摘要
目的 探究银杏达莫预处理对大鼠小肠缺血-再灌注损伤的影响及相关机制。方法 采用结扎肠系膜上动脉模拟大鼠小肠缺血-再灌注损伤模型,根据随机数字表法将36只Wistar大鼠随机分为假手术组(Sham组),小肠缺血-再灌注模型组(I/R组),银杏达莫干预组(Gin组),每组12只。建模前7d Gin组按3.6mL/1kg给药量进行腹腔注射,一天一次,连续7d, Sham组、I/R组注射等剂量生理盐水。苏木素-伊红(HE)染色观察细胞状态,天狼星红染色和Masson染色观察胶原纤维;血液生化检测血清中超氧化物歧化酶(SOD)和丙二醛(MDA)含量变化;免疫组化检测小肠组织中白介素6(IL-6)、白介素10(IL-10)、Bcl-相关X蛋白(Bax)、B细胞淋巴瘤/白血病-2(Bcl-2)的蛋白表达变化;聚合酶链式反应(PCR)检测Bax、Bcl-2的mRNA表达水平;免疫印迹法(Western blot)检测小肠组织中Smad蛋白(Smad)和Bcl-2,Bax的蛋白表达水平。结果 与Sham组相比,I/R组小肠组织中SOD活性[(125.34±59.58)U/L]降低(P<0.05),MDA含量[(6.24±0.73)U/L]增加(P<0.05);与I/R组相比,Gin组小肠组织中SOD活性[(262.62±22.57)U/L]增强(P<0.05),MDA含量[(2.88±0.74)U/L]降低(P<0.05)。Sham组小肠黏膜结构正常,绒毛完整,I/R组可见大量裸露绒毛,绒毛出现水肿,脱落受损严重,并且有炎性细胞浸润现象,Gin组小肠绒毛结构破坏较轻,可见黏膜轻微水肿和轻微脱落,损伤较轻;天狼星红和Masson染色发现I/R组中胶原纤维较Sham组明显增多,Gin组中的胶原纤维较I/R组有所减轻;免疫组化结果显示,I/R组IL-6、IL-10、Bax、Bcl-2阳性面积占比[(32.24±7.07)%、(10.23±1.63)%、(40.35±7.07)%、(24.64±8.04)%]均高于Sham组[(4.24±1.31)%、(3.77±0.91)%、(2.35±0.87)%、(3.25±0.87)%](P<0.05),Gin组IL-6、Bax阳性面积[(10.23±0.71)%、(6.71±1.41)%]低于I/R组(P<0.05),IL-10、Bcl-2阳性面积[(32.24±8.16)%、(48.40±7.07)%]均高于I/R组(P<0.05);与Sham组相比,I/R组大鼠小肠组织中Bax、Bcl-2、Akt、Smad mRNA表达水平均上调(P<0.05),与I/R组比较,Gin组Bax和Smad mRNA表达水平降低(P<0.05),Bcl-2和Akt蛋白相应的mRNA表达水平上调(P<0.05);与Sham组相比,I/R组大鼠小肠组织中Akt、Smad、Bax、Bcl-2蛋白表达水平上调(P<0.05)。与I/R组比较,Gin组大鼠小肠组织中Akt和Bcl-2蛋白表达水平显著上调(P<0.05),Smad和Bax蛋白表达水平显著降低(P<0.05)。结论 银杏达莫通过调节抗氧化酶活性、减轻炎症反应以及抑制细胞凋亡相关蛋白的表达对大鼠小肠缺血-再灌注损伤有保护作用。
Objective To explore the effect of Ginkgodamol preconditioning on small intestine ischemia-reperfu-sion injury in rats and its related mechanism.Methods A rat small intestine ischemia-reperfusion injury model was simulated by ligating superior mesenteric artery.According to the random number table,36 Wistar rats were randomly divided into normal group(Sham group),small intestine ischemia-reperfusion model group(I/R group)and Ginkgo-damol intervention group(Gin group),with 12 rats in each group.Seven days before modeling,the Ginkgodamol(Gin)group was injected intraperitoneally at a dosage of 3.6mL/1000g once a day for seven days,while the Sham group and I/R group were injected with the same dose of normal saline.HEmatoxylin-eosin(HE)staining was used to observe the cell state,while Sirius red staining and Masson staining were used to observe the collagen fibers.The contents of superoxide dismutase(SOD)and malondialdehyde(MDA)in serum were detected by blood biochemistry.Immunohisto-chemistry was used to detect the protein expression changes of interleukin-6(IL-6),interleukin-10(IL-10),Bcl-related X protein(Bax)and B-cell lymphoma/leukemia-2(Bcl-2)in small intestine.The mRNA expression levels of Bax and Bcl-2 were detected by polymerase chain reaction(PCR).Western blot was used to detect the protein expression levels of Smad,Bcl-2 and Bax in small intestine.Results Compared with Sham group,the SOD activity(125.34±59.58)u/L in I/R group decreased(P<0.05),and the MDA content(6.24±0.73)u/L increased(P<0.05).Compared with I/R group,SOD activity(262.62±22.57)u/L in small intestine of Gin group was increased(P<0.05),while MDA content(2.88±0.74)u/L was decreased(P<0.05).The results of HE staining showed that the intestinal mucosal structure in Sham group was normal,and the villi were intact.In I/R group,a large number of exposed villi were seen,and the villi were edema,shedding and damaged seriously,and inflammatory cells infiltrated.In Gin group,the structural damage of intestinal villi was light,and the mucosal edema and shedding were slight,and the damage was light.Sirius red and Masson staining showed that collagen fibers in I/R group were significantly more than those in Sham group,and collagen fibers in Gin group were less than those in I/R group.Immunohistochemical results showed that the positive areas of IL-6,IL-10,Bax and Bcl-2 in I/R group[(32.24±7.07)%,(10.23±1.63)%,(40.35±7.07)%and(24.64±8.04)%]were higher than those in Sham group[(4.24±1.31)%,(3.77±0.91)%,(2.35±0.87)%,(3.25±0.87)%](P<0.05).The positive areas of IL-6 and Bax in Gin group[(10.23±0.71)%,(6.71±1.41)%]were lower than those in I/R group.The results of PCR showed that compared with Sham group,the expression levels of Bax,Bcl-2,Akt and Smad mRNA in small intestine of rats in I/R group were all up-regulated(P<0.05),while compared with I/R group,the expression levels of Bax and Smad mRNA in Gin group were down-regulated(P<0.05),and the correspond-ing mRNA expression levels of Bcl-2 and Akt protein were up-regulated(P<0.05).Western blot showed that the expres-sion levels of Akt,Smad,Bax and Bcl-2 protein in small intestine of rats in I/R group were up-regulated compared with Sham group(P<0.05).Compared with I/R group,the expression levels of Akt and Bcl-2 protein in small intestine of rats in Gin group were significantly increased(P<0.05),while the expression levels of Smad and Bax protein were significantly decreased(P<0.05).Conclusion Ginkgodamol has protective effect on ischemia-reperfusion injury of small intestine in rats by regulating antioxidant enzyme activity,reducing inflammatory responses,and inhibiting the expression of apoptosis-related proteins.
作者
刘白露
刘梓育
李研思
聂瑜
张璟玥
刘志强
LIU Bailu;LIU Ziyu;LI Yansi;NIE Yu;ZHANG Jingyue;LIU Zhiqiang(School of Pharmacy,Jining Medical University,Rizhao 276826,China;School of Pharmacy,Yantai University,Collaborative Innovation Center of Advanced Drug Delivery System and Biotech Drugs of Shandong Universities,Yantai 264005,China;School of Nursing,Jining Medical University,Jining 272000,China)
出处
《济宁医学院学报》
2025年第1期19-26,共8页
Journal of Jining Medical University
基金
cx2021134济宁医学院大学生创新训练计划项目(2021-ZXJK-08山东省人文社会科学课题)。
作者简介
通信作者:刘志强,Email:liuzhiqiangjy@yeah.net。
