CAJ | 학술논문

背景：透骨消痛胶囊治疗早、中期骨性关节炎有较好疗效，但作用机制尚未完全阐明。小GTP酶Rho能够抑制软骨细胞肥大分化，促进软骨细胞的凋亡。 　　目的：观察透骨消痛胶囊对体外培养凋亡大鼠关节软骨细胞Rac1和Cdc42的影响，并初步探讨其防治骨性关节炎的作用机制。 　　方法：采用4周龄SD大鼠膝关节软骨建立稳定的软骨细胞体外培养体系，采用甲苯胺蓝染色法对第3代软骨细胞进行鉴定。用20μg/L的肿瘤坏死因子α诱导体外培养软骨细胞的凋亡，诱导成功后，给予透骨消痛胶囊(500，100，20 mg/L)孵育24 h后，分别用MTT法检测各组软骨细胞的存活率，用流式细胞仪检测各组软骨细胞的线粒体膜电位情况，Western Blot检测各组软骨细胞Rac1，Cdc42，Bcl-2及Bax蛋白表达。结果与结论：透骨消痛胶囊能够减少肿瘤坏死因子α所致的软骨细胞凋亡，提高线粒体膜电位，提高细胞的存活率，同时能够下调Rac1，Cdc42，Bax蛋白的表达，上调Bcl-2蛋白表达，差异均有显著性意义(P<0.05)。提示透骨消痛胶囊可能通过下调Rac1，Cdc42和 Bax蛋白表达，增加凋亡抑制基因Bcl-2蛋白表达，从而抑制软骨细胞的凋亡，而起到治疗骨性关节炎的疗效。
배경：투골소통효낭치료조、중기골성관절염유교호료효，단작용궤제상미완전천명。소GTP매Rho능구억제연골세포비대분화，촉진연골세포적조망。 　　목적：관찰투골소통효낭대체외배양조망대서관절연골세포Rac1화Cdc42적영향，병초보탐토기방치골성관절염적작용궤제。 　　방법：채용4주령SD대서슬관절연골건립은정적연골세포체외배양체계，채용갑분알람염색법대제3대연골세포진행감정。용20μg/L적종류배사인자α유도체외배양연골세포적조망，유도성공후，급여투골소통효낭(500，100，20 mg/L)부육24 h후，분별용MTT법검측각조연골세포적존활솔，용류식세포의검측각조연골세포적선립체막전위정황，Western Blot검측각조연골세포Rac1，Cdc42，Bcl-2급Bax단백표체。결과여결론：투골소통효낭능구감소종류배사인자α소치적연골세포조망，제고선립체막전위，제고세포적존활솔，동시능구하조Rac1，Cdc42，Bax단백적표체，상조Bcl-2단백표체，차이균유현저성의의(P<0.05)。제시투골소통효낭가능통과하조Rac1，Cdc42화 Bax단백표체，증가조망억제기인Bcl-2단백표체，종이억제연골세포적조망，이기도치료골성관절염적료효。
BACKGROUND:Tougu Xiaotong Capsule has pretty good clinical therapeutic effect on osteoarthritis of early and middle periods. However, the mechanism of Tougu Xiaotong Capsule is not ful y clarified. The RhoA GTPases can regulate chondrocyte apoptosis and hypertrophy. OBJECTIVE:To observe the Tougu Xiaotong Capsule on the expression of Rac1and Cdc42 in tumor necrosis factor-α-induced in vitro cultured rat articular chondrocytes, and to explore its mechanism of action for combating osteoarthritis. METHODS:Knee cartilage of the 4-week-old Sprague-Dawley rats was used to stably establish in vitro culture system of chondrocytes. Passage 3 chondrocytes were identified by toluidine blue staining. Chondrocyte apoptosis was successful y induced by 20μg/L tumor necrosis factor-αand then Tougu Xiaotong Capsule at different dosage (500, 100, 20 mg/L) was given after 24-hour incubation. MTT assay was used to detect cellsurvival, flow cytrometry to measure mitochondrial membrane potential, and western blot assay to determine the protein expression of Rac1, Cdc42, Bax and Bcl-2. RESULTS AND CONCLUSION:Tougu Xiaotong Capsule could reduce tumor necrosis factor-α-induced apoptosis of chondrocytes to improve the survival rate of the cells, and at the same time, could down-regulate the protein expression of Rac1, Cdc42 and Bax and increase the protein expression of Bcl-2 significantly (P<0.05). Tougu Xiaotong Capsule possibly plays a therapeutic efficacy on osteoarthritis by reducing promote apoptosis Rac1, Cdc42 and Bax expression and increasing apoptosis inhibiting gene Bcl-2 expression, thereby to inhibit apoptosis of chondrocytes.