胡黄连苷Ⅱ对大鼠脑缺血再灌注损伤后TLR4、NF-κB及I-κB表达的影响

Effects of Picroside Ⅱ on the Expression of TLR4,NFκB and I-κB in Rats with Cerebral Ischemia-reperfusion Injury

目的观察胡黄连苷Ⅱ对大鼠脑缺血再灌注损伤后Toll样受体4(TLR4)、核因子-κB(NF-κB)及核因子-κB抑制剂(I-κB)表达的影响。方法选取成年雌性健康大鼠72只,从中随机选择12只为空白对照组,其余60只大鼠采用线栓法建立大脑中动脉脑缺血再灌注损伤的模型。选取建模成功的36只大鼠,采用随机数字表法分为胡黄连苷Ⅱ治疗组、阳性对照组以及阴性对照组,每组12只。胡黄连苷Ⅱ治疗组大鼠给予胡黄连苷Ⅱ注射干预,阳性对照组大鼠给予丹参素钠干预,空白对照组和阴性对照组给予磷酸盐缓冲液(PBS)干预。采用TUNEL染色方法检测3组大鼠的细胞凋亡情况;采用免疫组织化学染色方法检测各组大鼠脑内TLR4、NF-κB及I-κB表达的情况。结果空白对照组大鼠的TUNEL阳性细胞呈散在分布、数量较少,大鼠的海马组织、纹状体、大脑皮质中TLR4、NF-κB及I-κB的表达较微弱;阴性对照组大鼠的TUNEL阳性细胞比空白对照组大鼠的数量显著增加,海马组织、纹状体、大脑皮质中TLR4、NF-κB及I-κB的表达较空白对照组显著增加(P<0.05);胡黄连苷Ⅱ治疗组以及阳性对照组大鼠TUNEL阳性细胞数、不同脑组织中TLR4、NF-κB及I-κB的表达均较阴性对照组大鼠低(P<0.05),组间差异无统计学意义(P>0.05)。结论对脑缺血再灌注损伤大鼠来说,给予胡黄连苷Ⅱ治疗可以下调大鼠TLR4、NF-κB及I-κB的表达,显著的抑制大鼠的炎症反应,抑制神经细胞的凋亡。

Objective: To discuss the effects of picroside Ⅱ on the expression of TLR4,NFκB and I-κB in rats with cerebral ischemia-reperfusion injury. Methods: 72 cases of healthy adult female rats were chosen,from which,12 cases were randomly selected as the blank control group,and the rest of the 60 rats were established as middle cerebral artery ischemia-reperfusion injury model with suture method. 36 rats of successful model were selected and randomly divided into 3 groups: the picroside Ⅱ treatment group,the positive control group and the negative control group(12 cases in each) with random number table method. The picroside Ⅱ treatment group was given picroside Ⅱ injection intervention,the positive control group danshensu sodium intervention,the blank control group and negative control group phosphate buffer(PBS) intervention. Apoptosis of the above groups were observed with TUNEL staining method. The expression of TLR4,NFκB and I-κB were observed with immunohistochemical staining method. Results: TUNEL-positive cells of blank control group were scattered,small number,and the expression of TLR4,NFκB and I-κB in hippocampal tissue,striatum,cerebral cortex were relatively weak;TUNEL-positive cells of negative control group significantly increased than the blank control group,and the expression of TLR4,NF-κB and I-κB in hippocampal tissue,striatum,cerebral cortex significantly increased compared with the control group(P < 0.05);TUNEL-positive cells and the expression of TLR4,NF-κB and I-κB of picroside Ⅱ treatment group,positive control group were lower than those in the the negative control group(P <0.05),but there was no statistically significant difference between the two groups(P > 0.05). Conclusion: For the rats with cerebral ischemia reperfusion injury,picroside Ⅱ treatment can cut the expression of TLR4,NF-κB and I-κB,and significantly inhibit the inflammatory response in rats and neuronal apoptosis.