化浊解毒活血通络方对脑缺血再灌注损伤大鼠氧化损伤和炎症因子的影响

Effects of Huazhuo Jiedu Huoxue Tongluo Prescription on oxidative injury and inflammatory factors in rats with cerebral ischemia-reperfusion injury

目的:观察化浊解毒活血通络方对脑缺血再灌注损伤大鼠的保护作用及其抗氧化、抗炎性损伤的作用机制。方法:将50只雄性SD大鼠随机分为假手术组,模型组,中药高、中、低剂量组,每组10只。采用改良线栓法制备大脑中动脉阻塞/再灌注大鼠模型,造模24 h后,中药高、中、低剂量组给予相应浓度化浊解毒活血通络方灌胃,连续给药14 d,假手术组和模型组灌胃等体积蒸馏水。观察各组大鼠一般情况,记录大鼠体质量;评定神经功能缺损程度;HE染色观察脑组织结构变化;免疫荧光检测梗死侧脑组织活性氧(ROS)含量;比色法检测大鼠血清中丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性;ELISA法检测血清白细胞介素(IL)-1β、IL-18含量。结果:与假手术组比较,模型组大鼠一般情况较差,取材前体质量显著下降(P<0.05),神经功能缺损评分显著升高(P<0.05),梗死侧脑组织有大量细胞变形、坏死、液化,伴有炎症细胞浸润,脑组织ROS荧光范围和强度明显增加,血清MDA、IL-1β和IL-18含量均显著升高(P<0.05),SOD活性显著降低(P<0.05)。与模型组比较,各给药组大鼠一般情况有所改善,取材前体质量显著增加(P<0.05),中药高剂量组神经功能缺损评分显著下降(P<0.05),各给药组脑组织病理状态有所改善,脑组织ROS荧光范围均有所减少,荧光强度均有不同程度的减弱,各给药组血清MDA含量均显著下降(P<0.05),中药高、中剂量组SOD活性显著提升(P<0.05),血清IL-1β和IL-18含量均显著下降(P<0.05)。结论:化浊解毒活血通络方可能通过减少大鼠脑组织ROS含量和血清MDA含量,增加SOD活性,减少脑组织炎症细胞浸润和炎症因子IL-1β、IL-18的表达而发挥脑保护作用,且药物剂量与疗效总体呈现量效关系。

Objective:To observe the protective effect of Huazhuo Jiedu Huoxue Tongluo Prescription on cerebral ischemia-reperfusion injury in rats and its mechanism of antioxidation and anti-inflammatory injury.Methods:Fifty male SD rats were randomly divided into sham surgery group,model group,and high-dose,medium-dose,and low-dose traditional Chinese medicine groups,with 10 rats in each group.A rat model of middle cerebral artery occlusion/reperfusion was prepared using an improved thread occlusion method.After 24 hours of modeling,the high,medium,and low dose groups of traditional Chinese medicine were given corresponding concentrations of Huazhuo Jiedu Huoxue Tongluo Prescription by gavage for 14 consecutive days.The sham surgery group and model group were given equal volume distilled water by gavage.The general situation of each group of rats was observed and their body weight was recorded;The degree of neurological deficits was assessed;HE staining was used to observe changes in brain tissue structure;Immunofluorescence was used to detect reactive oxygen species(ROS)content in infarcted brain tissue;Colorimetric method was used to detect the content of malondialdehyde(MDA)and the activity of superoxide dismutase(SOD)in rat serum;ELISA method was used to detect the levels of serum interleukin(IL)-1βand IL-18.Results:Compared with the sham surgery group,the general condition of the model group rats was worse,with a significant decrease in precursor mass(P<0.05),a significant increase in neurological deficit score(P<0.05),and a large amount of cell deformation,necrosis,and liquefaction in the infarcted brain tissue,accompanied by inflammatory cell infiltration.The ROS fluorescence range and intensity in the brain tissue were significantly increased,and serum MDA,IL-1β,and IL-18 levels were significantly increased(P<0.05),while SOD activity was significantly reduced(P<0.05).Compared with the model group,the general condition of rats in each treatment group was improved,with a significant increase in the mass of precursor samples(P<0.05).The neurological deficit score in the high-dose group of traditional Chinese medicine was significantly reduced(P<0.05),and the pathological state of brain tissue in each treatment group was improved.The ROS fluorescence range of brain tissue was also reduced,and the fluorescence intensity was weakened to varying degrees.The serum MDA content in each treatment group was significantly reduced(P<0.05).The SOD activity in the high-and medium-dose group was significantly increased(P<0.05),while the serum levels of IL-1βand IL-18 were significantly decreased(P<0.05).Conclusion:The Huazhuo Jiedu Huoxue Tongluo Prescription may exert neuroprotective effects by reducing the ROS content in rat brain tissue and serum MDA content,increasing SOD activity,reducing the infiltration of inflammatory cells in brain tissue,and the expression of inflammatory factors IL-1βand IL-18.The overall dose-response relationship between drug dosage and therapeutic effect is dose-response.