| Objective:To observe influences of D-limonene on brain tissue nuclear factor-?B(NF-?B) and nuclear factor ?B profilin(I?B) expression in rats with alcoholic brain damage, tumor necrosis factor(TNF-?), and interleukin-10(IL-10) level.Methods: Selected 60 male Wistar rats of grade SPF with the weight of 180-200 g, after feeding 7 days adaptively, divided them into 6 groups(n=10) in line with the random number table. The A group: black control group, after giving normal saline to do gavage every day, the top two weeks were 8ml/(kg? d), and later four weeks were 12ml(kg? d). The B group: alcohol model group, After giving 50%(V/V) ethyl alcohol to do gavage every day, the top two weeks were 8ml/(kg? d), and the later four weeks were 12ml(kg? d). The C, D and E three groups were given D-limonene with different concentrations, and dosage was 100, 200 and 400mg/(kg d), respectively, and also given 50%(V/V) alcohol, and dosage was the same to the model group. The F group was the single D-limonene high dose group and was given the dosage of D-limonene of 400mg/(kg? d). After dosing finally, rats were absolute diet but not absolute water for 12 h. They were narcotized with 10% of chloral hydrate and remained brain tissue. The left brain tissue hippocampus used HE staining method t observe its pathological alternation. Immunohistochemical method was used to detect expression situations of NF-?B and I?B in brain tissue and calculate positive cell population. Right brain tissue hippocampus uses ELISA to detect concentrations of TNF-? and IL-10 in brain homogenate.Results:(1) Histopathological observation: After HE dyeing, it can be observed that hippocampus nerve cells in the alcohol group were reduced and had uneven distribution. Nucleus was obviously compacted and transformed and was difficult to distinguish. Hippocampus nerve cells in the black control group and single D-limonene high dose group had clearer arrangement layers, cell structure was complete(round and larger nucleus, clear nuclear membranes and even HE dyeing), individual nerve cells appeared heteromorphism and reflected in karyopyknosis, normally lessened soma, less regular shape, and crimson cytoplasm dyeing. Hippocampus in the limonene with various doses had heteromorphism nerve cells with unequal quantity. Compared with the alcohol, heteroideus nerve cells were reduced obviously.(2) Positive cells of NF-?B and I?B proteins in brain tissue in rats: By making a comparison between the alcohol model group and the black control group, positive cells of NF-?B were increased, while positive cells of I?B were reduced(P<0.05). After giving limonene with different doses, positive cells of NF-?B in brain tissue in rats were gradually reduced with the increase of limonene dose, but positive cells of I?B were gradually increased. Compared with the model group(P<0.05), the black control group and single D-limonene medicine group had no statistical significance.(3) TNF-? expression in brain tissue homogenate in rats: TNF-? expression in brain homogenate in the model group was obviously higher than the control group(P<0.05). TNF-? expression in brain homogenate in rats in the group with lower, middle and high doses of limonene was reduced by comparing with the model group(P<0.05); The single D-limonene medicine group and the black control group had no statistical significance.(4) IL-10 expression in brain tissue homogenate in rats: IL-10 expression in the alcohol model group was higher the black control group(P<0.05); IL-10 expression of brain tissue homogenate in rats in the group with middle and high doses of limonene was obviously higher than the alcohol model group(P<0.05). The black control group and single D-limonene medicine group had no statistical significance.Conclusions: D-limonene can relieve brain tissue damage caused by excessive alcohol. Mechanism of action may be related to inhibiting NF-?B expression, up-regulating I?B expression and the level of regulatory to TNF-? and IL-10. |
PDF Full Text Request