Exploring Mechanism of Intestinal Inflammation Regulation by Acupuncture in Rats with Spleen Deficiency Based on miRNA-29c/LIF Axis
Objective To observe the mechanism of miRNA-29c/leukemia inhibitory factor(LIF)axis regulation by acupunc-ture to improve colonic inflammation in rats with spleen deficiency and explore the modern mechanism of"strengthening spleen and benefiting Qi"acupuncture therapy to improve the symptoms of spleen deficiency and loose stool.Methods Sixty SD male rats were divided into normal group,model group,non-acupoint group and acupuncture group by random number table method,with 15 rats in each group.Except for the normal group,the remaining three groups were used to establish the rat model of spleen deficiency syndrome by the compound method of overwork and irregular diet.After successful modelling,the acupuncture group was treated with electro-acupuncture at the"Zusanli(ST36)"and"Pishu(BL20)",while the non-acupoint group was treated with electro-acupuncture at the tail tip as a control,once a day for 20 min for a total of 14 d.After treatment,colon tissues and whole blood were collected from the rats.The expressions of miRNA-29/LIF axis-related genes in rat colon tissues were detec-ted by qPCR.The expressions of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in rat colon tissues were de-tected by Western blot.The contents of interleukin-1 β(IL-1 β)and nterleukin-18(IL-18)in colon tissues were detected by ELISA.The pathological morphological changes in rat colon tissues were observed by HE staining.The apoptosis of colon tis-sues in each group was observed by TUNEL staining.Flow cytometry was used to observe the apoptotic status of peripheral blood mononuclear cells(PBMC cells).Results Compared with those of the normal group,the expression of LIF in colon tissues of rats in the model group decreased significantly(P＜0.05),while the expressions of miRNA-29,IL-1 β,IL-6,I L-8 and TNF-αmRNA and IL-6 and TNF-α protein increased(P＜0.05).Compared with those of the model group,the expression of LIF in the colon tissue of rats in the acupuncture group was significantly increased(P＜0.05),and the expressions of miRNA-29c,IL-1 β,IL-6,IL-8,TNF-α mRNA and IL-6 and TNF-α protein were decreased(P＜0.05),and no significant difference was found in the non-acupoints group(P＞0.05).Compared with those of the normal group,the expressions of pro-inflamma-tory factors IL-1 β and IL-18 in the colon tissue of rats in the model group were significantly increased(P＜0.05).The ex-pressions of pro-inflammatory factors IL-1 β and IL-18 were significantly increased in the colon tissue of rats in the model group compared with those in the normal group(P＜0.05),and decreased in the colon tissue of rats in the acupuncture group compared with those in the model group(P＜0.05).In addition,HE staining results showed that acupuncture treatment could ef-fectively reduce inflammatory cell infiltration and loss of cupped cells in colonic mucosal tissues.TUNEL staining results showed that acupuncture could inhibit miRNA-29c/LIF axis,improve colonic inflammation and inhibit apoptosis in colonic mucosal cells in rats with spleen deficiency.The flow cytometry results showed that compared with that in the normal group,PBMC apoptosis in rats in the model group and non-acupoint group was significantly higher in the model group and non-acupoint group compared with that in the normal group(P＜0.05),and decreased in the acupuncture group compared with that in the model group(P＜0.05).Conclusion Electro-acupuncture at"Zusanli(ST36)"and"Pishu(BL20)"can regulate the miRNA-29c/LIF axis and inhibit the expressions of intestinal inflammatory and pro-inflammatory mediators,thereby inhibiting the apoptosis of blood and colonic mucosal cells and improving the immunity of the body,and improving the symptoms of spleen deficiency and loose stool.
万方数据
维普
