摘要
目的:探讨银杏叶片对砷中毒大鼠肺损伤的拮抗与治疗作用及其机制。方法:共选择42只健康清洁级Wistar大鼠,雌雄各半,体重为120~130 g,按体重采用随机数字表法分为7组,每组6只。建立银杏叶片拮抗和银杏叶片治疗两种干预模型,具体处理如下:(1)银杏叶片拮抗实验研究(4组):对照A组给予去离子水;银杏叶片对照组(GBE组)给予银杏叶片溶液(50 mg·kg^(-1)·bw);砷中毒组(As组)给予亚砷酸钠(NaAsO_(2))溶液(10 mg·kg^(-1)·bw);银杏叶片拮抗组(As+GBE组)给予NaAsO_(2)溶液(10 mg·kg^(-1)·bw)处理的同时给予银杏叶片溶液(50 mg·kg^(-1)·bw),以上给药方式均为灌胃,每周6天,连续4个月。(2)银杏叶片治疗实验研究(3组):对照B组给予去离子水5.5个月;砷中毒自然恢复组(恢复组)给予NaAsO_(2)溶液(10 mg·kg^(-1)·bw)灌胃4.0个月后,用去离子水灌胃1.5个月;银杏叶片治疗组(治疗组)给予NaAsO_(2)溶液(10 mg·kg^(-1)·bw)灌胃4.0个月后,给予银杏叶片溶液(50 mg·kg^(-1)·bw)灌胃1.5个月,以上给药均为每周6天。采用Masson染色评价肺组织胶原纤维沉积情况;蛋白免疫印迹法(Western blot)检测肺组织匀浆中相关蛋白[炎性细胞因子基质金属蛋白酶(MMP)-9、白细胞介素(IL)-1β、IL-18,高迁移率族蛋白B1(HMGB1)/晚期糖基化终末产物受体(RAGE)通路的HMGB1、RAGE,磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)通路的PI3K、AKT、磷酸化AKT(p-AKT),转化生长因子(TGF)-β1/SMAD通路的TGF-β1、SMAD2、p-SMAD2、SMAD3、p-SMAD3、SMAD4]的表达水平。结果:(1)银杏叶片拮抗作用:与对照A组比较,GBE组大鼠肺组织各蛋白表达水平和胶原纤维沉积均未见明显变化(P均>0.05),As组大鼠肺组织MMP-9、IL-1β、IL-18蛋白表达水平和胶原纤维沉积均明显增加(P均<0.05),HMGB1、RAGE、PI3K、p-AKT、TGF-β1、p-SMAD2、p-SMAD3、SMAD4蛋白表达水平均明显升高(P均<0.05);与As组比较,As+GBE组大鼠肺组织MMP-9、IL-1β、IL-18蛋白表达水平和胶原纤维沉积均明显降低(P均<0.05),HMGB1、RAGE、PI3K、p-AKT、TGF-β1、p-SMAD2、p-SMAD3蛋白表达水平均明显下降(P均<0.05)。(2)银杏叶片治疗作用:与对照B组比较,恢复组大鼠肺组织MMP-9、IL-1β、IL-18蛋白表达水平和胶原纤维沉积明显增加(P均<0.05),HMGB1、RAGE、PI3K、p-AKT、TGF-β1、p-SMAD2、p-SMAD3、SMAD4蛋白表达水平均明显升高(P均<0.05);与恢复组比较,治疗组大鼠肺组织MMP-9、IL-1β、IL-18、HMGB1、RAGE、PI3K、p-AKT蛋白表达水平均明显降低(P均<0.05),肺组织胶原纤维沉积及TGF-β1、p-SMAD2、p-SMAD3、SMAD4蛋白表达水平均未见明显变化(P均>0.05)。两种实验中,各组间AKT、SMAD2、SMAD3蛋白表达水平比较,差异均无统计学意义(P均>0.05)。结论:银杏叶片干预可能通过抑制HMGB1/RAGE通路相关蛋白的表达水平,改善砷中毒大鼠肺组织炎性损伤和胶原纤维沉积。
Objective To investigate the antagonistic and therapeutic effects of Ginkgo biloba on arsenic-induced lung injury in rats and its mechanism.Methods A total of 42 healthy clean grade Wistar rats,half male and half female,weighing 120-130 g,were randomly divided into 7 groups with 6 rats in each group.Two intervention models of Ginkgo biloba antagonism and treatment were established,respectively.The specific treatments were as follows:(1)Experimental study on the antagonism of Ginkgo biloba(4 groups):the control A group was given deionized water;the Ginkgo biloba control(GBE)group was given Ginkgo biloba solution(50 mg·kg^(-1)·bw);the arsenic-treated(As)group was given sodium arsenite solution(10 mg·kg^(-1)·bw);the Ginkgo biloba antagonistic(As+GBE)group was treated with sodium arsenite solution(10 mg·kg^(-1)·bw)and Ginkgo biloba solution(50 mg·kg^(-1)·bw),and the above administration was by gavage for 6 days/week,for 4 months.(2)Experimental study on the treatment of Ginkgo biloba(3 groups):the control B group was given deionized water for 5.5 months;in the arsenism natural recovery(recovery)group,sodium arsenite solution(10 mg·kg^(-1)·bw)was administered by gavage for 4.0 months and deionized water for 1.5 months;the Ginkgo biloba treatment(treatment)group was given sodium arsenite solution(10 mg·kg^(-1)·bw)by gavage for 4.0 months and Ginkgo biloba solution(50 mg·kg^(-1)·bw)for 1.5 months,and the above administration was for 6 days/week.Masson staining was used to evaluate collagen fiber deposition in lung tissue.Western blotting was used to detect the expression level of related proteins in lung tissue homogenates,including inflammatory cytokines matrix metalloproteinase-9(MMP-9),interleukin(IL)-1β,IL-18;high mobility group box 1(HMGB1)and receptor for advanced glycation end-products(RAGE)of the HMGB1/RAGE pathway;phosphatidylinositol-4,5-bisphosphate 3-kinase(PI3K),protein kinase B(AKT),phosphorylated AKT(p-AKT)of the PI3K/AKT pathway;transforming growth factor(TGF)-β1,SMAD2,p-SMAD2,SMAD3,p-SMAD3 and SMAD4 of the TGF-β1/SMAD pathway.Results(1)Antagonistic effect of Ginkgo biloba:compared with the control A group,there was no significant change in protein expression and collagen fiber deposition in the lung tissue of GBE group(P>0.05);the levels of MMP-9,IL-1βand IL-18 protein expression and collagen fiber deposition in the lung tissue of As group were significantly increased(P<0.05);and the levels of HMGB1,RAGE,PI3K,p-AKT,TGF-β1,p-SMAD2,p-SMAD3 and SMAD4 protein expression were significantly increased(P<0.05).Compared with As group,the levels of MMP-9,IL-1βand IL-18 protein expression and collagen fiber deposition were significantly decreased in As+GBE group(P<0.05);and levels of HMGB1,RAGE,PI3K,p-AKT,TGF-β1,p-SMAD2,and p-SMAD3 protein expression were significantly decreased(P<0.05).(2)Therapeutic effect of Ginkgo biloba:compared with control B group,the levels of MMP-9,IL-1β,IL-18 protein expression and collagen fiber deposition were significantly increased in recovery group(P<0.05);and the levels of HMGB1,RAGE,PI3K,p-AKT,TGF-β1,p-SMAD2,p-SMAD3 and SMAD4 protein expression were significantly increased(P<0.05).Compared with recovery group,the levels of MMP-9,IL-1β,IL-18,HMGB1,RAGE,PI3K and p-AKT protein expression were significantly decreased in treatment group(P<0.05);and there was no significant change in collagen fiber deposition and TGF-β1,p-SMAD2,p-SMAD3 and SMAD4 protein expression levels in lung tissue(P>0.05).In both experiments,there was no significant difference in the protein expression levels of AKT,SMAD2 and SMAD3 between the groups(P>0.05).Conclusion Ginkgo biloba intervention has ameliorated inflammatory injury and collagen fiber deposition in lung tissue of arsenic-treated rats possibly by inhibiting the expression levels of HMGB1/RAGE pathway-related proteins.
作者
郑凡艳
王文娟
张玉琼
张爱华
Zheng Fanyan;Wang Wenjuan;Zhang Yuqiong;Zhang Aihua(Department of Toxicology,Key Laboratory of Environmental Pollution and Disease Control,Ministry of Education,School of Public Health,Guizhou Medical University,Guiyang 550025;Guizhou Center for Disease Control and Prevention,Guiyang 550004,China)
出处
《中华地方病学杂志》
CAS
北大核心
2022年第7期524-531,共8页
Chinese Journal of Endemiology
基金
国家自然科学基金(81430077、U1812403、81872569)。
关键词
砷中毒
银杏叶片
肺
炎症
胶原纤维沉积
Arsenic poisoning
Ginkgo biloba
Lung
Inflammation
Collagen fiber deposition
