渗透压对人体关节软骨损伤后软骨细胞修复的机制  

Mechanism of Osmotic Pressure on Repairing Chondrocytes after Injury of Human Articular Cartilage

作　　者：谭显春[1] 李欣 赖超超[1] 符纯锋 吴晓均[1] 雷孝勇 王海珠[1] Tan Xianchun;Li Xin;Lai Chaochao;Fu Chunfeng;Wu Xiaojun;Lei Xiaoyong;Wang Haizhu(Dazhou Integrated TCM and Western Medicine Hospital,Dazhou,635000)

机构地区：[1]达州市中西医结合医院

出　　处：《基因组学与应用生物学》2019年第10期4826-4831,共6页Genomics and Applied Biology

摘　　要：为了探究灌注溶液渗透压对机械损伤后关节软骨细胞存活率以及对代谢状态的影响,本研究以全膝关节置换治疗骨关节炎的患者中采集骨软骨外植体,切开其全层厚度,建立机械损伤模型。将软骨外植体在具有不同渗透压的灌溉溶液(盐水和平衡盐)中孵育(A:185 mOsm/L;B:285 mOsm/L;C:385 mOsm/L;D:585 mOsm/L)2.5 h。用激光共聚焦显微镜定量细胞死亡率(100%×死细胞数/死细胞和活细胞数)。以末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)测定受损软骨的凋亡指数。用分光光度计在530 nm处测定蛋白多糖洗脱量,并利用实时PCR定量HIF-1α和Ⅱ型胶原mRNA的产量。原位死亡软骨细胞主要位于机械损伤后受损软骨边缘的浅表切向区域。随着渗透压的升高,细胞死亡率下降,蛋白多糖洗脱量逐渐减少。不同渗透压下TUNEL试验的凋亡指数差异无统计学意义(p>0.05)。与生理盐水组相比,平衡盐组的细胞死亡率显著下降(p<0.01),蛋白多糖洗脱量也显著下降(p<0.05)。冲洗溶液的渗透压对机械损伤后软骨细胞的存活率和代谢状态有重要调节作用,软骨细胞死亡并不是由细胞凋亡导致。增加灌洗液的渗透压可增加对软骨的保护性,可减少软骨细胞的死亡,同时能减少对代谢抑制和蛋白多糖的洗脱量,最终防止软骨退化和促进整合修复。To investigate the effect of perfusion solution osmotic pressure on the survival rate of articular chondrocytes and the metabolic state after mechanical injury.Osteochondral explants were harvested from patients with osteoarthritis treated with total knee arthroplasty.The full thickness of the cartilage was cut and a mechanical injury model was established.The cartilage explants were incubated(A:185 mOsm/L;B:285 mOsm/L;C:385 mOsm/L;D:585 mOsm/L)in an irrigation solution(saline and balanced salt)with different osmotic pressures for 2.5 hours.Cell death was quantified by laser confocal microscopy(100%×dead cells/dead cells and viable cells).The apoptotic index of damaged cartilage was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL).The proteoglycan elution amount was measured at 530 nm with a spectrophotometer,and the yield of HIF-1αand typeⅡcollagen mRNA was quantified by real-time PCR.The in situ dead chondrocytes were mainly located in the superficial tangential region of the damaged cartilage edge after mechanical injury.As the osmotic pressure increases,the cell death rate decreases and the amount of proteoglycan eluted gradually decreases.There was no significant difference in the apoptotic index of TUNEL test under different osmotic pressures(p>0.05).Compared with the saline group,the cell death rate of the balanced salt group was significantly decreased(p<0.01),and the proteoglycan elution amount was also significantly decreased(p<0.05).The osmotic pressure of the washing solution plays an important role in regulating the survival rate and metabolic state of chondrocytes after mechanical injury.Chondrocyte death is not caused by apoptosis.Increasing the osmotic pressure of the lavage fluid can increase the protection of the cartilage,reduce the death of chondrocytes,reduce the amount of metabolism inhibition and proteoglycan elution,and ultimately prevent cartilage degradation and promote integrated repair.

关 键 词：渗透压 TUNEL 软骨细胞 关节软骨 HIF-1Α 

分 类 号：R73[医药卫生—肿瘤]