尼克酰胺对人脐带间充质干细胞的保护效应

Protective effect of nicotinic acid amide on human umbilical cord mesenchymal stem cells

目的:探讨尼克酰胺(nicotinic acid amide,NAA)减轻人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,h UC-MSCs)输注损伤的效应。方法:空白对照组为0.3 m L生理盐水、MSC组为0.3 m L 1×106CFSE标记的h UC-MSCs,MSC+NAA组为0.3 m L 10 mmol/L NAA处理24 h的1×106CFSE标记的h UC-MSCs,分别与2.7 m L正常人全血混匀,注入改良Chandler Loop内,在37℃下,20 m L/min循环1 h,检测循环前后血小板、白细胞、h UC-MSCs和C3a的变化。结果:血小板消耗率空白对照组为(29.96±10.88)%、MSC组为(77.76±19.29)%、MSC+NAA组为(50.13±18.10)%;白细胞消耗率空白对照组为(37.82±13.81)%、MSC组为(64.57±17.08)%、MSC+NAA组为(41.52±17.26)%,MSC组和MSC+NAA组与空白对照组比较差异均有统计学意义(P<0.05)。MSC+NAA组的h UC-MSCs存活率较MSC组高(P<0.05)。空白对照、MSC和MSC+NAA组的C3a含量分别为(206.27±58.10)μg/L、(230.47±39.61)μg/L和(208.37±40.66)μg/L。结论:h UC-MSCs与正常人全血在改良Chandler Loop内共循环1 h可模拟血液介导即刻炎症反应(instant blood-mediated inflammatory reaction,IBMIR),大量损耗MSCs和血液成分,导致C3a上升。NAA可抑制IBMIR、降低血液成分的损耗、提高h UC-MSCs的存活率,提示NAA能减轻MSCs的输注损伤。

AIM： To investigate the effect of nicotinic acid amide （NAA） on the infusion damage of human umbilical cord mesenchymal stem cells （hUC-MSCs） under the condition of instant blood-mediated inflammatory reaction （IBMIR）. METHODS： Normal peripheral blood without anticoagulant at volume of 2.7 mL was mixed with 0. 3 mL phys- iological saline （as blank group） , CFSE labeled hUC-MSCs （1 × 10^6 cells in 0. 3 mL as MSC group） and CFSE labeled hUC-MSCs （1 ×10^6 cells in 0. 3 mL） preprocessed with NAA at concentration of 10 mmol/L for 24 h （as MSC ＋ NAA group） , respectively. The mixture was immediately injected into the improved Chandler Loop model, placed in 37 % water bath, and then started the peristaltic pump at the speed of 20 mL/min for 1 h. The number of CFSE labeled hUC-MSCs, platelets, white blood ceils were counted and the concentration of complement C3a was measured before and after cycling, respectively. RESULTS： After 1 h circulation, the platelet dissipation rate were （29.96 ± 10. 88 ）% in blank group, （77.76 ± 19. 29）% in MSC group all and （50. 13 ± 18.10）% in MSC ＋ NAA group; and the leukocyte counts were （37.82 ± 13.81 ）% in blank group, （64. 57 ± 17. 08 ）% in MSC group and （41.52 ± 17.26）% in MSC ＋ NAA group. Compared with blank group, the differences of the dissipation rates in MSC group and MSC ＋ NAA group all had statistical significance. The hUC-MSCs relative survival rate in MSC ＋ NAA group was higher than that in MSC group. C3a concentra- tions in blank group, MSC group and MSC ＋ NAA group were （ 206. 27 ± 58. 10）, （ 230. 47± 39. 61 ） and （ 208.37 ± 40. 66） μg/L, respectively. CONCLUSION： Co-circulating the mixture of hUC-MSCs with normal peripheral blood with- out anticoagulant in the improved Chandler Loop for 1 h depletes a large number of hUC-MSCs and blood components, and increases C3a, suggesting that this model can induce IBMIR. NAA has a protective effect on the hUC-MSCs in the infusion damage by inhibiting IBMIR, reducing the wastage of the blood components and enhancing the survival rate of the hUC- MSCs.