虎杖预处理对大鼠肾缺血再灌注损伤的保护作用
摘要
目的：探讨虎杖预处理对肾缺血/再灌注(Ischemia-reperfusion, I/R)损伤大鼠肾脏的保护作用及其机制。方法：健康雄性SD大鼠72只随机分为四组,假手术组、I/R组、虎杖预处理及虎杖苷(PD)预处理组每组18只,每组根据再灌注时间不同分为三个时间点(缺血前10min、再灌注后6、12h),每个时间点随机取6只大鼠。采用切除大鼠右侧肾脏,夹闭左侧肾蒂60min的肾缺血再灌注模型。分别检测各组大鼠血清肌酐(Scr)、尿素氮(BUN)、肾组织丙二醛(MDA)水平及超氧化物歧化酶(SOD)活性,观察肾组织的病理变化,免疫组化法检测肾组织NF-κBp65、ICAM-1、ET的表达,ELISA法检测血清中TNF-a的含量,并分别进行比较。结果：1.造模组可见肾小管扩张,肾小管上皮细胞肿胀,并有不同程度的变性、坏死,部分细胞核溶解,部分肾小管可见空泡样变性,再灌注12h后较6h后更为明显,治疗组这些病理变化明显减轻。2.再灌注6h、12h后I/R组、虎杖预处理及PD预处理组Scr、BUN、肾组织MDA、血清TNF-α水平及肾组织NF-κB、ICAM-1、ET阳性表达均较假手术组升高(P<0.05),肾组织SOD水平较假手术组降低(P<0.05)。术前10min时无明显变化。3.再灌注6h、12h后虎杖预处理及PD预处理组Scr、BUN、肾组织MDA水平及肾组织NF-κB、ICAM-1阳性表达均较I/R组降低(P<0.05),术前10min各组间无明显变化。4.再灌注12h后虎杖预处理及PD预处理组SOD水平较I/R组升高(P<0.05),再灌注6h后PD预处理组较I/R组无明显变化。5.再灌注6h、12h后虎杖预处理组肾组织ET阳性表达低于I/R组(P<0.05),PD预处理组无明显变化。6.再灌注6h后虎杖预处理及PD预处理组血清TNF-α含量明显低于I/R组(P<0.05),再灌注12h后无明显变化。结论：在肾缺血再灌注损伤的过程中,虎杖可能通过增强SOD活性,减少MDA、ICAM-1、NF-κBp65、ET、TNF-α的水平,改善肾功能的作用来保护肾脏,其中PD起了主要作用。
Objective:To investigate the protective effect and mechanisms of polygonum on renal ischemia-reperfusion injury in rats. Methods:72 SD male rats were randomly divided into four groups:sham-operation group, ischemia-reperfusion group, polygonum cuspidatum preconditioning group and polydatin (PD) preconditioning group.Every group was divided into three groups in different time points(Before ischemia 10min, after ischemia-reperfusion 6,12 h).Six rats were conducted to experiment in each time point. The rat's RIRI model was established by resecting the right renal firstly, followed by blocking the blood flow of left renal for 60 min. Renal function was assessed by measuring serum creatinine and blood urea nitrogen.The MDA content were measured by Thibabituric Acid technique and SOD activity were measured xanthine oxidase technique. The protein expression of NF-κB, ICAM-1 and ET was measured immunohistochemistry. ELISA assay of renal tissue levels of TNF-a, and were compared. Results:1. The model group were found epithelial cells edema of renal tubule, vacuole-zation,tuble dilation, shrinkage of basement membranes,nuclear fragmentation and various tuble casta,while all these histopathological changes were relieved in treatment group。2. I/R group、polygonum cuspidatum group and PDgroup showed significant increase in Scr、BUN、MDA、TNF-、the expression of NF-κB、ICAM-1、ET(P<0.05)and decrease in SOD compared with sham-operation group at 6、12 hours after renal reperfusion. All preoperativelOmin groups have no difference with ham-operation group.3. Polygonum cuspidatum preconditioning group and PD preconditioning group showed significant decrease in Scr、BUN、MDA、TNF-α、the expression of NF-κB、ICAM-1 compared with the I/R group at 6、12 hours after renal reperfusion (P<0.05).4. Polygonum cuspidatum preconditioning group and PD preconditioning group showed increase in SOD compared with I/R group at 12 hours after renal reperfusion(P<0.05). PD preconditioning group have no difference with the ischemia-reperfusion group at 6 hours after renal reperfusion.5. Polygonum cuspidatum preconditioning group showed significant decrease in the expression of ET compared with the I/R group at 6、12 hours after renal reperfusion(P<0.05). PD preconditioning group have no difference with the I/R group.6. Polygonum cuspidatum preconditioning group and PD preconditioning group showed significant decrease in TNF-a compared with I/R group at 6 hours after renal reperfusion(P<0.05),and have no difference with each other at 6 hours after renal reperfusion. Conclusions:In the process of renal I/R injury polygonum cuspidatum can enhance SOD activity, reduced MDA, ICAM-1, NF-κBp65, ET, to improve renal function, which played a major role in PD but there are other ingredients play a role.
Objective:To investigate the protective effect and mechanisms of polygonum on renal ischemia-reperfusion injury in rats. Methods:72 SD male rats were randomly divided into four groups:sham-operation group, ischemia-reperfusion group, polygonum cuspidatum preconditioning group and polydatin (PD) preconditioning group.Every group was divided into three groups in different time points(Before ischemia 10min, after ischemia-reperfusion 6,12 h).Six rats were conducted to experiment in each time point. The rat's RIRI model was established by resecting the right renal firstly, followed by blocking the blood flow of left renal for 60 min. Renal function was assessed by measuring serum creatinine and blood urea nitrogen.The MDA content were measured by Thibabituric Acid technique and SOD activity were measured xanthine oxidase technique. The protein expression of NF-κB, ICAM-1 and ET was measured immunohistochemistry. ELISA assay of renal tissue levels of TNF-a, and were compared. Results:1. The model group were found epithelial cells edema of renal tubule, vacuole-zation,tuble dilation, shrinkage of basement membranes,nuclear fragmentation and various tuble casta,while all these histopathological changes were relieved in treatment group。2. I/R group、polygonum cuspidatum group and PDgroup showed significant increase in Scr、BUN、MDA、TNF-、the expression of NF-κB、ICAM-1、ET(P<0.05)and decrease in SOD compared with sham-operation group at 6、12 hours after renal reperfusion. All preoperativelOmin groups have no difference with ham-operation group.3. Polygonum cuspidatum preconditioning group and PD preconditioning group showed significant decrease in Scr、BUN、MDA、TNF-α、the expression of NF-κB、ICAM-1 compared with the I/R group at 6、12 hours after renal reperfusion (P<0.05).4. Polygonum cuspidatum preconditioning group and PD preconditioning group showed increase in SOD compared with I/R group at 12 hours after renal reperfusion(P<0.05). PD preconditioning group have no difference with the ischemia-reperfusion group at 6 hours after renal reperfusion.5. Polygonum cuspidatum preconditioning group showed significant decrease in the expression of ET compared with the I/R group at 6、12 hours after renal reperfusion(P<0.05). PD preconditioning group have no difference with the I/R group.6. Polygonum cuspidatum preconditioning group and PD preconditioning group showed significant decrease in TNF-a compared with I/R group at 6 hours after renal reperfusion(P<0.05),and have no difference with each other at 6 hours after renal reperfusion. Conclusions:In the process of renal I/R injury polygonum cuspidatum can enhance SOD activity, reduced MDA, ICAM-1, NF-κBp65, ET, to improve renal function, which played a major role in PD but there are other ingredients play a role.
引文
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