Electroacupuncture at ST-36 Protects Interstitial Cells of Cajal via Sustaining Heme Oxygenase-1 Positive M2 Macrophages in the Stomach of Diabetic Mice

Abstract

Background. Electroacupuncture (EA) at ST-36 has been reported to improve delayed gastric emptying and protect the networks of ICC in diabetic models. However, the mechanisms of the effects of EA are still unclear. The purpose of this study was to investigate whether the HO-1 positive M2 macrophages participate in the protective effects of EA for the ICC networks. Methods. Male C57BL/6 mice were randomized into five groups: the normal control group, diabetic group (DM), diabetic mice with sham EA group (SEA), diabetic mice with low frequency EA group (LEA), and diabetic mice with high frequency EA group (HEA). ICC network changes were detected by Ano1 immunostaining. F4/80 and HO-1 costaining was used to measure HO-1 positive macrophage expression. Western blot and PCR methods were applied to monitor HO-1, IL-10, and macrophage markers, respectively. The serum MDA levels were detected by a commercial kit. Results. This study presents the following results: (1) Compared with the control group, ICC networks were severely disrupted in the DM group, but no obvious changes were found in the LEA and HEA groups. (2) Many HO-1 positive macrophages could be observed in the LEA and HEA groups, and the expression of HO-1 was also markedly upregulated. (3) The IL-10 expression was obviously upregulated in the LEA and HEA groups. (4) The serum MDA levels were decreased in the real EA group. (5) When compared to the DM group, the expression of CD163 and Arg-1 was increased in the LEA and HEA groups, but the iNOS expression was decreased. Conclusion. The protective effects of EA on the networks of ICC may rely on the HO-1 positive macrophages to mediate anti-inflammatory and antioxidative stress effects.