ABSTRACT
Objective To investigate the mechanism of homocysteine-induced microglia (BV-2 cells) expression of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α).Methods The BV-2 cells were divided into blank control group,cysteine (Cys) group,homocysteine (Hcy) group and homocysteine and glutathione (Hcy+GSH) group,and the BV-2 cells in these groups were incubated with cysteine or homocysteine or homocysteine and glutathione together for 72 h.The mRNA expressions of IL-1β and TNF-α were assessed by RT-qPCR.The protein expressions of IL-1β and TNF-α in supernatant were detected by enzyme linked immunosorbent assay (ELISA).Western blot was used to observe the changes of NF-κB/p65 expression.Results There were significant differences in mRNA and protein expressions of IL-1β and TNF-α and NF-κB/p65 protein expression between groups (F=48.63,130.76,702.91,293.69,212.06,respectively,all P=0.000).The secretions of IL-1β and TNF-α were significantly improved by homocysteine (P＜0.05),and were reversed by the treatment with glutathione (P＜0.05).Western blot assay result showed that NF-κB/p65 was elevated after treatment with homocysteine (P＜0.05).Conclusions Homocysteine can induce microglia expression of interleukin-1β and tumor necrosis factor-α,and NF-κB signaling pathway may be involved in this process.
ABSTRACT
<p><b>OBJECTIVE</b>Inflammatory factors have been known to induce nerve cells apoptosis and decrease learning capacity of diabetics. The aim of this study is to evaluate the inhibitory effect of Gastrodine on the expression of interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6) in culturing for gitter cells (BV-2 cells) induced by high concentration of glucose.</p><p><b>METHOD</b>The BV-2 cells incubated in vitro with different concentrations of glucose and gastrodine were divided into five groups: control group (glucose: 25 mmol x L(-1)), high concetration of glucose (glucose: 45 mmol x L(-1) HCG) group and Gastrodine groups (glucose 45 mmol x L(-1) with gastrodine 25 mg x L(-1) (LG), 50 mg x L(-1) (MG), 100 mg x L(-1) (HG). After culturing for 24 h, morphological changes of cells were observed by inverted phase contrast microscope. The supernatant protein of IL-1 beta and IL-6 was detected by ELISA. The mRNA expression of IL-1 beta and IL-6 was assessed by Reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULT</b>The cells were proned to aggregate, some of them with hypertrophy, distinct nucleoli and branch-shaped hyperplasy in HCG group, while less change in Gastrodine groups. The supernatant protein of IL-1 beta is higher in HCG group than control group (119.53 +/- 15.91) ng x L(-1) vs (25.74 +/- 15.72) ng x L(-1) (P < 0.01), but lower in the gastrodine groups than HCG LG (99.32 +/- 19.66) ng x L(-1), MG (76.94 +/- 17.16) ng x L(-1), HG (88.35 +/- 18.72) ng x L(-1) vs (119.53 +/-15.91) ng x L(-1) (P < 0.05). The supernatant protein of IL-6 protein also higher in HCG than control group (393.7 +/- 17.51) ng x L(-1) vs (125.85 +/- 36.62) ng x L(-1) (P < 0.01), and lower in the gastrodine groups than HCG (LG 327.06 +/- 23.53) ng x L(-1), MG (217.36 +/- 28.81) ng x L(-1), HG (263.17 +/- 22.32) ng x L(-1) vs (393.7 +/- 17.51) ng x L(-1), P < 0.05). The mRNA expression of IL-1 beta was increased significantly higher in HCG than control group (2.77 +/- 0.29) vs (1.13 +/- 0.27) (P < 0.05), but decreased significantly in gastrodine groups than HCG LGA (2.66 +/- 0.31), MGA (2.1 +/- 0.41), HGA (2.4 +/- 0.28) vs (2.77 +/- 0.29) (P < 0.05). The mRNA Expression of IL-6 was higher in HCG than control group (3.97 +/- 0.33) vs (1.05 +/- 0.13) (P < 0.05, but lower in gastrodine groups than HCG LG (3.28 +/- 0.3), MG (2.65 +/- 0.33), HG (3.04 +/- 0.26), vs (3.97 +/- 0.33) (P < 0.05).</p><p><b>CONCLUSION</b>Gastrodine can inhibit the expression of IL-1 beta, IL-6 in cultured BV-2 cells induced by high concentration of glucose.</p>