温经通络汤含药血清通过PI3K/AKT信号通路对IL-1β诱导软骨细胞凋亡的影响

Effect of Wenjingtongluo decoction drug contained serum on IL-1β induced chondrocyte apoptosis through PI3K/AKT signaling pathway

目的 探索温经通络汤含药血清对IL-1β诱导的软骨细胞凋亡的影响及潜在分子机制。方法 取传代培养的第2代小鼠软骨细胞,随机分为空白对照组(C组)、10ng/L IL-1β诱导模型组(M组)、10ng/L IL-1β+不同浓度含药血清组(10%、15%、20%),分别处理细胞24h后采用流式细胞术检测细胞凋亡率;蛋白印迹分析BAX、Bcl-2、Caspase-3、Caspase-9、p-PI3K、PI3K、p-AKT及AKT蛋白的表达水平。结果 与对照组相比,10ng/L IL-1β可诱导软骨细胞的凋亡(P<0.05);与模型组比,10%、15%及20%温经通络汤含药血清组软骨细胞的凋亡率显著下降,其中20%温经通络汤含药血清组效果最为显著(P<0.05);与对照组相比,模型组BAX、Caspase-3、Caspase-9的表达明显增高(P<0.05),Bcl-2、p-PI3K、PI3K、p-AKT及AKT表达明显下降(P<0.05);不同浓度的温经通络汤含药血清可降低BAX、Caspase-3、Caspase-9表达,增加Bcl-2、p-PI3K、PI3K、p-AKT及AKT表达(P<0.05)。结论 温经通络汤可以通过激活PI3K/AKT信号通路而抑制IL-1β诱导的软骨细胞凋亡,从而起到预防骨性关节炎发生发展的作用。

Objective To explore the effect of Wenjingtongluo decoction drug contained serum on IL-1 β induced chondrocyte apoptosis and its potential molecular mechanism.Methods The chondrocytes of the second generation of mice were randomly divided into blank control group(group C) and 10ng/L IL-1β induction model group(group M), 10ng/L IL-1β+different concentrations of Wenjingtongluo decoction drug contained serum groups(10%,15%,20%).Cells were treated for 24 hours, and the apoptosis rate was detected by flow cytometry;The expression levels of Bax, Bcl-2, Caspase-3, caspase-9, p-pi3k, PI3K, p-Akt and Akt proteins were analyzed by Western blot. The experimental scheme was approved by the animal experiment ethics committee of Nanjing University of traditional Chinese medicine.Results(1) compared with the control group, IL-1β can induce chondrocyte apoptosis(P< 0.05);(2)Compared with the model group, the apoptosis rate of chondrocytes in 10%, 15% and 20% Wenjingtongluo decoction drug contained serum groups decreased significantly, and the effect of 20% Wenjingtongluo decoction drug contained serum group was the most significant(P< 0.05);(3)Compared with the control group, the expressions of Bax, caspase-3 and caspase-9 increased significantly(P<0.05),while the expressions of Bcl-2, p-pi3k, PI3K, p-Akt and Akt decreased significantly(P< 0.05) in model group;Different concentrations of Wenjingtongluo decoction drug contained serumcould reduce the expression of Bax, caspase-3 and caspase-9, and increase the expression of Bcl-2, p-PI3K, PI3K, p-AKT and AKT(P<0.05).Conclusion The results show that Wenjingtongluo decoction may inhibit IL-1β induced chondrocyte apoptosis by regulating PI3K/Akt signal pathway, so as to prevent the occurrence and development of osteoarthritis.