中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
12期
2690-2692
,共3页
李静静%司晓芸%刘曦明%吴小燕
李靜靜%司曉蕓%劉晞明%吳小燕
리정정%사효예%류희명%오소연
肾缺血%再灌注损伤%骨髓间充质干细胞%CXC趋化因子受体-4
腎缺血%再灌註損傷%骨髓間充質榦細胞%CXC趨化因子受體-4
신결혈%재관주손상%골수간충질간세포%CXC추화인자수체-4
Renal ischemia%Reperfusion injury%Bone mesenchymal stem cells%CXC chemokine receptor 4
目的 观察缺血再灌注(I/R)肾损伤微环境对大鼠骨髓间充质干细胞(BMSCs)表面CXC趋化因子受体-4(CXCR4)表达及肾保护性细胞因子分泌的影响.方法 制作不同浓度I/R肾损伤组织匀浆,与BMSCs共培养.激光共聚焦显微镜观察BMSCs表面CXCR4受体表达;Transwell实验检测BMSCs向基质细胞源性因子-1(SDF-1)的迁移能力;逆转录-聚合酶链反应(RT-PCR)法检测基质细胞源性因子-1α(SDF-1α)、CXCR4、肝细胞生长因子(HGF) mRNA表达.结果 I/R肾损伤匀浆诱导BMSCs表面CXCR4表达明显上升(P＜0.01);该表达可被CXCR4中和抗体所抑制(P＜0.05).CXCR4表达增加后,BMSCs向SDF-1的迁移能力增强[(26.72±5.61)、(30.44 ±6.03)、(38.92 ±6.79)/cm2比(18.47 ±5.02)/cm2,P＜0.01].I/R肾损伤匀浆能够显著上调SDF-1α mRNA表达,但各组间表达差异无统计学意义(P＞0.05).肾损伤匀浆能够显著上调CXCR4mRNA(3.11 ±0.24比2.02 ±0.17,P＜0.01)和HGF mRNA(3.01 ±0.46比1.99 ±0.21,P＜0.01)表达,表达随匀浆浓度增加而升高,升高的表达可被CXCR4中和抗体所抑制(1.86±0.11,1.78±0.22,P＜0.05).结论 I/R肾损伤微环境能够通过提高BMSCs表面CXCR4受体表达促进BBMSCs迁移,活化的BMSCs上调保护性细胞因子表达,形成有利于肾损伤修复的微环境.
目的 觀察缺血再灌註(I/R)腎損傷微環境對大鼠骨髓間充質榦細胞(BMSCs)錶麵CXC趨化因子受體-4(CXCR4)錶達及腎保護性細胞因子分泌的影響.方法 製作不同濃度I/R腎損傷組織勻漿,與BMSCs共培養.激光共聚焦顯微鏡觀察BMSCs錶麵CXCR4受體錶達;Transwell實驗檢測BMSCs嚮基質細胞源性因子-1(SDF-1)的遷移能力;逆轉錄-聚閤酶鏈反應(RT-PCR)法檢測基質細胞源性因子-1α(SDF-1α)、CXCR4、肝細胞生長因子(HGF) mRNA錶達.結果 I/R腎損傷勻漿誘導BMSCs錶麵CXCR4錶達明顯上升(P＜0.01);該錶達可被CXCR4中和抗體所抑製(P＜0.05).CXCR4錶達增加後,BMSCs嚮SDF-1的遷移能力增彊[(26.72±5.61)、(30.44 ±6.03)、(38.92 ±6.79)/cm2比(18.47 ±5.02)/cm2,P＜0.01].I/R腎損傷勻漿能夠顯著上調SDF-1α mRNA錶達,但各組間錶達差異無統計學意義(P＞0.05).腎損傷勻漿能夠顯著上調CXCR4mRNA(3.11 ±0.24比2.02 ±0.17,P＜0.01)和HGF mRNA(3.01 ±0.46比1.99 ±0.21,P＜0.01)錶達,錶達隨勻漿濃度增加而升高,升高的錶達可被CXCR4中和抗體所抑製(1.86±0.11,1.78±0.22,P＜0.05).結論 I/R腎損傷微環境能夠通過提高BMSCs錶麵CXCR4受體錶達促進BBMSCs遷移,活化的BMSCs上調保護性細胞因子錶達,形成有利于腎損傷脩複的微環境.
목적 관찰결혈재관주(I/R)신손상미배경대대서골수간충질간세포(BMSCs)표면CXC추화인자수체-4(CXCR4)표체급신보호성세포인자분비적영향.방법 제작불동농도I/R신손상조직균장,여BMSCs공배양.격광공취초현미경관찰BMSCs표면CXCR4수체표체;Transwell실험검측BMSCs향기질세포원성인자-1(SDF-1)적천이능력;역전록-취합매련반응(RT-PCR)법검측기질세포원성인자-1α(SDF-1α)、CXCR4、간세포생장인자(HGF) mRNA표체.결과 I/R신손상균장유도BMSCs표면CXCR4표체명현상승(P＜0.01);해표체가피CXCR4중화항체소억제(P＜0.05).CXCR4표체증가후,BMSCs향SDF-1적천이능력증강[(26.72±5.61)、(30.44 ±6.03)、(38.92 ±6.79)/cm2비(18.47 ±5.02)/cm2,P＜0.01].I/R신손상균장능구현저상조SDF-1α mRNA표체,단각조간표체차이무통계학의의(P＞0.05).신손상균장능구현저상조CXCR4mRNA(3.11 ±0.24비2.02 ±0.17,P＜0.01)화HGF mRNA(3.01 ±0.46비1.99 ±0.21,P＜0.01)표체,표체수균장농도증가이승고,승고적표체가피CXCR4중화항체소억제(1.86±0.11,1.78±0.22,P＜0.05).결론 I/R신손상미배경능구통과제고BMSCs표면CXCR4수체표체촉진BBMSCs천이,활화적BMSCs상조보호성세포인자표체,형성유리우신손상수복적미배경.
Objective To explore the effect of ischemia reperfusion-injured (IRI) kidney homogenate in vitro on CXC chemokine receptor 4 (CXCR4) expression in bone mesenchymal stem cells (BMSCs) and renal protective growth factors paracrine.Methods The renal homogenate supernatant of IRI model was built and co-cultured with BMSCs.CXCR4 protein expression was detected by laser focal microscope,chemotactic ability of BMSCs to stromal cells derived factor-1α (SDF-1α) was investigated by Transwell test,and the mRNA expression of SDF-1α,CXCR4 and hepatocyte growth factor (HGF) was detected by reverse transcription-polymerase chain reaction (RT-PCR).Results CXCR4 protein expression was increased significantly after co-culture of BMSCs with IRI homogenate [2.54 ± 0.22 vs.1.26 ± 0.18,P ＜0.01),which ould be decreased by CXCR4 neutralizing antibody,and the chemotactic ability of BMSCs to SDF-1 increased at the same time [(26.72 ±5.61),(30.44 ±6.03),(38.92 ±6.79)/cm2 vs.(18.47 ±5.02)/cm2,P ＜0.01].SDF-1α mRNA expression was increased in IRI group,but had no significan difference among groups (P ＞ 0.05).CXCR4 mRNA (3.11 ± 0.24 vs.2.02 ± 0.17,P ＜ 0.01) and HGF mRNA (3.01 ±0.46 vs.1.99 ±0.21,P ＜0.01) expression was increased significantly with the homogenate,and the expression could be restrained by CXCR4 neutrlizing antibody (1.86 ± 0.11 and 1.78 ± 0.22,P ＜0.05).Conclusion IRI kidney microenvironment can promote chemotactic ability of BMSCs by increasing BMSCs surface CXCR4 expression,and then CXCR4 + BMSCs possessed the paracrine capabilities which promoted the recovery of the injured kidney.
