造血干细胞%微小核糖核酸%肝衰竭,急性%肿瘤坏死因子类%小鼠,近交BALB/c
造血榦細胞%微小覈糖覈痠%肝衰竭,急性%腫瘤壞死因子類%小鼠,近交BALB/c
조혈간세포%미소핵당핵산%간쇠갈,급성%종류배사인자류%소서,근교BALB/c
Hematopoietic stem cells%MicroRNAs%Liver failure,acute%Tumor necrosis factors%Mice,inbred BALB/c
目的 探讨间充质干细胞(MSC)对急性肝功能衰竭(ALF)小鼠肝组织中miRNA-155和TNF表达的影响,以及与MSC疗效间的关系.方法 采用随机数字表法和随机数余数分组法将BALB/c清洁级小鼠96只分为4组,即健康对照组、急性肝功能衰竭组、MSC治疗组和MSC预防组各24只.ALF组予以D-GalN 900 mg/kg+脂多糖10 μg/kg腹腔注射建立模型;MSC治疗组在900 mg/kg D-GalN+ 10 μg/kg脂多糖腹腔注射后2h,予以尾静脉注射MSC 1×106;MSC预防组在900mg/kgD-GalN+10 μg/kg脂多糖腹腔注射前予以尾静脉注射MSC 1×106;健康对照组予以0.9％氯化钠溶液1 mL腹腔注射.给药7h后每组处死小鼠12只,检测小鼠血清肝功能,ELISA法检测TNF水平,实时定量PCR检测肝组织miRNA-155、TNF mRNA.余下每组12只小鼠观察24 h生存率.组间均数比较用单因素方差分析,相关性分析采用Pearson和Spearman相关分析.结果 D GalN/脂多糖诱导7h后,MSC预防和MSC治疗组小鼠ALT水平均较ALF组降低,差异均有统计学意义(q值分别为13.20和4.58,均P＜0.01);MSC预防组和MSC治疗组AST水平均较ALF组降低,差异均有统计学意义(q值分别为18.90和9.83,均P＜0.01).与ALF组相比,MSC预防组和MSC治疗组血清TNF水平均有所降低,差异均有统计学意义(q值分别为14.70和3.95,均P＜0.01).健康对照组、ALF组、MSC治疗组和MSC预防组小鼠肝组织TNF mRNA表达水平分别为1.0±0.3、19.2±2.2、6.6±1.7和4.3±0.9,组间比较差异有统计学意义(F=72.24,P＜0.01).与ALF组相比,MSC预防组和MSC治疗组小鼠肝组织TNF mRNA表达水平均有下调,差异均有统计学意义(q值分别为31.40和21.72,均P＜0.01).与ALF组相比,MSC预防组和MSC治疗组小鼠肝组织miRNA-155水平均有下调,差异均有统计学意义(q值分别为19.40和9.58,均P＜0.01).ALF组小鼠肝组织miRNA 155上调和TNFmRNA诱导呈正相关(r＝0.734,P=0.007).MSC预防组和MSC治疗组miRNA-155和TNF mRNA的部分逆转亦呈正相关(r值分别为0.687和0.590,P值分别为0.014和0.044).给药后24 h,健康对照组、ALF组、MSC治疗组和MSC预防组各死亡0/12、10/12、6/12和3/12只,组间比较差异有统计学意义(x2＝19.078,P＜0.01).结论 在MSC干预小鼠ALF发病过程中,可以部分逆转上调的肝组织miRNA-155和TNF,且存在协同性,提示MSC治疗ALF可能通过对肝组织miRNA-155和TNF的调控发生作用.
目的 探討間充質榦細胞(MSC)對急性肝功能衰竭(ALF)小鼠肝組織中miRNA-155和TNF錶達的影響,以及與MSC療效間的關繫.方法 採用隨機數字錶法和隨機數餘數分組法將BALB/c清潔級小鼠96隻分為4組,即健康對照組、急性肝功能衰竭組、MSC治療組和MSC預防組各24隻.ALF組予以D-GalN 900 mg/kg+脂多糖10 μg/kg腹腔註射建立模型;MSC治療組在900 mg/kg D-GalN+ 10 μg/kg脂多糖腹腔註射後2h,予以尾靜脈註射MSC 1×106;MSC預防組在900mg/kgD-GalN+10 μg/kg脂多糖腹腔註射前予以尾靜脈註射MSC 1×106;健康對照組予以0.9％氯化鈉溶液1 mL腹腔註射.給藥7h後每組處死小鼠12隻,檢測小鼠血清肝功能,ELISA法檢測TNF水平,實時定量PCR檢測肝組織miRNA-155、TNF mRNA.餘下每組12隻小鼠觀察24 h生存率.組間均數比較用單因素方差分析,相關性分析採用Pearson和Spearman相關分析.結果 D GalN/脂多糖誘導7h後,MSC預防和MSC治療組小鼠ALT水平均較ALF組降低,差異均有統計學意義(q值分彆為13.20和4.58,均P＜0.01);MSC預防組和MSC治療組AST水平均較ALF組降低,差異均有統計學意義(q值分彆為18.90和9.83,均P＜0.01).與ALF組相比,MSC預防組和MSC治療組血清TNF水平均有所降低,差異均有統計學意義(q值分彆為14.70和3.95,均P＜0.01).健康對照組、ALF組、MSC治療組和MSC預防組小鼠肝組織TNF mRNA錶達水平分彆為1.0±0.3、19.2±2.2、6.6±1.7和4.3±0.9,組間比較差異有統計學意義(F=72.24,P＜0.01).與ALF組相比,MSC預防組和MSC治療組小鼠肝組織TNF mRNA錶達水平均有下調,差異均有統計學意義(q值分彆為31.40和21.72,均P＜0.01).與ALF組相比,MSC預防組和MSC治療組小鼠肝組織miRNA-155水平均有下調,差異均有統計學意義(q值分彆為19.40和9.58,均P＜0.01).ALF組小鼠肝組織miRNA 155上調和TNFmRNA誘導呈正相關(r＝0.734,P=0.007).MSC預防組和MSC治療組miRNA-155和TNF mRNA的部分逆轉亦呈正相關(r值分彆為0.687和0.590,P值分彆為0.014和0.044).給藥後24 h,健康對照組、ALF組、MSC治療組和MSC預防組各死亡0/12、10/12、6/12和3/12隻,組間比較差異有統計學意義(x2＝19.078,P＜0.01).結論 在MSC榦預小鼠ALF髮病過程中,可以部分逆轉上調的肝組織miRNA-155和TNF,且存在協同性,提示MSC治療ALF可能通過對肝組織miRNA-155和TNF的調控髮生作用.
목적 탐토간충질간세포(MSC)대급성간공능쇠갈(ALF)소서간조직중miRNA-155화TNF표체적영향,이급여MSC료효간적관계.방법 채용수궤수자표법화수궤수여수분조법장BALB/c청길급소서96지분위4조,즉건강대조조、급성간공능쇠갈조、MSC치료조화MSC예방조각24지.ALF조여이D-GalN 900 mg/kg+지다당10 μg/kg복강주사건립모형;MSC치료조재900 mg/kg D-GalN+ 10 μg/kg지다당복강주사후2h,여이미정맥주사MSC 1×106;MSC예방조재900mg/kgD-GalN+10 μg/kg지다당복강주사전여이미정맥주사MSC 1×106;건강대조조여이0.9％록화납용액1 mL복강주사.급약7h후매조처사소서12지,검측소서혈청간공능,ELISA법검측TNF수평,실시정량PCR검측간조직miRNA-155、TNF mRNA.여하매조12지소서관찰24 h생존솔.조간균수비교용단인소방차분석,상관성분석채용Pearson화Spearman상관분석.결과 D GalN/지다당유도7h후,MSC예방화MSC치료조소서ALT수평균교ALF조강저,차이균유통계학의의(q치분별위13.20화4.58,균P＜0.01);MSC예방조화MSC치료조AST수평균교ALF조강저,차이균유통계학의의(q치분별위18.90화9.83,균P＜0.01).여ALF조상비,MSC예방조화MSC치료조혈청TNF수평균유소강저,차이균유통계학의의(q치분별위14.70화3.95,균P＜0.01).건강대조조、ALF조、MSC치료조화MSC예방조소서간조직TNF mRNA표체수평분별위1.0±0.3、19.2±2.2、6.6±1.7화4.3±0.9,조간비교차이유통계학의의(F=72.24,P＜0.01).여ALF조상비,MSC예방조화MSC치료조소서간조직TNF mRNA표체수평균유하조,차이균유통계학의의(q치분별위31.40화21.72,균P＜0.01).여ALF조상비,MSC예방조화MSC치료조소서간조직miRNA-155수평균유하조,차이균유통계학의의(q치분별위19.40화9.58,균P＜0.01).ALF조소서간조직miRNA 155상조화TNFmRNA유도정정상관(r＝0.734,P=0.007).MSC예방조화MSC치료조miRNA-155화TNF mRNA적부분역전역정정상관(r치분별위0.687화0.590,P치분별위0.014화0.044).급약후24 h,건강대조조、ALF조、MSC치료조화MSC예방조각사망0/12、10/12、6/12화3/12지,조간비교차이유통계학의의(x2＝19.078,P＜0.01).결론 재MSC간예소서ALF발병과정중,가이부분역전상조적간조직miRNA-155화TNF,차존재협동성,제시MSC치료ALF가능통과대간조직miRNA-155화TNF적조공발생작용.
Objective To explore the effects of the mesenchymal stem cells (MSC) on the microRNA 155 (miRNA-155) and tumor necrosis factor (TNF) expressions in liver tissue of mice with acute liver failure (ALF),and to explore the relationship between miRNA 155/TNF and the efficacy of MSC.Methods Using a random number table method,and the remainder grouping method of random numbers.A total of 96 BALB/c mice of clean grade were randomly divided into four groups,namely groups of healthy control (intraperitoneal injection of 0.9 ％ NaCl,1 mI),ALF (intraperitoneal injection of D-galactosamine (D-GalN),900 mg/kg body weight,and lipopolysaccharides,10 mg/kg body weight),MSC treatment (1 × 106 MSC tail intravenously injected 2 h after intraperitoneal injection of D-GalN and lipopolysaccharides) and MSC pretreatment (1)× 106 MSC tail intravenously injected before intraperitoneal injection of D-GalN and lipopolysaccharides).Twelve of the 24 mice in each group were sacrificed 7 h after intraperitoneal administration.Liver function,serum TNF level,miRNA-155 and TNF mRNA of liver tissue were detected subsequently.Survival rate at 24 h was observed in the remaining 12 mice in each group.Mean comparison between groups was conducted with ANOVA,and correlation analysis was performed using Pearson and Spearman correlation analysis.Results Seven hours after D-GalN/lipopolysaccharides induction,alanine aminotransferase levels of MSC treatment and MSC pretreatment were significantly lower when compared with those of ALF (q=13.20 and 4.58,respectively; both P＜0.01) ; aspartate aminotransferase levels decreased in MSC treatment and MSC pretreatment groups compared to ALF group,with significant statistical differences (q=18.90 and 9.83,respectively; both P＜0.01).Compared with ALF,serum levels of TNF decreased in MSC treatment and MSC pretreatment and the difference was statistically significant (q=14.70 and 3.95,respectively; both P＜0.01).The TNF mRNA expressions in liver tissue of the healthy control,ALF,MSC treatment and MSC pretreatment were 1.0±0.3,19.2±2.2,6.6±1.7 and 4.3±0.9,respectively.The difference between groups was statistically significant (F 72.24,P＜ 0.01).The TNF mRNA expressions of MSC treatment and MSC pretreatment were down-regulated in comparison with ALF,which showed statistical difference (q=31.40 and 21.72,respectively; both P＜0.01).Compared with ALF,miRNA 155 in liver tissue of MSC treatment and MSC pretreatment were lowered and the differences were significant (q=19.40 and 9.58,respectively; both P＜0.01).The positive correlation between miRNA 155 and TNF mRNA in liver tissue was confirmed in ALF (r＝0.734,P=0.007),MSC treatment (r=0.590,P=0.044) and MSC pretreatment (r=0.687,P=0.014).24 h after administration,the mortalities of the healthy control,ALF,MSC treatment and MSC pretreatment were 0/12,10/12,6/12 and 3/12,respectively.The difference between groups was statistically significant (x2 =19.078,P ＜ 0.01).Conclusion With MSC intervention in ALF,up regulated miRNA-155 and TNF expressions in liver tissue could be synergetic and partially reversed by MSC,suggesting that MSC treatment for ALF may play an important role via regulating miRNA-155 and TNF.
