补肾强督方对强直性脊柱炎患者外周血单个核细胞产生MMP-9和TIMP-1的影响

摘要

目的:为探讨基质金属蛋白酶在强直性脊柱炎炎性骨破坏中的作用和补肾强督方治疗强直性脊柱炎的作用机制,比较强直性脊柱炎患者外周血单个核细胞(PBMC)产生基质金属蛋白酶9(MMP-9)及基质金属蛋白酶组织抑制因子1(TIMP-1)与健康对照者之间的差别.并研究补肾强督方治疗前后两者的变化.方法:2005年3月至2006年3月活动期强直性脊柱炎患者30例,其中男27例,女3例;年龄16～15岁,平均(30.8±8.8)岁;病程0.5～10年.经补肾强督方治疗3个月后,做自身前后对照,并设立健康对照组20例,常规分离血清和PBMC,将PBMC用PHA/PMA刺激后收集上清,应用RT-PCR检测PBMC的MMP-9和TIMP-1的mRNA表达水平,应用EUSA检测血清和细胞上清中MMP-9和,TIMP-1的含量.结果:与健康对照组相比,患者治疗前血清中MMP-9和TIMP-1浓度明显升高,患者治疗后与治疗前相比MMP-9和TMMP-1浓度显著降低.经PHA/PMA刺激后,患者治疗前的PBMC表达MMP-9和TIMP-1 mRNA水平明显上调,细胞上清液中MMP-9和TIMP-1量明显升高,与健康对照组相比差异有统计学意义.患者治疗后PBMC表达MMP-9和TIMP-1 mRNA水平明显下调,细胞上清液中MMP-9和,TIMP-1含量均显著下降,与治疗前相比差异有统计学意义.结论:强直性脊柱炎活动期患者的PBMC表达和释放MMP-9和TIMP-1增强.补肾强督方可以显著降低强直性脊柱炎活动期患者MMP-9和TIMP-1的产生.%Objective: In order to investigate the roles of metalloproteinase in inflammatory bone destruction in ankylosing spondylitis (AS) ,and analyze the mechanism of preventing inflammatory bone destruction of Bushen Qiangdu decoction (BSQDD) in AS cases.Comparisons were made on the expressions of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) by peripheral blood mononuclear cells (PBMC) between AS patients and healthy controls.The effect of BSQDD was investigated on the expression and of MMP-9 and TIMP-1 produced by PBMC in AS patients.Methods:From March 2005 to March 2006,30 active AS cases of Kidney-asthenia, Du-cold and blood-stasis syndrome were selected as patients group in the China-Japan Friendship Hospital.There are 27 male patients and 3 female patients.The age range is from 16 to 45 ,averaging (30.8±8.8) years.Disease duration is from 0.5 to 10 years.Cases received three-month BSQDD treatment were considered as the treatment group.Twenty healthy persons were included in the control group.Serum and PBMC were separated.The PBMC were stimulated by PHA and PMA,and the supernatant was collected.The mRNA expression of MMP-9 and TIMP-1 in PBMC was analyzed by RT-PCR.The content of MMP-9 and TIMP-1 in serum and culture supernatant of PBMC were detected by ELISA.Results: Compared with health control group, the serum concentration of MMP-9 and TIMP-1 in patients group before treatment increased (P＜0.01 ,P＜0.05) ,but the level of MMP-9 and TIMP-1 in the serum of patients after treatment decreased compared with pre-treatment cases (P＜0.05).Furthermore,compared with health control group,PBMC of patients group before treatment expressed higher levels of MMP-9 and TIMP-1 both on transcript level and at protein level (P＜0.01 ,P＜0.05), and the expression levels of MMP-9 and TIMP-1 in PBMC in patients after treatment both on transcript level and at protein level was down-regulated compared with pre-treatment (P＜0.01 ,P＜0.05).Conclusion:PBMC of AS patients had a higher potential capacity for MMP-9 and TIMP-1.BSQDD possibly prevented inflammatory bone destruction of AS through inhibiting production of MMP-9 and TIMP-1 produced by PBMC.