健膝壮骨方对大鼠骨性关节炎软骨细胞凋亡及其调控基因P53、Bcl-2表达作用的影响

Effect of Jianxi Zhuanggu Formula on Apoptosis of Cartilage Cells and Regulating Gene Expressions of P53 and Bcl-2 in Rats with Knee Osteoarthritis

目的:明确健膝壮骨方对骨性关节炎模型大鼠的药效作用,初步探讨其治疗骨性关节炎的作用机理,为将其研发为中药新药提供实验基础。方法:将60只大鼠随机分成正常组(A)10只和手术组50只,将手术组造成膝骨性关节炎模型。造模方法:采用改良Hulth法制备大鼠膝骨性关节炎模型,并按随机数字法分为模型对照组(B)、壮骨关节丸阳性对照组(C)、健膝壮骨方高剂量组(D)、健膝壮骨方中剂量组(E)、健膝壮骨方低剂量组(F)5组。于造模7天后,高、中、低剂量给药组每日灌服健膝壮骨方药液(按每100g体重灌胃2mL),剂量分别相当于生药8g、4g、2g;阳性对照组每日灌服壮骨关节丸水溶液(按每100g体重灌胃2mL),相当于成药2 g;正常组和模型组分别灌服等量生理盐水。分别在治疗后1个月和2个月处死一半动物,取股骨内侧髁软骨,检测各组软骨细胞凋亡及其调控基因P53、Bcl-2的表达情况。结果:健膝壮骨方高、中、低剂量组及壮骨关节丸阳性对照组均在一定程度上转变了软骨细胞凋亡亢进的趋势,在下调P53过度表达及调高受抑制的Bcl-2蛋白表达方面均优于模型对照组,治疗组与阳性药物对照组之间有显著性差异(P﹤0.05)。结论:软骨细胞凋亡是导致骨性关节炎发病的重要机制之一,健膝壮骨方能较好地转变软骨细胞凋亡亢进的趋势,这种作用可能与影响P53、Bcl-2的表达有关,从而在一定程度上保护软骨细胞,对骨性关节炎的发生和发展有预防和延缓作用。

Objective ：To clear the cure effect of Jianxi Zhuanggu tormula on rats with knee osteoarthritis and explore the treatment mechanism. To provide experimental basis for the development of new drugs of TCM. Methods ：60 rats were divided into normal group A （ 10 eases）and surgical group （50 eases）randomly. Rat knee osteoarthritis model was induced with the improved method of Hulth. Surgical method and the model rats were randomly divided into control group B, Zhuangguguanjie Pill positive control group C, high dose of Jianxi Zhuanggu group D, moderate dose of Jianxi Zhuanggu group E and low dose of Jianxi Zhuanggu group F. 7 days after modeling, rats in high, moderate and low dose of Jianxi Zhuanggu groups were given drugs respectively every day（2mL/100g weight） ,dosages was equal to 8g,4g, and 2g crude drugs respectively ,while rats in positive control group were given Zhuangguguanjie pills（ 2mL/100g weight） , dosage was equal to 2g crude drug. Normal group and the model group took equal amounts of saline solution filling respectively. Half rats were killed 1 and 2 months later. The cartilage of lateral femoral condyle was taken to measure apoptosis of cartilage cells and regulation of expressions of P53 and Bel - 2. Results： Experiment group demonstrated the ability of Jianxi Zhuanggu formula and Zhuangguguanjie pills converted the apoptosis trend of the cartilage cells compared to other groups. It was superior to model group in down regulating the hyperexpression of gene P53 and turning up the suppressed Bcl - 2 expression. There was a significant difference （ P 〈 0. 05 ）. Conclusions ： Cartilage apoptosis is one of the important patho- genesis of osteoarthritis. Janxi Zhuanggu formula can convert the apoptosis trend of the cartilage cells. This may be relat- ed to the influence of gene expressions of P53 and Bcl - 2 which protect cartilage cells to some extent and prevent devel- opment of osteoarthritis.