清热化瘀Ⅱ号方对大鼠脑缺血再灌注损伤TLR3、TRAM mRNA表达的影响  

Influence of Qingre Huayu Formula Ⅱ on expression of TLR3 and TRIFM m RNA in rats with cerebral ischemia reperfusion injury

作　　者：祝美珍[1] 苏春寿 刘泰[1] 肖健[1] 武丽[1] 胡玉英[1] 岑妍慧[1] 贾微[1] 齐玉洁[1] 

机构地区：[1]广西中医药大学,南宁530001

出　　处：《中华中医药杂志》2015年第11期4098-4102,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：广西自然科学基金重点项目(No.2012GXNSFDA276031);国家自然科学基金项目(No.81460725);广西特色实验动物病证模型重点实验室项目(No.J14049);广西高校科学技术研究重点项目(No.ZD2014069)~~

摘　　要：目的:观察清热化瘀Ⅱ号方对大鼠脑缺血再灌注损伤Toll样受体3(TLR3)、TRIF相关受体衔接分子(TRAM)的影响,探讨清热化瘀Ⅱ号方对大鼠脑缺血再灌注损伤神经元保护的作用机制。方法:将192只健康SD大鼠随机分为两大组,每大组再分为4个亚组:空白组、假手术组、脑缺血再灌注组、清热化瘀Ⅱ号方组。采用二次线栓法制备大鼠局灶性脑缺血再灌注模型。假手术组、缺血再灌注组和清热化瘀Ⅱ号方组大鼠分别在术后缺血再灌注2h后3、6、12、24、48h进行标本采集。取材后,常规HE染色并观察各组大鼠脑组织病理形态学改变,应用荧光定量PCR法检测缺血侧海马区TLR3、TRAM m RNA的表达。结果:HE染色:光镜下空白组及假手术组均未见神经元损伤,脑缺血再灌注组及清热化瘀Ⅱ号方组有不同程度的神经元损伤,清热化瘀Ⅱ号方组在各时间点的受损神经元明显少于脑缺血再灌注组。荧光定量PCR结果:空白组与假手术组TLR3、TRAM m RNA呈现低水平表达;脑缺血再灌注组TLR3、TRAM m RNA的表达较假手术组明显增加,差异有统计学意义(P<0.05);脑缺血再灌注组和清热化瘀Ⅱ号方组中,在缺血再灌注3h,TLR3、TRAM m RNA表达开始增加,至24h达高峰,48h后表达逐步下降。清热化瘀Ⅱ号方组TLR3、TRAM m RNA表达活性在同一时间点相比均低于脑缺血再灌注组,差异有统计学意义(P<0.05)。结论:清热化瘀Ⅱ号方可能通过下调TLR3、TRAM m RNA表达,抑制TLRs信号通路的传导,减轻炎性级联反应,保护神经元,发挥其防治脑缺血再灌注损伤的作用。Objective： To investigate the influence of Qingre Huayu FormulaⅡ（QRHYⅡ） on expression of toll like receptor-3（TLR3） and TRIF related adaptor molecule（TRIFM） in rats with cerebral ischemia reperfusion injury and discuss the mechanism of nerve protective effect of QRHYⅡ on rats with cerebral ischemia reperfusion injury. Methods： One hundred and ninety-two healthy SD rats were randomly divided into two groups, and then the two groups were divided in to four subgroups as control group（group A）, sham operation group（group B）, cerebral ischemia-reperfusion group（group C） and QRHYⅡgroup（group D）. Secondary suture occluded method was used to establish the rat models of focal cerebral ischemia reperfusion. The specimen in group B, C and D were collected on different phase accordingly. The specimen were detected by using HE staining and observed with optical microscopes. The expression of TLR3 m RNA and TRAM m RNA on hippocampus of the ischemic side were measured by using fluorescence quantitative PCR. Results： HE staining： Neuron injury could not found in group A and group B under the optical microscope, while there were different degrees of neuron injury in group C and D, and the number of damaged neurons in group D were less than that of group C at each time point. Real time PCR： The expression of TLR3 m RNA and TRAM m RNA in group A and B was in a low level, and compared with group B, the expression of TLR3 m RNA and TRAM m RNA in group C was increased obviously, and the difference between the two groups was statistically significant（P〈0.05）. The expression of TLR3 m RNA and TRAM m RNA began to increase at 3h, and reached its peak by 24 h, while it declined gradually after 48 h in model group. The expression of TLR3 m RNA and TRAM m RNA in group D was lower significantly than that of group C at each time point, and the difference between the two groups was significant（P〈0.05）. Conclusion： QRHYⅡ playes a role in protecting neuron, relieving the inflammatory c

关 键 词：清热化瘀Ⅱ号方 脑缺血再灌注 TOLL样受体3 TRIF相关受体衔接分子 

分 类 号：R285.5[医药卫生—中药学]