-6, TNF-α and the protein expression levels of CX3CL1/CX3CR1, CX3CR1/NF-κB in various groups were observed, repeat 3 times in each group. Results The contents and mRNA expression of IL-6, TNF-αand protein expression of CX3CR1, NF-κB in the LMVEC group were significantly higher than those in blank control group [IL-6 (ng/L): 207.90±16.69 vs. 85.93±20.32, TNF-α (ng/L): 239.60±15.27 vs. 101.23±11.92; IL-6 mRNA: 0.66±0.05 vs. 0.11±0.02, TNF-α mRNA: 1.06±0.04 vs. 0.02±0.01; CX3CR1 protein:3.94±0.58 vs. 1.00±0.31, NF-κB protein: 1.20±0.07 vs. 1.00±0.10; all P < 0.05]; the contents of IL-6 in shRNA group, CX3CL1 overexpression group, shRNA + curcumin group, CX3CL1 overexpression + curcumin group were all obviously lower than those in LMVEC group (ng/L: 183.60±11.52, 159.27±15.02, 117.03±7.91, 119.97±11.43 vs. 207.90±16.69, all P < 0.01); the content of TNF-α was markedly increased in shRNA group compared with that of LMVEC group (ng/L: 282.00±5.63 vs. 239.6±15.27), while the contents of TNF-α in CX3CL1 overexpression group, shRNA+ curcumin group, CX3CL1 overexpression + curcumin group were all lower than those in LMVEC group (ng/L: 216.97±9.20, 203.97±19.03, 191.97±17.50 vs. 239.6±15.27, all P < 0.05). The mRNA expression levels in CX3CL1 overexpression group and CX3CL1 overexpression + curcumin group were significantly higher than those in the blank control group and the LMVEC group (CX3CL1 mRNA: 55 210.3±1 209.2, 165 296.3±8 082.4 vs. 3.3±0.6, 2.0±0.0, all P < 0.01). The mRNA expression level of IL-6 in shRNA group was higher than that in LMVEC group (0.82±0.17 vs. 0.66±0.05), the mRNA expression level of IL-6 in CX3CL1 overexpression was lower than that in LMVEC group (0.29±0.03 vs. 0.66±0.05), the changes after pretreatment with curcumin were more significant (1.06±0.03 vs. 0.66±0.05 and 0.15±0.01 vs. 0.66±0.05); the mRNA expressions of TNF-α in shRNA group, CX3CL1 overexpression group, shRNA+ curcumin group were significantly lower than those in LMVEC group (0.41±0.04, 0.88±0.07, 1.01±0.02 vs. 1.06±0.04), the mRNA expression level of TNF-α in CX3CL1 overexpression + curcumin group was significantly higher than that in LMVEC group (1.36±0.01 vs. 1.06±0.04). The protein expression of CX3CL1, CX3CR1, NF-κB in shRNA group, CX3CL1 overexpression group, shRNA + curcumin group, CX3CL1 overexpressing + curcumin group were significantly higher than those in the LMVEC group (CX3CL1 protein: 0.41±0.07, 0.59±0.09, 0.69±0.61, 1.02±0.23 vs. 1.33±0.33, CX3CR1 protein: 0.85±0.18, 1.10±0.16, 1.32±0.18, 1.54±0.08 vs. 3.94±0.58, NF-κB protein: 0.33±0.07, 0.41±0.08, 0.41±0.07, 0.63±0.08 vs. 1.20±0.07). Conclusion Curcumin can inhibit the secretion of IL-6, TNF-α, CX3CR1 and NF-κB in thrombus-stimulated LMVEC model.