来源：风湿病与关节炎,2023,12（11）:6-10,60.

基于Trx2/ASK1信号通路探讨补骨颗粒对软骨细胞凋亡的调控机制（临床研究）

余光书¹，林焱斌¹，王松清¹，吴春玲²

　　【摘　要】目的：观察补骨颗粒对软骨细胞凋亡及对Trx2/ASK1/Caspase-3表达的影响，从而探讨补骨颗粒对软骨细胞凋亡影响机制，为进一步研究补骨颗粒防治骨关节炎奠定实验基础。方法：将20只SD大鼠随机分为空白对照组，补骨颗粒低、中、高剂量组，每组5只。适应性喂养7 d，空白对照组给予蒸馏水2 mL·kg^-1灌胃，补骨颗粒低、中、高剂量组给予补骨颗粒1.54 g·kg^-1、3.08 g·kg^-1、6.16 g·kg^-1灌胃。干预14 d后，抽取血液经离心获得血清。通过酶联免疫吸附试验检测血清中Trx2、ASK1、Caspase-3表达量，采用MTT法检测血清干预后的细胞增殖活性，采用流式细胞仪检测血清干预后的软骨细胞凋亡率。结果：与空白对照组比较，补骨颗粒低、中、高剂量组Trx2、ASK1与Caspase-3含量明显减少，差异均有统计学意义（P < 0.05）；与补骨颗粒中剂量组比较，补骨颗粒低剂量组和高剂量组Trx2与ASK1含量明显增多，补骨颗粒低剂量组Caspase-3含量明显增多，差异均有统计学意义（P < 0.05）。与空白对照组比较，补骨颗粒低、中、高剂量组OD值明显升高（P < 0.05）；补骨颗粒低、中、高剂量组组间比较，差异无统计学意义（P > 0.05）。流式细胞仪检测发现，不加入硝普钠时（对照组1）软骨细胞的凋亡率最低为4.25%，加入硝普钠后（空白对照组1）软骨细胞的凋亡率变为46.86%，补骨颗粒低剂量组1软骨细胞的凋亡率（30.15%）高于补骨颗粒中剂量组1（27.77%）和高剂量组1（27.41%）。结论：补骨颗粒将会调控Trx2表达，并且不同剂量补骨颗粒将会通过Trx2-ASK1解离度调控Caspase-3活化程度而影响软骨细胞的凋亡，这将有助于进一步认识补骨颗粒对骨关节炎防治过程。

　　【关键词】　骨关节炎；补骨颗粒；Trx2/ASK1信号通路；软骨细胞；细胞凋亡；大鼠

Exploring the Regulatory Mechanism of Bugu Keli（补骨颗粒）on Chondrocyte Apoptosis Based on the Trx2/ASK1 Signaling Pathway

YU Guang-shu,LIN Yan-bin,WANG Song-qing,WU Chun-ling

　　【ABSTRACT】Objective:To observe the effect of Bugu Keli（补骨颗粒）on chondrocyte apoptosis and Trx2/ASK1/Caspase-3 expression,in order to explore its mechanism on chondrocyte apoptosis and lay an experimental foundation for further research on the prevention and treatment of osteoarthritis by Bugu Keli.Methods:Twenty SD rats were randomly divided into a blank control group and low,medium,and high dose groups of Bugu Keli,with 5 rats in each group.Adaptive feeding for 7 days,the blank control group was given 2 mL·kg^-1 of distilled water by gavage,while the low,medium,and high dose groups were given 1.54 g·kg^-1,3.08 g·kg^-1,6.16 g·kg^-1 of Bugu Keli by gavage.After 14 days of intervention,blood was extracted and centrifuged to obtain serum.The expression levels of Trx2,ASK1,and Caspase-3 in serum were detected by enzyme-linked immunosorbent assay.Cell proliferation activity was detected by MTT method after serum intervention,and chondrocyte apoptosis rate was detected by flow cytometry after serum intervention.Results:Compared with the blank control group,the contents of Trx2,ASK1,and Caspase-3 in the low,medium,and high dose groups were significantly reduced,with statistical significance（P < 0.05）;compared with the medium dose group,the low dose and high dose group showed a significant increase in Trx2 and ASK1 content,while the low dose group of Bugu Granules showed a significant increase in Caspase-3 content,with statistical significance（P < 0.05）.Compared with the blank control group,the OD values of the low,medium,and high dose groups were significantly increased（P < 0.05）;there was no statistically significant difference between the low,medium,and high dose groups（P > 0.05）.Flow cytometry detection showed that the lowest apoptosis rate of chondrocytes was 4.25% in control group 1 without the addition of sodium nitroprusside.After the addition of sodium nitroprusside,the apoptosis rate of chondrocytes in blank control group 1 increased to 46.86%.The apoptosis rate of chondrocytes in the low dose group 1（30.15%）was higher than that in the medium dose group 1（27.77%）and the high dose group 1（27.41%）.Conclusion:Bugu Keli will regulate the expression of Trx2,and different doses of it will affect the apoptosis of chondrocytes by regulating the degree of Caspase-3 activation through the dissociation of Trx2-ASK1.This will help to further understand the prevention and treatment process of Bugu Keli on osteoarthritis.

　　【Keywords】 osteoarthritis;Bugu Keli（补骨颗粒）;Trx2/ASK1 signaling pathway;chondrocytes;cell apoptosis;rats