Electroacupuncture and Moxibustion Regulate Hippocampus Glia and Mitochondria Activation in DSS-Induced Colitis Mice

Abstract

Objectives: To study the influence of electroacupuncture (EA) and moxibustion on the hippocampus astrocyte and microglia activation in the ulcerative colitis model and to evaluate the mitochondria activity.

Methods: 2.5% dextran sodium sulfate-induced colitis mice were treated by EA or moxibustion. Intestinal pathological structure was observed by hematoxylin and eosin (H&E) staining; the expression of GFAP or S100b (markers for astrocyte), Iba-1 (a marker for microglia), and Mitofilin (a marker for mitochondria) in hippocampus was detected by immunofluorescence staining or western blot.

Results: The results demonstrated that both EA and moxibustion could improve the morphology of distal colonic mucosal epithelia in DSS-induced colitis mice. Expression of GFAP in the hippocampus was significantly increased after EA or moxibustion treatment. The effects were further supported by WB results. Meanwhile, expression of mitofilin in the hippocampus CA1 and CA3 regions showed the same trend as that of GFAP. Expression of Iba-1 in the hippocampus showed no significant difference after EA or moxibustion treatment, while the state of microglia changed from resting in control mice to activated state in colitis mice.

Conclusion: EA and moxibustion were able to modulate the activation of astrocyte, microglial, and mitochondria in the hippocampus area in the colitis model.

Figure 1

Schematic diagram for the experimental procedure. Horizontal black arrow line represents the whole experimental procedure and the key time points. The solid arrow represents the oral administration of 2.5% DSS in drinking water to induce colitis until the 14^th day, and the dotted arrow represents drinking clean water as a control group. The EA or MOX treatment procedure applied to mice was started from the 5^th day of DSS water administration. DSS, dextran sodium sulfate; EA, electroacupuncture; MOX, moxibustion.

Figure 2

Morphological changes in different groups. Leukocyte infiltration into the mucosa and damage to the colon wall were evident in the DSS-induced colitis group (H&E, 200x). (a) Con, control group; (b) DSS, DSS-induced colitis group; (c) DSS + MOX, moxibustion treatment group; (d) DSS + EA, electroacupuncture treatment group.

Figure 3

EA and moxibustion promote astrocyte activation. Labeling (a) and analysis of astrocytes in the hippocampus CA1 (b), CA3 (c), and DG (d) regions of UC mice. Green, GFAP (a marker for astroglia); blue, DAPI nuclear staining (n = 5/group). White arrow highlights the activated astrocyte. Scale bar, 50 μm. (e, f) Western blot (e) and analysis (f) showing the downregulation of S100b (a marker for astroglia) expression in the hippocampus in the DSS-induced colitis group and upregulation after EA and moxibustion treatment (n = 3/group). GFAP, glial fibrillary acidic protein.

Figure 4

EA and moxibustion modulate microglia activation. Labeling (a) and analysis (b–d) of microglia in the hippocampus CA1 (b), CA3 (c), and DG (d) regions of UC mice. Green, Iba-1 (a marker of activated microglia); blue, DAPI nuclear staining (n = 5/group). Scale bar, 50 μm. White arrow highlights the state change of microglia, from resting in control mice (faint Iba-1 staining) to activation in UC mice (DSS-induced; strong Iba-1 staining) and also improvement after EA and moxibustion treatment. Iba1, ionized calcium binding adapter molecule 1.

Figure 5

EA and moxibustion promote mitochondria activation. Representative labeling (a) and analysis (b–d) of mitochondria in the hippocampus CA1 (b), CA3 (c), and DG (d) regions of UC mice. Mitofilin (a marker of activated mitochondria); blue, DAPI nuclear staining (n = 5/group). Scale bar, 50 μm. Below: enlarged views of the white frames in the above panels highlight the cytoplasmic positive expression of mitochondria in the DG region of hippocampus.