Objective To explore the effects and therapeutic mechanism of electroacupuncture on the levels of polarization markers and inflammatory factors interleukin(IL)-6,IL-4,tumor necrosis factor-alpha(TNF-α),and IL-10 in rats with para-chlorophenylalanine-induced insomnia(PCPA).Methods Fifty healthy specific-pathogen free grade Sprague-Dawley rats,half male and half female,were randomly divided into a blank group(n=10)and a model reserve group(n=40),in which insomnia was induced by intraperitoneal injection of a 500 mg/kg PCPA suspension.Using the random number table method,the 30 successfully modeled rats were divided into three treatment groups of 10 rats/group:model,electroacupuncture,and estazolam.The estazolam group was given estazolam 0.2 mg/(kg·d)by gavage;the electroacupuncture group was given once-daily electroacupuncture at the"Shenmen"and"Sanyinjiao"acupoints,and stimulation at the"Baihui"and"Benshen"acupoints,20 minutes each time,for 7 consecutive days.Following treatment,serum and hypothalamic levels of TNF-α,IL-6,IL-4,and IL-10 were detected using ELISA and Western blot,while immunofluorescence staining was used to detect the presence of Iba-1 in hypothalamic microglia and the co-expression of CD86 and CD163,which are markers for the M1 and M2 subtypes of microglial cells,respectively.Results Compared with the blank group,the model group exhibited prolonged sleep latency(SL)(P＜0.01),shortened sleep duration(ST)(P＜0.05),significantly higher serum and hypothalamic protein levels of IL-6 and TNF-α(P＜0.01),and significantly lower levels of IL-4 and IL-10(P＜0.01).Compared with the model group,the electroacupuncture and estazolam groups exhibited significantly shorter SL(P＜0.01),prolonged ST(P＜0.01),significantly lower serum and hypothalamic protein levels of IL-6 and TNF-α(P＜0.01),and significantly higher IL-4 and IL-10 levels(P＜0.01).IL-6 content was lower in the electroacupuncture group than in the estazolam group(P＜0.05).Compared with the blank group,the model group exhibited significantly enhanced Iba-1/CD86(M1 type)co-expression(P＜0.01)alongside significantly weakened Iba-1/CD163(M2 type)co-expression(P＜0.01).Under electroacupuncture or estazolam intervention,Iba-1/CD86 co-expression was significantly weakened(P＜0.01),and Iba-1/CD163 co-expression was significantly enhanced in the model group(P＜0.05).Conclusions Electroacupuncture effectively improved sleep disturbances in rats,with an underlying mechanism that may involve regulation of microglial polarization,downregulation of pro-inflammatory cytokine levels,upregulation of anti-inflammatory cytokine levels,and alleviation of neuroinflammation,thereby ameliorating sleep.

Objective To explore the effects and therapeutic mechanism of electroacupuncture on the levels of polarization markers and inflammatory factors interleukin(IL)-6,IL-4,tumor necrosis factor-alpha(TNF-α),and IL-10 in rats with para-chlorophenylalanine-induced insomnia(PCPA).Methods Fifty healthy specific-pathogen free grade Sprague-Dawley rats,half male and half female,were randomly divided into a blank group(n=10)and a model reserve group(n=40),in which insomnia was induced by intraperitoneal injection of a 500 mg/kg PCPA suspension.Using the random number table method,the 30 successfully modeled rats were divided into three treatment groups of 10 rats/group:model,electroacupuncture,and estazolam.The estazolam group was given estazolam 0.2 mg/(kg·d)by gavage;the electroacupuncture group was given once-daily electroacupuncture at the"Shenmen"and"Sanyinjiao"acupoints,and stimulation at the"Baihui"and"Benshen"acupoints,20 minutes each time,for 7 consecutive days.Following treatment,serum and hypothalamic levels of TNF-α,IL-6,IL-4,and IL-10 were detected using ELISA and Western blot,while immunofluorescence staining was used to detect the presence of Iba-1 in hypothalamic microglia and the co-expression of CD86 and CD163,which are markers for the M1 and M2 subtypes of microglial cells,respectively.Results Compared with the blank group,the model group exhibited prolonged sleep latency(SL)(P＜0.01),shortened sleep duration(ST)(P＜0.05),significantly higher serum and hypothalamic protein levels of IL-6 and TNF-α(P＜0.01),and significantly lower levels of IL-4 and IL-10(P＜0.01).Compared with the model group,the electroacupuncture and estazolam groups exhibited significantly shorter SL(P＜0.01),prolonged ST(P＜0.01),significantly lower serum and hypothalamic protein levels of IL-6 and TNF-α(P＜0.01),and significantly higher IL-4 and IL-10 levels(P＜0.01).IL-6 content was lower in the electroacupuncture group than in the estazolam group(P＜0.05).Compared with the blank group,the model group exhibited significantly enhanced Iba-1/CD86(M1 type)co-expression(P＜0.01)alongside significantly weakened Iba-1/CD163(M2 type)co-expression(P＜0.01).Under electroacupuncture or estazolam intervention,Iba-1/CD86 co-expression was significantly weakened(P＜0.01),and Iba-1/CD163 co-expression was significantly enhanced in the model group(P＜0.05).Conclusions Electroacupuncture effectively improved sleep disturbances in rats,with an underlying mechanism that may involve regulation of microglial polarization,downregulation of pro-inflammatory cytokine levels,upregulation of anti-inflammatory cytokine levels,and alleviation of neuroinflammation,thereby ameliorating sleep.