Effect of Tougu Xuejie Powder on JAK2/STAT3 Signaling Pathway and Th17/Treg Imbalance in Rheumatoid Arthritis Mice
Objective:To explore the drug safety of Tougu Xuejie Powder and its effect on the imbalance of Janus kinase 2(JAK2)/sig-nal transducer and activator of transcription 3(STAT3)signaling pathway and regulatory T cells(Treg)/T helper cell 17(Th17)in rheumatoid arthritis(RA)mice.Methods:Acute oral toxicity test:30 C57BL/6 mice were randomly divided into a control group and 5 treatment groups,with 5 mice in each group;The mice in the control group were given a gavage with sterile filtered water,while the mice in the treatment group were given a gavage with different doses of Tougu Xuejie Powder(55 mg·kg-1,175 mg·kg-1,550 mg·kg-1,1 750 mg·kg-1,5 000 mg·kg-1)once a day for 14 consecutive days.Calculate the median lethal dose(LD50)and 95％confidence interval of mice using probability weighted regression(Bliss method).Subacute toxicity test:20 C57BL/6 mice were randomly divided into a blank group and 3 treatment groups,with 5 mice in each group;The blank group mice were given a gavage with sterile filtered wa-ter,while the treatment group mice were given a gavage with different doses of Tougu Xuejie Powder(500 mg·kg-1,1 000 mg·kg-1,1 500 mg·kg-1)for 28 consecutive days,and the poisoning symptoms of the mice were observed daily.Select 10 out of 30 C57BL/6 mice randomly as the normal group and inject remaining mice with chicken type Ⅱ collagen(emulsified with Freund's complete adju-vant)and inject the same material another time on the 21st day to establish an arthritis model.On the 27th day,the successfully mod-eled mice were randomly divided into a model group and a Tougu Xuejie Powder group(1 500 mg·kg-1),with 10 mice in each group.They were orally administered for 5 weeks,once a day.Flow cytometry was used to detect Treg and Th17 in the spleen,and West-ern blot was used to detect the expression levels of JAK2,p-JAK2,STAT3 and p-STAT3 in synovial tissue.ELISA was used to detect tumor necrosis factor alpha(TNF-α),Interleukin-1 β(IL-1 β),Interleukin-17(IL-17)and interleukin-10(IL-10)levels in synovial tissue,and arthritis score(AS)was performed.Results:The LD50 of Tougu Xuejie Powder is 3 878 mg·kg-1,with a 95％confidence interval of 1 470-5 498 mg·kg-1.No death or obvious toxic symptoms were observed in the control group and each dose group of mice treated with Tougu Xuejie Powder.Compared with that of the normal group,the percentage of Treg in the spleen of the model group mice decreased and the percentage of Th 17 increased significantly(P＜0.01).Compared with that of the model group,the percentage of Treg in the spleen of mice in the group treated with Tougu Xuejie Powder increased,while the percentage of Th17 de-creased significantly(P＜0.01).Compared with that of the normal group,TNF-α,IL-1β,IL-17 content in synovial tissue of model group mice increased while the content of IL-10 decreased significantly(P＜0.01).Compared with that of the model group,TNF-α,IL-1β,IL-17 content in synovial tissue of Tougu Xuejie Powder group mice decreased while the content of IL-10 increased signifi-cantly(P＜0.01).Compared with that of the normal group,the expression levels of JAK2,p-JAK2,STAT3 and p-STAT3 increased in the synovial tissue of the model group mice significantly(P＜0.05).Compared with that of the model group,the expression levels of JAK2,p-JAK2,STAT3 and p-STAT3 in the synovial tissue of mice treated with Tougu Xuejie Powder decreased significantly(P＜0.05).Conclusion:Tougu Xuejie Powder can relieve symptoms in RA mice,the mechanism of which may be related to inhibiting the ac-tivation of JAK2/STAT3 signaling pathway,regulating Th17/Treg balance and inhibiting inflammatory response.
万方数据
维普
