Mechanism of effects of pressing Pishu(BL20)and Weishu(BL21)on alleviating peripheral inflammation in rats with chronic fatigue syndrome based on the α7nAChR/NF-κB pathway
Objective To observe the effects of pressing Pishu(BL20)and Weishu(BL21)on the fatigue state,locomotor behaviors,peripheral inflammation,and α 7 nicotinic acetylcholine receptor(α7nAChR)/nuclear factor-KB(NF-κB)signaling expression of chronic fatigue syndrome(CFS)model rats,and to explore the mechanism of effect of this therapy on the alleviation of peripheral inflammation in CFS rats.Methods A total of 32 SD rats were randomized into blank group(n=8)and modeling group(n=24).In the modeling group,a CFS model was established by forced weight-bearing swimming combined with chronic stress stimulation.After successful modeling,the rats in the modeling group were randomly subdivided into model,acupoint-pressing,and α7nAChR agonist groups,with eight rats in each group.The blank and model groups were injected intraperitoneally with an equal volume of saline;the acupoint-pressing group was subjected to the bilateral press on Pishu(BL20)and Weishu(BL21)using a homemade pressing stimulator for 20 min each time as well as intraperitoneal injection of an equal volume of saline;the α7nAChR agonist group was injected intraperitoneally with the α7nAChR agonist PNU-282987(2.4 mg/kg per time).Each group was intervened once daily for continuous 14 days.The semi-quantitative score of the general condition,swimming time to exhaustion,and exercise distance in the open-field test were recorded in each group;the spleen index and thymus index were calculated;the content of tumor necrosis factor-α(TNF-α),interleukin-1 β(IL-1β),and interleukin-6(IL-6)in rat serum were checked using ELISA;the protein expression levels of α7nAChR,NF-κB p65,NF-κB p-p65,and TNF-α in the rat spleen tissue were examined by Western blot;the mRNA expression levels ofα7nAChR,NF-κB p65,and TNF-α were determined by qPCR.Results Compared with the blank group,the semi-quantitative score of the general condition,serum content of TNF-α,IL-1 β,and IL-6,as well as the protein expression levels of NF-κB p-p65,NF-κB p65,and TNF-α and the mRNA expression levels of NF-κB p65 and TNF-α in the spleen tissue increased in the model group(P＜0.01);the swimming time to exhaustion and exercise distance in the open-field test were shortened,and the spleen index,thymus index,and the α7nAChR protein and mRNA expression levels in the spleen tissue decreased(P＜0.05,P＜0.01).Compared with the model group,the semi-quantitative scores of the general condition,serum content of TNF-α,IL-1β,and IL-6,as well as the protein expression levels of NF-κB p-p65,NF-κB p65,and TNF-α and the mRNA expression levels of NF-κB p65 and TNF-α in the spleen tissue were reduced in both acupoint-pressing and α7nAChR agonist groups(P＜0.05,P＜0.01);the swimming time to exhaustion and exercise distance in the open-field test were longer,and the spleen index,thymus index,and the α7nAChR protein and mRNA expression levels in the spleen tissue were higher(P＜0.05,P＜0.01).Compared with the acupoint-pressing group,the serum content of TNF-α,IL-1 β,and IL-6,as well as the protein expression levels of NF-κB p-p65,NF-κB p65,and TNF-α and the mRNA expression levels of NF-κB p65 and TNF-α in the spleen tissue were lower in the α7nAChR agonist group(P＜0.01),while the α7nAChR protein and mRNA expression levels in the spleen tissue were elevated(P＜0.05,P＜0.01).Conclusion Pressing Pishu(BL20)and Weishu(BL21)can effectively relieve fatigue,improve locomotor behaviors,enhance immune level,and alleviate peripheral inflammation in CFS rats,which may be related to the activation of the expression of α7nAChR,thereby inhibiting the activity of the downstream inflammatory pathway of NF-κB.
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