目的 探讨骨髓间充质干细胞(MSC)移植和动员对重症急性胰腺炎(SAP)大鼠急性肾损伤的保护作用.方法 240只SD大鼠按照随机数字表法分为假手术组、模型组、干细胞移植组、干细胞动员组和联合组,每组48只.腹腔注射L-精氨酸制作SAP大鼠模型.假手术组在制作SAP大鼠模型后腹腔注射生理盐水,干细胞移植组制作SAP大鼠模型后6h经股静脉注入自体MSC 1.2 ml,干细胞动员组制作SAP大鼠模型前连续3d皮下注入重组人粒细胞集落刺激因子(G-CSF) 40 μg/kg,联合组则联合应用MSC和G-CSF.各组大鼠再按术后不同时相点分为12、24、48、72 h亚组,每组12只.在术后相应时相点观察各组大鼠的存活情况,肾脏组织的病理变化,肾小管上皮细胞Bax和Bcl-2蛋白的表达情况和细胞凋亡指数,检测血清中TNF-α、IL-6、血清尿素氮(BUN)、肌酐(Cr)、LDH、C反应蛋白(CRP)的含量.采用单因素方差分析各组间的指标,两两比较用SNK-q检验,大鼠存活情况用Fisher确切概率法.结果 假手术组大鼠全部存活.模型组大鼠术后48、72 h分别存活11只和8只.干细胞移植组、干细胞动员组和联合组术后48 h前未见大鼠死亡,术后72 h分别存活11、10和11只,与模型组比较,差异无统计学意义(P＞0.05).各治疗组术后肾脏组织病理变化均较模型组减轻,联合组损伤减轻最为明显.术后12 ～72 h肾小管上皮细胞Bax蛋白、Bcl-2蛋白、肾小管细胞凋亡指数变化情况:模型组分别为12.80±1.78 ～20.30±2.40、4.34±1.20 ～3.03±1.06、12.65％±2.31％ ～35.10％±5.54％;干细胞移植组分别为9.68±2.11～17.01±2.54、5.57±1.35～4.13±1.05、6.20％±1.53％ ～ 17.50％±2.80％;干细胞动员组分别为10.05±2.17～16.81±2.55、5.49±1.48～4.19±1.05、6.41％±1.64％ ～ 17.14％±2.27％;联合组分别为8.33±2.06～14.03±2.27、6.60±2.11 ～5.63±1.52、5.80％±1.52％～12.30％±2.43％.联合组术后24、72 h Bax蛋白,48、72 h Bc1-2蛋白,24、48、72 h凋亡指数与干细胞移植组和干细胞动员组比较,差异有统计学意义(P＜0.05);干细胞移植组与干细胞动员组比较,差异无统计学意义(P＞0.05);各治疗组术后12 ～72 h炎症因子及肾功能指标较模型组不同程度降低,以联合组最明显.联合组术后72 hTNF-α含量,48、72 h IL-6含量,48、72 h BUN含量,48、72 h Cr含量,24、48、72 h LDH含量,72 h CRP含量与干细胞移植组和干细胞动员组比较,差异有统计学意义(P＜0.05);干细胞移植组与于细胞动员组比较,差异无统计学意义(P＞0.05).结论 自体MSC移植与动员能有效减轻SPA大鼠的肾损伤,可能与MSC参与组织的病理再生修复、抗炎症及抑制细胞凋亡等机制有关.
目的 探討骨髓間充質榦細胞(MSC)移植和動員對重癥急性胰腺炎(SAP)大鼠急性腎損傷的保護作用.方法 240隻SD大鼠按照隨機數字錶法分為假手術組、模型組、榦細胞移植組、榦細胞動員組和聯閤組,每組48隻.腹腔註射L-精氨痠製作SAP大鼠模型.假手術組在製作SAP大鼠模型後腹腔註射生理鹽水,榦細胞移植組製作SAP大鼠模型後6h經股靜脈註入自體MSC 1.2 ml,榦細胞動員組製作SAP大鼠模型前連續3d皮下註入重組人粒細胞集落刺激因子(G-CSF) 40 μg/kg,聯閤組則聯閤應用MSC和G-CSF.各組大鼠再按術後不同時相點分為12、24、48、72 h亞組,每組12隻.在術後相應時相點觀察各組大鼠的存活情況,腎髒組織的病理變化,腎小管上皮細胞Bax和Bcl-2蛋白的錶達情況和細胞凋亡指數,檢測血清中TNF-α、IL-6、血清尿素氮(BUN)、肌酐(Cr)、LDH、C反應蛋白(CRP)的含量.採用單因素方差分析各組間的指標,兩兩比較用SNK-q檢驗,大鼠存活情況用Fisher確切概率法.結果 假手術組大鼠全部存活.模型組大鼠術後48、72 h分彆存活11隻和8隻.榦細胞移植組、榦細胞動員組和聯閤組術後48 h前未見大鼠死亡,術後72 h分彆存活11、10和11隻,與模型組比較,差異無統計學意義(P＞0.05).各治療組術後腎髒組織病理變化均較模型組減輕,聯閤組損傷減輕最為明顯.術後12 ～72 h腎小管上皮細胞Bax蛋白、Bcl-2蛋白、腎小管細胞凋亡指數變化情況:模型組分彆為12.80±1.78 ～20.30±2.40、4.34±1.20 ～3.03±1.06、12.65％±2.31％ ～35.10％±5.54％;榦細胞移植組分彆為9.68±2.11～17.01±2.54、5.57±1.35～4.13±1.05、6.20％±1.53％ ～ 17.50％±2.80％;榦細胞動員組分彆為10.05±2.17～16.81±2.55、5.49±1.48～4.19±1.05、6.41％±1.64％ ～ 17.14％±2.27％;聯閤組分彆為8.33±2.06～14.03±2.27、6.60±2.11 ～5.63±1.52、5.80％±1.52％～12.30％±2.43％.聯閤組術後24、72 h Bax蛋白,48、72 h Bc1-2蛋白,24、48、72 h凋亡指數與榦細胞移植組和榦細胞動員組比較,差異有統計學意義(P＜0.05);榦細胞移植組與榦細胞動員組比較,差異無統計學意義(P＞0.05);各治療組術後12 ～72 h炎癥因子及腎功能指標較模型組不同程度降低,以聯閤組最明顯.聯閤組術後72 hTNF-α含量,48、72 h IL-6含量,48、72 h BUN含量,48、72 h Cr含量,24、48、72 h LDH含量,72 h CRP含量與榦細胞移植組和榦細胞動員組比較,差異有統計學意義(P＜0.05);榦細胞移植組與于細胞動員組比較,差異無統計學意義(P＞0.05).結論 自體MSC移植與動員能有效減輕SPA大鼠的腎損傷,可能與MSC參與組織的病理再生脩複、抗炎癥及抑製細胞凋亡等機製有關.
목적 탐토골수간충질간세포(MSC)이식화동원대중증급성이선염(SAP)대서급성신손상적보호작용.방법 240지SD대서안조수궤수자표법분위가수술조、모형조、간세포이식조、간세포동원조화연합조,매조48지.복강주사L-정안산제작SAP대서모형.가수술조재제작SAP대서모형후복강주사생리염수,간세포이식조제작SAP대서모형후6h경고정맥주입자체MSC 1.2 ml,간세포동원조제작SAP대서모형전련속3d피하주입중조인립세포집락자격인자(G-CSF) 40 μg/kg,연합조칙연합응용MSC화G-CSF.각조대서재안술후불동시상점분위12、24、48、72 h아조,매조12지.재술후상응시상점관찰각조대서적존활정황,신장조직적병리변화,신소관상피세포Bax화Bcl-2단백적표체정황화세포조망지수,검측혈청중TNF-α、IL-6、혈청뇨소담(BUN)、기항(Cr)、LDH、C반응단백(CRP)적함량.채용단인소방차분석각조간적지표,량량비교용SNK-q검험,대서존활정황용Fisher학절개솔법.결과 가수술조대서전부존활.모형조대서술후48、72 h분별존활11지화8지.간세포이식조、간세포동원조화연합조술후48 h전미견대서사망,술후72 h분별존활11、10화11지,여모형조비교,차이무통계학의의(P＞0.05).각치료조술후신장조직병리변화균교모형조감경,연합조손상감경최위명현.술후12 ～72 h신소관상피세포Bax단백、Bcl-2단백、신소관세포조망지수변화정황:모형조분별위12.80±1.78 ～20.30±2.40、4.34±1.20 ～3.03±1.06、12.65％±2.31％ ～35.10％±5.54％;간세포이식조분별위9.68±2.11～17.01±2.54、5.57±1.35～4.13±1.05、6.20％±1.53％ ～ 17.50％±2.80％;간세포동원조분별위10.05±2.17～16.81±2.55、5.49±1.48～4.19±1.05、6.41％±1.64％ ～ 17.14％±2.27％;연합조분별위8.33±2.06～14.03±2.27、6.60±2.11 ～5.63±1.52、5.80％±1.52％～12.30％±2.43％.연합조술후24、72 h Bax단백,48、72 h Bc1-2단백,24、48、72 h조망지수여간세포이식조화간세포동원조비교,차이유통계학의의(P＜0.05);간세포이식조여간세포동원조비교,차이무통계학의의(P＞0.05);각치료조술후12 ～72 h염증인자급신공능지표교모형조불동정도강저,이연합조최명현.연합조술후72 hTNF-α함량,48、72 h IL-6함량,48、72 h BUN함량,48、72 h Cr함량,24、48、72 h LDH함량,72 h CRP함량여간세포이식조화간세포동원조비교,차이유통계학의의(P＜0.05);간세포이식조여우세포동원조비교,차이무통계학의의(P＞0.05).결론 자체MSC이식여동원능유효감경SPA대서적신손상,가능여MSC삼여조직적병리재생수복、항염증급억제세포조망등궤제유관.
Objective To investigate the protective effects of bone marrow mesenchymal stem cells transplantation (MSCT) and mobilization on severe acute pancreatitis (SAP) with acute renal injury.Methods A total of 240 SD rats were randomly divided into sham operation group ( n =48 ),model control group ( n =48 ),MSCT group ( n =48),bone marrow mesenchymal stem cells mobilization (MSCM) group ( n =48) and MSCT +MSCM group ( n =48 ) according to the random number table.Rat models of SAP were made by peritoneal injection of L-arginine.Rats in the MSCT group were injected with 1.2 ml of bone marrow mesenchymai stem cells via femoral vein at 6 hours after SAP model establishment; rats in the MSCM group were subcutaneously injected with 40 μg/kg of granulocyte-colony stimulating factor (G-CSF) at 3 days before SAP model establishment; rats in the MSCT + MSCM group were injected with 1.2 ml of MSC and 40 μg/kg of G-CSF simultaneously; rats in the sham operation group were injected with equal volume of normal saline.According to different time points after operation,rats in each group were subdivided into 12 h,24 h,48 h and 72 h groups (n =12).At each time points after operation,the mortality rate,pathological changes of renal tissue,expression of Bax protein,Bcl-2 protein and apoptosis indexes of renal tubular epithelium cells were observed.The contents of tumor necrotic factor-α (TNF-α),interleukin-6 (IL-6),blood urea nitrogen (BUN),creatinine (Cr),lactate dehydrogenase (LDH) and C-reactive protein (CRP) were determined.All data were analyzed by using SNK-q test,Fisher exact probability and analysis of variance.Results All rats in the sham operation group were survived.The numbers of rats in the model control group survived at postoperative 48 hours and 72 hours were 11 and 8,respectively.No rat died at postoperative 48 hours in the MSCT group,MSCM group and MSCT + MSCM group.The numbers of rats survived at postoperative 72 hours in the MSCT group,MSCM group and MSCT + MSCM group were 11,10 and 11,which were not significantly different from the number of survived rats in the model control group (P ＞0.05).The pathological injuries of renal tissues were relieved in the MSCT group,MSCM group and MSCT + MSCM group when compared with model control group.The expression of Bax protein,Bc1-2 protein,renal tubular epithelium cell apoptosis indexes at 12-72 hours were 12.80 + 1.78-20.30 + 2.40,4.34 + 1.20-3.03 ± 1.06,12.65％ ±2.31％-35.10％ ± 5.54％ in the model control group,9.68 ± 2.11-17.01 ± 2.54,5.57 ± 1.35-4.13 + 1.05,6.20％ ± 1.53％- 17.50％ ± 2.80％ in the MSCT group,10.05 ± 2.17-16.81 ± 2.55,5.49 ± 1.48-4.19 ±1.05,6.41％± 1.64％-17.14％±2.27％ in the MSCM group,8.33 ±2.06-14.03 ±2.27,6.60 ±2.11-5.63 ±1.52,5.80％ ± 1.52％-12.30％ ±2.43％ in the MSCT + MSCT group.There were significant differences in the expressions of Bax protein at 24 and 72 hours,Bcl-2 protein at 48 and 72 hours,renal tubular epithelium cell apoptosis index at 24,48 and 72 hours between the MSCT group,MSCM group and MSCT + MSCM group ( P ＜0.05 ),but no significant difference was found between the MSCT group and the MSCM group ( P ＞ 0.05 ).The contents of TNF-α,IL-6,BUN,Cr,LDH,CRP in the MSCT group,MSCM group and MSCT + MSCM group were decreased when compared with those in the model control group,and a significant decrease of the 6 factors was observed in the MSCT + MSCM group.There were significant difference in the content of TNF-α at 72 hours,IL-6,BUN and Cr at 48 and 72 hours,LDH at 24,48 and 72 hours and CRP at 72 hours between the MSCT group,MSCM group and MSCT + MSCM group (P ＜0.05),while no significant difference was observed between the MSCT group and the MSCM group (P ＞ 0.05).Conclusion MSCT and MSCM can significantly protect acute renal injury in the progress of SAP,the probable mechanisms are pathological regeneration,anti-inflammatory effect and apoptosis inhibition of mesenchymal stem cells.
