补阳还五汤对脑缺血大鼠脑组织核转录因子-κBp65及其抑制物表达的影响

摘要

Objective To explore the effects of Buyang Huanwu decoction(BYHWD)on expressions of nuclear factor-κB p65(NF-κBp65)and its inhibitor( I-κB)in signal transduction of NF-κB in brain tissue of rats with focal cerebral ischemia injury. Methods 180 Sprague-Dawley(SD)rats were randomly divided into normal group,sham-operated group,model group,pynolidine dithiocarbamate(PDTC)group,minocycline(MC)group and BYHWD treatment group,each group 30 rats. The rats of PDTC group were given PDTC 100 mg�kg-1�d-1 by intra-peritoneal injection. In MC group,MC was given by filling the stomach,the dose was 2.35 g�kg-1�d-1,the drug solution was prepared by adding the distilled water,and the total volume of drug solution to fill the stomach was kept at the same volume in various groups,thus the concentration of the drug was different. In BYHWD group,BYHWD was given,the dose was reduced to 5 g�kg-1�d-1 according to the body surface area dose conversion formula about people and animals. In sham-operated group and model group,the distilled water was given in the same volume as other drug solution. The protein expression levels of NF-κBp65 and I-κB in ischemic tissues were examined by using immunohistochemical method on the time points 7,14 and 21 days after treatment in each group. Results Compared with model group, the cell numbers with expression of NF-κBp65 in PDTC group,MC group and BYHWD group were significantly decreased along with the prolongation of therapy time,the decrease in number was more and more,until 21 days,it reached the valley level(cell/400 times HP:44.00±6.91,45.33±6.55,18.67±2.14 vs. 126.00±5.78,all P<0.05);the number of cells with expression of I-κB was obviously increased,the differences being statistically significant(all P<0.05),but the differences in expression of NF-κBp65 among the treatment groups at the different time points were not statistically significant(all P>0.05). After treatment for 7 days,the number of cells with positive expression of I-κB protein in BYHWD group was less than that in MC group(cell/400 times HP:55.00±3.40 vs. 72.50±4.29,P<0.05);after treatment for 14 days,the number in BYHWD group was approximately the same as that in the MC group, the difference being not statistically significant(93.50±6.15 vs. 93.00±6.20,P>0.05),and after treatment for 21 days,the number in BYHWD group was significantly higher than that in MC group(88.83±4.95 vs. 71.17±7.16, P<0.05). Conclusion BYHWD can regulate the expressions of inflammatory cytokine I-κB and NF-κB in signal transduction of NF-κB in ischemic brain tissue to inhibit the inflammatory reaction,thus it has the protective effect on cerebral ischemia.%目的：探讨补阳还五汤对脑缺血大鼠脑组织核转录因子-κB（NF-κB）信号通路NF-κBp65及其抑制物（I-κB）表达的影响。方法将180只SD大鼠按随机数字表法分为正常对照组、假手术组、模型组、吡咯烷二硫代氨基甲酸盐（PDTC）组、米诺环素和补阳还五汤组，每组30只。PDTC组腹腔注射100 mg�kg-1�d-1 PDTC；米诺环素组给予米诺环素2.35 g�kg-1�d-1加蒸馏水按相同体积配成各浓度药液灌胃；补阳还五汤组给予补阳还五汤，按人与动物体表面积折算为5 g�kg-1�d-1；假手术组和模型组给予等体积蒸馏水。采用免疫组化法检测各组给药后7、14、21 d缺血脑组织NF-κBp65和I-κB蛋白的表达情况。结果与模型组比较，PDTC组、米诺环素组和补阳还五汤组NF-κBp65阳性表达细胞数随时间延长依次减少，21 d达谷值（个/400倍视野：44.00±6.91、45.33±6.55、18.67±2.14比126.00±5.78，均P＜0.05）；I-κB阳性表达细胞数显著增加，差异均有统计学意义（均P＜0.05）；而各用药组不同时间点NF-κBp65比较差异均无统计意义（均P＞0.05）。给药后7 d，补阳还五汤组I-κB阳性表达细胞数明显低于米诺环素组（个/400倍视野：55.00±3.40比72.50±4.29， P＜0.05）；14 d时接近米诺环素组（93.50±6.15比93.00±6.20，P＞0.05）；21 d时已明显高于米诺环素组（88.83±4.95比71.17±7.16，P＜0.05）。结论补阳还五汤可通过调控缺血脑组织NF-κB信号通路炎症因子NF-κBp65和I-κB表达，从而抑制炎症反应来达到对脑缺血的保护作用。