Wang Zheyan, Wang Wenya, Zhang Liu, Tian Faming, Cheng Tan, Yang Lin
Department of Pathology, North China Coal Medical College, Tangshan Hebei, 063000, P.R. China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2011 Jan;25(1):50-5.
To investigate the feasibility of alendronate (ALN) in treating osteoarthritis (OA) by observing the effects of ALN on interleukin 1beta (IL-1beta) induced chondrocytes of rat in vitro.
The chondrocytes of knee articular surface from 15 SD rats (1-month-old, female or male, weighing 100-150 g) were cultured. The chondrocytes were observed by inverted phase contrast microscope and identified by toluidine blue staining and HE staining. The third passage chondrocytes were divided into 3 groups: the chondrocytes were cultured with DMEM for 5 days in group A, with 10 ng/mL IL-1beta for 2 days and with DMEM for 3 days in group B, and with 10 ng/mL IL-1beta for 2 days and with 1 x 10(6) mol/L ALN for 3 days in group C. Immunocytochemistry and real-time PCR were performed to determine the expression levels of collagen type II (Col II), matrix metalloproteinase 13 (MMP-13), and p-catenin.
Toluidine blue staining proved that the cultured cells were chondrocytes. The integrated absorbency (IA) value of Col II in group C (10.290 7 +/- 0.499 2) was lower than that in group A (15.377 0 +/- 0.571 8) and higher than that in group B (5.463 2 +/- 0.450 4), showing significant differences (P < 0.05). The IA value of MMP-13 in group C (3.068 6 +/- 0.205 6) was significantly lower than that in group B (6.998 1 +/- 0.329 7, P < 0.05), but there was no significant difference when compared with group A (2.777 5 +/- 0.199 6, P > 0.05). The IA value of beta-catenin in group C (6.611 7 +/- 0.381 8) was lower than that in group B (11.799 9 +/- 0.348 7) and higher than that in group A (4.390 3 +/- 0.551 9), showing significant differences (P < 0.05). The mRNA expression of Col II in group C was significantly higher than those in groups A and B (P < 0.05), the mRNA expression of MMP-13 in group C was significantly lower than that in group B (P < 0.05) but there was no significant difference when compared with group A (P > 0.05). The mRNA expression of j-catenin in group C was significantly lower than that in group B (P < 0.05) and higher than that in group A (P < 0.05).
ALN can protect rat chondrocyte from OA induced by IL-1beta in vitro possibly by upregulating Col II and inhibiting the expression of MMP-13 and beta-catenin in the chondrocytes.
通过观察阿仑膦酸钠（ALN）对白细胞介素1β（IL-1β）诱导的大鼠软骨细胞的影响，探讨ALN治疗骨关节炎（OA）的可行性。
培养15只SD大鼠（1月龄，雌雄不限，体重100 - 150 g）膝关节表面软骨细胞。通过倒置相差显微镜观察软骨细胞，并采用甲苯胺蓝染色和苏木精-伊红（HE）染色进行鉴定。将第3代软骨细胞分为3组：A组用 Dulbecco's改良 Eagle培养基（DMEM）培养5天；B组先用10 ng/mL IL-1β培养2天，再用DMEM培养3天；C组先用10 ng/mL IL-1β培养2天，再用1×10⁶ mol/L ALN培养3天。采用免疫细胞化学和实时荧光定量聚合酶链反应（PCR）检测Ⅱ型胶原（ColⅡ）、基质金属蛋白酶13（MMP-13）和β-连环蛋白的表达水平。
甲苯胺蓝染色证明培养的细胞为软骨细胞。C组ColⅡ的积分吸光度（IA）值（10.290 7±0.499 2）低于A组（15.377 0±0.571 8），高于B组（5.463 2±0.450 4），差异有统计学意义（P<0.05）。C组MMP-13的IA值（3.068 6±0.205 6）显著低于B组（6.998 1±0.329 7，P<0.05），但与A组（2.777 5±0.199 6，P>0.05）比较差异无统计学意义。C组β-连环蛋白的IA值（6.611 7±0.381 8）低于B组（11.799 9±0.348 7），高于A组（4.390 3±0.551 9），差异有统计学意义（P<0.05）。C组ColⅡ的mRNA表达显著高于A组和B组（P<0.05），C组MMP-13的mRNA表达显著低于B组（P<0.05），但与A组比较差异无统计学意义（P>0.05）。C组β-连环蛋白的mRNA表达显著低于B组（P<0.05），高于A组（P<0.05）。
ALN可能通过上调ColⅡ表达、抑制软骨细胞中MMP-13和β-连环蛋白的表达，在体外保护大鼠软骨细胞免受IL-1β诱导的OA损伤。
