中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
1期
94-96
,共3页
王李菲%苗志钊%赵筱雯%贺艳%邬善敏
王李菲%苗誌釗%趙篠雯%賀豔%鄔善敏
왕리비%묘지쇠%조소문%하염%오선민
小檗碱%阻塞性黄疸%沉默信息调控因子%超氧化物歧化酶
小檗堿%阻塞性黃疸%沉默信息調控因子%超氧化物歧化酶
소벽감%조새성황달%침묵신식조공인자%초양화물기화매
Berberine%Cholestasis%Silent information regulator 1%Superoxide dismutase
目的 检测肾组织沉默信息调控因子1(SIRT1)蛋白表达和超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、丙二醛(MDA)的水平变化,以及肾细胞凋亡率等变化,探讨小檗碱(BBR)对梗阻性黄疸大鼠肾脏SIRT1蛋白表达的影响及其与肾功能保护的作用.方法 雄性Wistar大鼠45只,随机分为3组,A(Sham)组:假手术组,B(BDL)组:梗阻性黄疸组,C(BDL+BBR)组:梗阻性黄疸+小檗碱干预组.实验结束时取血清检测总胆红素(TB)、直接胆红素(DB)、血尿素氮(BUN)、血肌酐(Cr),肾组织匀浆检测SOD、MDA、GSH,Western blot检测肾组织SIRT1蛋白表达,反转录-聚合酶链反应(RT-PCR)检测肾组织SIRT1 mRNA,原位缺口末端标记法(TUNEL)法检测肾细胞凋亡.结果 B组与A组比较:血清BUN、Cr升高,肾组织SOD、GSH、SIRT1 mRNA及蛋白降低,细胞凋亡率升高(P＜0.05);C组与B组比较:血清BUN、Cr降低,肾组织SOD、GSH、SIRT1 mRNA及蛋白升高,细胞凋亡率降低(P＜0.05).A、B、C组BUN、Cr和肾组织SOD、GSH、SIRT1 mRNA及蛋白、细胞凋亡率指标为:A组(3.14 ±0.53) mmol/L、(31.32±11.08) μmol/L、(19.45 ±2.41) nu/mg、(2.31 ±0.14) nmol/g、1.00±0.00、0.27±0.01、(0.28±0.13)％;B组(8.37±1.57) mmol/L、(102.43±9.32) μmol/L、(5.96±1.43) nu/mg、(1.39±0.25) nmol/g、0.51 ±0.05、0.17±0.01、(19.36±2.56)％;C组(6.04±1.52) mmol/L、(78.58±13.63) μmol/L、(13.52±1.47) nu/mg、(1.97±0.19) nmol/g、0.79 ±0.04、0.22±0.01、(11.47±3.61)％.结论 BBR通过上调SIRT1蛋白促进SOD基因表达,而发挥抗氧化、抗凋亡作用,减轻梗阻性黄疸氧化性肾损害.
目的 檢測腎組織沉默信息調控因子1(SIRT1)蛋白錶達和超氧化物歧化酶(SOD)、穀胱甘肽(GSH)、丙二醛(MDA)的水平變化,以及腎細胞凋亡率等變化,探討小檗堿(BBR)對梗阻性黃疸大鼠腎髒SIRT1蛋白錶達的影響及其與腎功能保護的作用.方法 雄性Wistar大鼠45隻,隨機分為3組,A(Sham)組:假手術組,B(BDL)組:梗阻性黃疸組,C(BDL+BBR)組:梗阻性黃疸+小檗堿榦預組.實驗結束時取血清檢測總膽紅素(TB)、直接膽紅素(DB)、血尿素氮(BUN)、血肌酐(Cr),腎組織勻漿檢測SOD、MDA、GSH,Western blot檢測腎組織SIRT1蛋白錶達,反轉錄-聚閤酶鏈反應(RT-PCR)檢測腎組織SIRT1 mRNA,原位缺口末耑標記法(TUNEL)法檢測腎細胞凋亡.結果 B組與A組比較:血清BUN、Cr升高,腎組織SOD、GSH、SIRT1 mRNA及蛋白降低,細胞凋亡率升高(P＜0.05);C組與B組比較:血清BUN、Cr降低,腎組織SOD、GSH、SIRT1 mRNA及蛋白升高,細胞凋亡率降低(P＜0.05).A、B、C組BUN、Cr和腎組織SOD、GSH、SIRT1 mRNA及蛋白、細胞凋亡率指標為:A組(3.14 ±0.53) mmol/L、(31.32±11.08) μmol/L、(19.45 ±2.41) nu/mg、(2.31 ±0.14) nmol/g、1.00±0.00、0.27±0.01、(0.28±0.13)％;B組(8.37±1.57) mmol/L、(102.43±9.32) μmol/L、(5.96±1.43) nu/mg、(1.39±0.25) nmol/g、0.51 ±0.05、0.17±0.01、(19.36±2.56)％;C組(6.04±1.52) mmol/L、(78.58±13.63) μmol/L、(13.52±1.47) nu/mg、(1.97±0.19) nmol/g、0.79 ±0.04、0.22±0.01、(11.47±3.61)％.結論 BBR通過上調SIRT1蛋白促進SOD基因錶達,而髮揮抗氧化、抗凋亡作用,減輕梗阻性黃疸氧化性腎損害.
목적 검측신조직침묵신식조공인자1(SIRT1)단백표체화초양화물기화매(SOD)、곡광감태(GSH)、병이철(MDA)적수평변화,이급신세포조망솔등변화,탐토소벽감(BBR)대경조성황달대서신장SIRT1단백표체적영향급기여신공능보호적작용.방법 웅성Wistar대서45지,수궤분위3조,A(Sham)조:가수술조,B(BDL)조:경조성황달조,C(BDL+BBR)조:경조성황달+소벽감간예조.실험결속시취혈청검측총담홍소(TB)、직접담홍소(DB)、혈뇨소담(BUN)、혈기항(Cr),신조직균장검측SOD、MDA、GSH,Western blot검측신조직SIRT1단백표체,반전록-취합매련반응(RT-PCR)검측신조직SIRT1 mRNA,원위결구말단표기법(TUNEL)법검측신세포조망.결과 B조여A조비교:혈청BUN、Cr승고,신조직SOD、GSH、SIRT1 mRNA급단백강저,세포조망솔승고(P＜0.05);C조여B조비교:혈청BUN、Cr강저,신조직SOD、GSH、SIRT1 mRNA급단백승고,세포조망솔강저(P＜0.05).A、B、C조BUN、Cr화신조직SOD、GSH、SIRT1 mRNA급단백、세포조망솔지표위:A조(3.14 ±0.53) mmol/L、(31.32±11.08) μmol/L、(19.45 ±2.41) nu/mg、(2.31 ±0.14) nmol/g、1.00±0.00、0.27±0.01、(0.28±0.13)％;B조(8.37±1.57) mmol/L、(102.43±9.32) μmol/L、(5.96±1.43) nu/mg、(1.39±0.25) nmol/g、0.51 ±0.05、0.17±0.01、(19.36±2.56)％;C조(6.04±1.52) mmol/L、(78.58±13.63) μmol/L、(13.52±1.47) nu/mg、(1.97±0.19) nmol/g、0.79 ±0.04、0.22±0.01、(11.47±3.61)％.결론 BBR통과상조SIRT1단백촉진SOD기인표체,이발휘항양화、항조망작용,감경경조성황달양화성신손해.
Objective To determinethe expression of silent information regulator 1 (SIRT1),superoxide dismutase (SOD),malondialdehyde (MDA) and the glutathione (GSH),the aim of this study is to investigate the effects of the berberine to the expression of the SIRT1 protein and the protective effects to the renal function as related to cholestatic kidney injury in rats.Methods The animals were divided randomly into three groupls (n =15).A rat model of cholestasis was established by bile duct ligation (BDL,B group) and compared with a Sham group receiving laparotomy without BDL (Sham,A group),and with the the BBR given respectively following BDL (BDL + BBR,C group).All rats were sacrificed on the 7th day after BDL and the blood and kidney tissue samples were obtained.The expression of the SIRT1 proteins were analyzed by western blotting and the reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to determine the mRNA expression of SIRT1 in all groups.The kits of SOD,MDA and GSH were used to detect the values in renal tissue and the apoptosis of the kidney cells was examined by TdT-mediated dUTP nick end labeling (TUNEL) staining.Results After the bile duct ligation,the model of cholestasis was established.Comparedto A group,the levels of the mRNA and proteins of the SIRT1,SOD and GSH were lower but the alanine transaminase (ALT),MDA and the apoptosis rate were higher in group B (P ＜ 0.05).Compared C groups to B group,the levels of the mRNA and proteins of the SIRT1,SOD and GSH were higher but the MDA and the apoptosis rate were lower (P ＜ 0.05).The values of BUN,Cr,SOD,GSH,SIRT1 mRNA and protein and the rate of the A,B and C groups are:A group (3.14±0.53) mmol/L,(31.32±11.08) μmol/L,(19.45 ±2.41) nu/mg,(2.31 ±0.14) nmol/g,1.00± 0.00,0.27 ± 0.01,(0.28 ± 0.13)％; B group (8.37 ± 1.57) mmol/L,(102.43 ± 9.32) μmol/L,(5.96±1.43) nu/mg,(1.39±0.25) nmol/g,0.51 ±0.05,0.17±0.01,(19.36±2.56)％; C group(6.04 ± 1.52) mmol/L,(78.58 ± 13.63) μmol/L,(13.52 ± 1.47) nu/mg,(1.97±0.19) nmol/g,0.79±0.04,0.22±0.01,(11.47 ±3.61)％ the group A,BandC.Conclusion The present study demonstrates that the BBR could protect the kidney from the peroxide damage by increasing the expression of the SIRT1 which could promote the expression of the gene of SOD.And so the BBR plays a benefical role to resist peroxide damage and apoptosis in cholestatickidney injury.
