Effects of Couplet Medicinals of Angelica Sinensis and Achyranthes Bidentata Modulation of Bmal1/Clock Signaling Pathway on Cartilage Cells Apoptosis in Rats with Knee Osteoarthritis
Objective:To investigate the effects of couplet medicinals of Angelica Sinensis and Achyranthes Bidentata modulation of brain and muscle ARNT-like 1(Bmal1)/clock rhythm regulating protein(Clock)signaling pathway on cartilage cells apoptosis in rats with knee osteoarthritis(KOA).Methods:Anterior cruciate ligament transection(ACLT)was used to construct a rat KOA model,and the rats after successful modeling were randomly divided into model group,positive control group(celecoxib 18 mg/kg),Angelica Sinensis and Achyranthes Bidentata low-dose group(1.08 g/kg),Angelica Sinensis and Achyranthes Bidentata high-dose group(2.16 g/kg),Angel-ica Sinensis and Achyranthes Bidentata+Bmal1 inhibitor group(Angelica Sinensis and Achyranthes Bidentata 2.16 g/kg+Bmal1 inhibitor SR9009 50 mg/kg),with 7 rats in each group;another 7 rats were taken as the sham opera-tion group.The rats in the sham-operation group and model group were injected with saline by gavage or intraperitoneal injection,and the rats in the remaining groups were injected with the corresponding drugs by gavage or intraperitoneal injection once a day for 28 d.After the last administration,the histopathological changes in the cartilage of the knee joints of rats in each group were observed and Mankin score was performed,the levels of interleukin 1β(IL-1β)and IL-6 in ser-um were detected,the chondrocyte apoptosis was detected,the positive expression of Bmal1 protein in cartilage tissue was detected,and the expression of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),and the expression of cleaved cysteinyl aspartate specific proteinase 3(Cleaved caspase 3)apoptosis-related protein expression,and cartilage tissue Bmal1/Clock signaling pathway-related protein and mRNA expression.Results:Compared with the sham operation group,rats in the model group had broken cartilage surfaces,interrupted continuity,disorganized chondrocyte arrangement,and reduced proteoglycan content in cartilage matrix;the serum levels of IL-1β and IL-6 were significantly elevated;the apoptotic chondrocytes were significantly elevated;the percentage of Bmal1 protein-positive cells in the cartilage tissues was significantly lower,the expression of Bcl-2 protein was significantly lower,and the expressions of Bax,Cleaved caspase 3 protein were significantly elevated,as well as Bmal1,Clock protein and mRNA were significantly elevated(P＜0.05).Compared with the model group,the couplet medicinals of Angelica Sinensis and Achyranthes Bidentata pairs of low-and high-dose groups and the positive control group significantly improved the pathological changes in the cartilage tissue of rats,and all of the above indexes were significantly reversed(P＜0.05);the inhibition of the Bmal1/Clock signaling path-way could attenuate the effects of the couplet medicinals of Angelica Sinensis and Achyranthes Bidentata on the improve-ment of apoptosis of chondrocytes.Conclusion:The couplet medicinals of Angelica Sinensis and Achyranthes Bidentata ameliorated cartilage cells apoptosis in KOA rats,and its effect may be related to the activation of the Bmal1/Clock signa-ling pathway.
knee osteoarthritiscouplet medicinals of Angelica Sinensis and Achyranthes Bidentatabrain and muscle ARNT-like 1/clock rhythm regulating protein signaling pathwayapoptosis
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