人脐带华通胶间充质干细胞对白细胞介素1β诱导髓核细胞退变的保护作用

Human umbilical cord Wharton's jelly-derived mesenchymal stem cells protect against interleukin-1beta induced degeneration of nucleus pulposus cells

背景:人脐带华通胶间充质干细胞移植到犬退变椎间盘模型中可存活,恢复部分椎间盘高度,但其对炎性损伤后髓核细胞是否有修复治疗作用目前还没有文献报道。目的:观察人脐带华通胶间充质干细胞对白细胞介素1β诱导大鼠髓核细胞退变的保护作用。方法:将第3代SD大鼠髓核细胞分3组培养,对照组用含胎牛血清的培养基培养24 h,然后换完全培养基培养24 h;白细胞介素1β组以含白细胞介素1β和胎牛血清的培养基培养24 h,然后换完全培养基培养24 h;实验组先以含白细胞介素1β和胎牛血清的培养基培养24 h,再更换完全培养基与人脐带华通胶间充质干细胞非直接共培养24 h。采用Annexin V/PI染色后流式细胞仪测定3组髓核细胞凋亡率,RT-PCR测定各组髓核细胞基质金属蛋白酶4、基质金属蛋白酶3、基质金属蛋白酶抑制剂1的表达,caspase试剂盒检测caspase-3、caspase-8和caspase-9活性。结果与结论:(1)白细胞介素1β组、实验组细胞凋亡率高于对照组(P<0.05),实验组低于白细胞介素1β组(P<0.05);(2)白细胞介素1β组、实验组基质金属蛋白酶4、基质金属蛋白酶3表达量高于对照组(P<0.05),基质金属蛋白酶抑制剂1表达量低于对照组(P<0.05);实验组基质金属蛋白酶4、基质金属蛋白酶3表达量低于白细胞介素1β组,基质金属蛋白酶抑制剂1表达量高于白细胞介素1β组(P<0.05);(3)白细胞介素1β组、实验组caspase-3、caspase-8、caspase-9相对活性高于对照组(P<0.05),实验组caspase-3、caspase-8、caspase-9相对活性低于白细胞介素1β组(P<0.05);(4)结果表明,人脐带华通胶间充质干细胞对白细胞介素1β诱导的髓核细胞退变凋亡有明显的抑制作用。

BACKGROUND:Human umbilical cord Wharton’s jelly-derived mesenchymal stem cells(hWJCs)via transplantation are able to survive in a dog model of intervertebral disc degeneration and to partially restore the height of the intervertebral disc.However,there is no study on whether hWJCs can repair nucleus pulposus cells after inflammatory injury as yet.OBJECTIVE:To observe the protective effect of hWJCs on interleukin-1βinduced rat nucleus pulposus degeneration.METHODS:Nucleus pulposus cells at passage3from Sprague-Dawley rats were divided into three groups:(1)normal control group cultured in culture medium containing fetal bovine serum for24hours,and then cultured in complete medium for24hours;(2)intervention group with interleukin-1βintervention for24hours,followed by culture in the complete medium for24hours;(3)co-culture group with interleukin-1βintervention for24hours,followed by culture inthe complete medium for non-direct co-culture with hWJCs for24hours.Annexin V-FITC/propidium iodide(PI)double staining and flow cytometry were used in succession to measure the apoptosis rate of nucleus pulposus cells in each group.The ADAMTS-4,MMPs-3and TIMP-1expression in nucleus pulposus cells was observed though RT-PCR.The activities of caspase-3,caspase-8and caspase-9were determined by caspase kit.