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The Effect and Mechanism of Ligustilide Cycloprolactam on Inhibiting Apoptosis and Inflammation of Rat Chondrocytes

CHEN Xin^1,2, WANG Xinyue³, LI Yuanzhen², QI Xin¹, XI Fangqin², LI Ning^1,2, LIU Junxi⁴, XIE Xingwen^1,2*, LIU Jianjun¹*

(¹Affiliated Hospital of Gansu University of Traditional Chinese Medicine, Lanzhou 730000, China; ²Gansu University of Traditional Chinese Medicine, Lanzhou 730020, China; ³Beijing University of Chinese Medicine, Beijing 100105, China; ⁴Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, China)

Abstract:

This study investigated the effect and mechanism of LIGc (ligustilide cycloprolactam) on apoptosis and inflammation of rat chondrocytes induced by IL-1β (10 ng/mL). Rat chondrocytes were divided into Control group, IL-1β group and LIGc high-dose, medium-dose and low-dose groups. Chondrocyte inflammation model was established by IL-1β induction in other groups except Control group. LIGc high-dose, medium-dose and low-dose groups were treated with 0.4, 0.2 and 0.1 μmol/mL LIGc for 24 h respectively. The effect of LIGc on the activity of rat chondrocytes was detected by CCK-8. The apoptosis of rat chondrocytes was observed by Hoechst 33258 staining. The protein expressions of Bcl-2, Caspase-3, TLR4 and NF-κB p65 were detected by Western blot. The expression levels of COX-2 and HMGB1 mRNA were detected by RT-qPCR. It was found that different concentrations of LIGc had no significant effect on the survival rate of rat chondrocytes. Compared with the Control group, the apoptosis of IL-1β group was significantly increased. The expression level of Caspase-3 protein was significantly increased (P<0.01), and the expression level of Bcl-2 protein was significantly decreased (P<0.01). The expression levels of COX-2 and HMGB1 genes were significantly increased (P<0.01); the expression levels of TLR4 and NF-κB p65 protein were significantly increased (P<0.01). After LIGc intervention, cell viability was significantly increased, cell apoptosis was inhibited, and the expression level of Caspase-3 protein was significantly decreased (P<0.01), the expression level of Bcl-2 protein LIGc (0.4, 0.2 μmol/mL) was significantly increased (P<0.01); the expression levels of COX-2 and HMGB1 genes were significantly decreased (P<0.01). The expression levels of TLR4 and NF-κB p65 protein were significantly decreased (P<0.01). Therefore, LIGc can inhibit the apoptosis and inflammatory response of chondrocytes induced by IL-1β, and its mechanism may be related to the inhibition of TLR4/NF-κB pathway

CSTR: 32200.14.cjcb.2023.05.0009