CAJ | 학술논문

目的探讨青蒿琥酯对大鼠肾脏缺血再灌注后纤维化改变的保护作用及其机制.方法将18只大鼠随机分为3组,每组6只.假手术组:戊巴比妥钠麻醉(40 mg/kg)后,取腹正中切口进入腹腔,游离双侧肾脏,切除右肾后缝合腹壁;缺血再灌注组:在切除右肾后,用无创伤血管夹夹闭左肾动、静脉45 min进行缺血,然后再灌注8周,余同假手术组;青蒿琥酯组:再灌注24h后,开始青蒿琥酯(每天25 mg/kg)灌胃处理,假手术组和缺血再灌注组等体积蒸馏水灌胃,余同缺血再灌注组.再灌注8周后,检测各组大鼠血清肌酐、尿素氮水平,苏木素-伊红(HE)染色、Masson染色观察肾组织结构变化和纤维化改变,免疫组织化学观察纤维连接蛋白(Fibronectin)表达,实时荧光定量聚合酶链反应(Real-time PCR)检测α-平滑肌肌动蛋白(α-SMA)水平,Western blot检测结缔组织生长因子(CTGF)和转化生长因子-β1(TGF-β1)的表达水平.结果在假手术组、缺血再灌注组和青蒿琥酯组中,血清肌酐(μmoL/L)分别为30.13±1.52、32.36±2.67、31.29±1.36,血清尿素氮(mmol/L)分别为6.86±0.79、7.43±1.42、7.27 ±0.61,假手术组的血清肌酐、尿素氮水平明显比缺血再灌注组和青蒿琥酯组低(P＜0.05),而缺血再灌注组的血清肌酐、尿素氮水平显著高于青蒿琥酯组(P＜0.05).HE染色可见缺血再灌注组中的肾小管上皮细胞明显变性坏死,有大量炎性细胞浸润,而青蒿琥酯可以明显减轻损伤.Masson染色结果可见缺血再灌注组纤维化改变明显,而青蒿琥酯可以明显减轻纤维化改变.与假手术组比较,缺血再灌注组和青蒿琥酯组Fibronectin、α-SMA、CTGF和TGF-β1水平均显著升高,而青蒿琥酯组的上述指标明显低于缺血再灌注组.结论青蒿琥酯可以减轻大鼠肾脏缺血再灌注损伤导致的纤维化改变,其机制可能与抑制相关蛋白的表达有关.
목적 탐토청호호지대대서신장결혈재관주후섬유화개변적보호작용급기궤제.방법 장18지대서수궤분위3조,매조6지.가수술조:무파비타납마취(40 mg/kg)후,취복정중절구진입복강,유리쌍측신장,절제우신후봉합복벽;결혈재관주조:재절제우신후,용무창상혈관협협폐좌신동、정맥45 min진행결혈,연후재관주8주,여동가수술조;청호호지조:재관주24h후,개시청호호지(매천25 mg/kg)관위처리,가수술조화결혈재관주조등체적증류수관위,여동결혈재관주조.재관주8주후,검측각조대서혈청기항、뇨소담수평,소목소-이홍(HE)염색、Masson염색관찰신조직결구변화화섬유화개변,면역조직화학관찰섬유련접단백(Fibronectin)표체,실시형광정량취합매련반응(Real-time PCR)검측α-평활기기동단백(α-SMA)수평,Western blot검측결체조직생장인자(CTGF)화전화생장인자-β1(TGF-β1)적표체수평.결과 재가수술조、결혈재관주조화청호호지조중,혈청기항(μmoL/L)분별위30.13±1.52、32.36±2.67、31.29±1.36,혈청뇨소담(mmol/L)분별위6.86±0.79、7.43±1.42、7.27 ±0.61,가수술조적혈청기항、뇨소담수평명현비결혈재관주조화청호호지조저(P＜0.05),이결혈재관주조적혈청기항、뇨소담수평현저고우청호호지조(P＜0.05).HE염색가견결혈재관주조중적신소관상피세포명현변성배사,유대량염성세포침윤,이청호호지가이명현감경손상.Masson염색결과가견결혈재관주조섬유화개변명현,이청호호지가이명현감경섬유화개변.여가수술조비교,결혈재관주조화청호호지조Fibronectin、α-SMA、CTGF화TGF-β1수평균현저승고,이청호호지조적상술지표명현저우결혈재관주조.결론 청호호지가이감경대서신장결혈재관주손상도치적섬유화개변,기궤제가능여억제상관단백적표체유관.
Objective To investigate the effecof artesunate on fibrosiinduced by renal ischemiand reperfusion injury (IRI) in rats.MethodEighteen SD ratwere randomly divided into three groups:sham operated group, IRI group and artesunate group.Each group had 6 rats.In sham operated group, the righkidneyof ratwere excised.In IRI group, lefrenal arterieand veinof ratwere clamped fo45 min, then vasculaclamp waremoved and the ratwere subjected to reperfusion fo8 weeks.In artesunate group, operation stepwere the same athose in IRI group, bu24 h aftereperfusion, artesunate wagiven by intragastriadministration [25 mg/(kg·d)].In sham operated group and IRI group, equal volume of distilled watewagiven.The parameter(blood urenitrogen and creatine) of renal function were determined by auto-biochemieal analyzer.The expression of α-smooth muscle actin (α-SMA) wadetected by real-time quantitative polymerase chain reaction (Real-time PCR) and the changeof renal tissue were examined by hematoxylin and eosin (HE) staining and Masson staining.Immunohistochemistry waperformed to detecfibronectin.The expression of connective tissue growth facto(CTGF) and transforming growth factor-β1 (TGF-β1) wadetected by Western blotting.ResultAmong sham operated group, IRI group and artesunate group, creatine level(μ mol/L) were 30.13 ± 1.52, 32.36 ±2.67 and 31.29 ± 1.36, and blood urenitrogen level(mmol/L) were 6.86 ± 0.79, 7.43 ± 1.42 and 7.27 ± 0.61, respectively.There wano significandifference in renal function between sham operated group and IRI group oartesunate group.HE staining and Masson staining indicated thaartesunate could reduce the injury and fibrosicaused by renal IRI.Acompared with sham operated group, fibronectin, α-SMA,CTGF and TGF-β1 were significantly increased in IRI group and artesunate group.However, these targetin artesunate group were significantly decreased acompared with those in IRI group.Conclusion Artesunate can alleviate fibrosiinduced by renal renal IRI probably by inhibiting the expression of fibronectin, α-SMA, CTGF and TGF-β1.