摘要
目的探讨断藤益母汤（DYD）含药血清对体外培养的类风湿关节炎（rheumatoid arthritis，RA）患者滑膜成纤维细胞（fibrolast-like synoviocytes，FLS）增殖以及TLR4（Toll like receptor 4）mRNA表达的影响和作用机制。方法体外培养RA-FLS，取第3-5代细胞，分别加入来氟米特和DYD高、中、低剂量家兔含药血清，空白组加入正常家兔血清，Mrrr法检测24h、48h、72h后各组合药血清对RA-FLS的影响。取RA-FLS，空白组不加脂多糖（LPS），阳性对照组加LPS20Ixg·mL-1，实验组在加入LPS后分两组，分别加入来氟米特和细胞抑制率最高的DYD含药血清。Reahime-PCR法检测以上4组细胞TLR4mRNA的表达。结果来氟米特和DYD高剂量组合药血清作用48h、72h，二者均能显著抑制RA-FLS增殖（P〈0．05）；对LPS诱导的TLR4的mRNA表达也有显著抑制作用（P〈0．05）；来氟米特和DYD药效相当，两者比较差异无统计学意义。结论断藤益母汤高、中剂量含药血清作用48h、72h后能显著抑制RA-FLS增殖，且高剂量血清可下调LPS诱导的TLR4mRNA表达，这可能是该方治疗RA的机制之一。
Objective To investigate the effects of serum containing Duanteng Yimu Decoction（DYD） on the prolifer ation of fihroblastlike synoviocytes and TLR4（Tolllike receptor 4） expression in rheumatoid arthritis（RA） patients. Methods Synovial specimens obtained from the joint of RA patients. In the first experiment, the third to fifth gen- erations of subcuhured RAFLS were cocultured with teflunomide or serum containing high, moderate and low- dosage DYD. RA-FLS of control group was cultured with the serum of normal rabbit. After 24, 48 and 72 hours, the morphological features of RAFLS were observed under inverted microscope, Proliferation was measured by MTF method. In the second experiment, RAFLS of control group was cultured without lipopolysaccharide （LPS） added, the expeviment group was added with LPS 20 Ixg ·mL-1, and the experiment group was added with leflunomide or DYD- containing serum with the highest cell inhibiting rate after treatment with LPS. RealtimePCR was used for the observation of TLR4 mRNA in RAFLS. Results Compared to the control group, leflunomide and serum containing highdosage DYD obviously inhibited the proliferation of RA-FLS （P 〈 0.05 ） after culturing for 48 and 72 hours, and had obvious inhibitory effect on TLR4 mRNA after stimulation with LPS （P 〈 0.05）. The effect of serum con taining DYD and leflunomide. Conclusion Serum containing high and moderatedosage DYD can inhibit RAFLS proliferation, and serum containing high dosage DYD can downregulate the expression of TLR4 mRNA, which may be one of the therapeutic mechanisms of DYD for RA.
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2012年第2期147-152,共6页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金面上项目(81173634)
广东省自然科学基金自由申请项目(S2011010005660)
