HPLC法同时测定复方茶碱麻黄碱片中4组分的含量

目的:用高效液相色谱法同时测定复方茶碱麻黄碱片中可可碱、盐酸麻黄碱、茶碱和咖啡因的含量。方法:采用Diamonsil C18色谱柱(250mm×4.6 mm,5μm);流动相:甲醇-0.05 mol·L-1磷酸二氢钾溶液-三乙胺(25:75:0.1);流速:0.8 ml·min-1;检测波长:210nm。结果:可可碱进样量在0.06～0.39μg(r=0.999 6)、盐酸麻黄碱进样量在0.03～0.16μg(r= 0.999 6)、茶碱进样量在0.06～0.38μg(r=0.999 6)、咖啡因进样量在0.04～0.23μg(r=0.999 6)范围内呈良好的线性关系。平均回收率分别为100.3%(RSD=0.3%);100.2%(RSD=0.6%);100.2%(RSD=0.6%);100.7%(RSD=0.5%)。结论:本方法简便、准确、重现性好,可用于同时测定复方茶碱麻黄碱片中4组分的含量。     

Theobromine mitigates IL-1β-induced oxidative stress,inflammatory response,and degradation of type II collagen in human chondrocytes

Osteoarthritis is one of the major causes of disability in elderly adults. Chondrocytes are responsible for the formation and remodeling of articular cartilage in joint tissue. The dysfunction of chondrocytes is a significant factor in the development of osteoarthritis. In the current study, we found that theobromine, a constituent of the cacao plant, possesses a preventive effect against interleukin (IL)-1β-induced chondrocyte dysfunction. Theobromine ameliorates IL-1β-induced production of cellular reactive oxygen species (ROS) and inflammatory mediators including cyclooxygenase-2 (COX-2) and prostaglandin E₂ (PGE₂). The presence of theobromine suppresses IL-1β-induced inducible nitro oxide synthase (iNOS) expression and cellular nitro oxide (NO) production. Theobromine also suppresses IL-1β-induced production of the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCP-1), as well as matrix metalloproteinases (MMP)-3 and MMP-13. Additionally, theobromine mitigates IL-1β-induced type II collagen degradation. Mechanistically, we show that theobromine inhibits IL-1β-induced IκBα activation, nuclear factor-κB (NF-κB) protein p65 accumulation, and transfected NF-κB promoter activity, indicating that theobromine suppresses the NF-κB pathway in chondrocytes. Collectively, our study demonstrates that the natural molecule theobromine has a protective effect to counter cytokine-induced chondrocyte dysfunction, implying its beneficial effect in the prevention of osteoarthritis.