三白草酮对UUO小鼠肾组织纤维化的抑制作用及其机制  

Inhibitory effect and mechanism of Sauchinone on renal fibrosis in unilateral ureteral obstruction mice

作　　者：吴朋烊 赵铃 李秋月 郭红艳[1] 李超君[1] 吴丽娜[1] 赵正林 WU Pengyang;ZHAO Ling;LI Qiuyue;GUO Hongyan;LI Chaojun;WU Lina;ZHAO Zhenglin(School of Medical Technology,Qiqihar Medical University,Qiqihar 161006,China;Yanbian University College of Medicine,Yanji 133002,China)

机构地区：[1]齐齐哈尔医学院医学技术学院,黑龙江齐齐哈尔161006 [2]延边大学医学院,吉林延吉133002

出　　处：《中国实验动物学报》2024年第10期1270-1280,共11页Acta Laboratorium Animalis Scientia Sinica

基　　金：黑龙江省卫生健康委科研课题(20210202020397);黑龙江省省属高等学校基本科研业务费科研项目(2022-KYYWF-0830);齐齐哈尔医学院研究生创新基金项目(QYYCX2023-02)。

摘　　要：目的探究三白草酮(Sauchinone,Sch)抑制单侧输尿管结扎(unilateral ureteral obstruction,UUO)小鼠肾组织纤维化作用及其机制。方法取雄性ICR小鼠40只,分为假手术组(Sham group)、UUO纤维化模型组(UUO-Model group)、Sch低剂量组(Sch-Low group)、Sch高剂量组(Sch-High group)以及阳性对照组(Val group)。假手术组仅分离左侧肾输尿管,不结扎。UUO纤维化模型组剥离左侧近端输尿管进行双结扎,手术后第2天开始,Sch低剂量组(10 mg/kg)、Sch高剂量组(30 mg/kg)、阳性对照组(缬沙坦100 mg/kg)分别每日灌胃1次,连续灌胃给药4周。给药结束后,检测血清中肌酐(serum creatinine,SCr)、血尿素氮(blood urea nitrogen,BUN)以及炎症因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)水平;检测肾组织中超氧化物歧化酶(superoxide dismutase,SOD)活性、丙二醛(malondialdehyd,MDA)和活性氧(reactive oxygen species,ROS)含量;实时荧光定量逆转录PCR(reverse transcription real-time quantitative polymerase chain reaction,RT-qPCR)法检测肾组织中炎症因子TNF-α、IL-6 mRNA水平;采用苏木素-伊红(HE)、马松(Masson)染色、免疫组化法检测小鼠肾组织病理变化和胶原沉积以及转化生长因子β1(transforming growth factor-β1,TGF-β1)、SMAD家族成员3(SMAD family member 3,SMAD3)和结缔组织生长因子(connective tissue growth factor,CTGF)蛋白表达情况;蛋白质免疫印记法(Western Blot)观察肾组织中胶原蛋白Ⅰ型(collagen typeⅠ,CollagenⅠ)、CTGF、SMAD3、核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)、血红素氧合酶-1(heme oxygenase-1,HO-1)、核因子-κB(nuclear factor kappa-B,NF-κB)蛋白表达。结果与UUO纤维化模型组比较,Sch低剂量组和Sch高剂量组小鼠血清中SCr和BUN以及TNF-α、IL-6水平显著降低(P<0.05或P<0.01);肾组织中SOD活性显著升高且MDA和ROS含量显著降低(P<0.05或P<0.01);肾组织中TNF-α、IL-6 mRNA的Objective To investigate the inhibitory effect of Sauchinone(Sch)on renal tissue fibrosis and to explore its mechanism in unilateral ureteral obstruction(UUO)mice.Methods Forty male ICR mice were divided into Sham,UUO-Model,Sch Low-dose,Sch High-dose,and positive control(Val)groups.Mice in the sham group underwent isolation of the left renal ureter without ligation,and mice in the UUO-Model group underwent stripping of the left proximal ureter for double ligation.Starting on day 2 after surgery,the Sch Low-dose group received 10 mg/kg Sch,the Sch High dose group received 30 mg/kg Sch,and the Val group received 100 mg/kg valsartan by gavage once daily for 4 weeks.At the end of drug administration,serum creatinine(SCr),blood urea nitrogen(BUN),and the inflammatory factors tumor necrosis factor(TNF)-αand interleukin(IL)-6 were measured.Superoxide dismutase(SOD)activity,and malondialdehyde(MDA)and reactive oxygen species(ROS)content were detected in renal tissues.TNF-αand IL-6 mRNA levels were detected in renal tissues by reverse transcription-quantitative polymerase chain reaction.Pathological changes and collagen deposition,as well as transforming growth factor(TGF)-β1,Smad3,and connective tissue growth factor(CTGF)protein expression in mouse renal tissues were detected by hematoxylin and eosin and Masson staining,and immunohistochemistry.CollagenⅠ,CTGF,Smad3,nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1),and nuclear factor(NF)-κB protein expression in renal tissues were detected by Western Blot.Results SCr,BUN,TNF-α,and IL-6 were all significantly reduced in Sch Low-dose and Sch High-dose mice compared with the UUO Model group(P<0.05 or P<0.01),while SOD activity was significantly higher and MDA and ROS levels were significantly lower in renal tissues(P<0.05 or P<0.01),and TNF-αand IL-6 mRNA in renal tissues were significantly reduced(P<0.05 or P<0.01).Microscopic observation of swollen renal tubules with thylakoid hyperplasia and collagen fiber deposition was significantly improv

关 键 词：三白草酮 肾纤维化 CTGF TGF-Β1 氧化应激 炎症因子 

分 类 号：Q95-33[生物学—动物学]