Electroacupuncture regulates inflammation, collagen deposition and macrophage function in skeletal muscle through the TGF-β1/Smad3/p38/ERK1/2 pathway

Skeletal muscle injury is one of the most common sports injury, which accounts for similar to 40% of all sports-related injuries among the elderly. In addition, cases of full recovery from treatment are rare. Although electroacupuncture (EA) is an integral aspect of traditional Chinese medicine, the effects of EA on skeletal muscle fibrosis and the possible underlying mechanism remain unclear. To investigate the effect and potential mechanism of EA on skeletal inflammation, collagen deposition and macrophage function, a skeletal muscle injury model was established by injecting 100 mu l cardiotoxin into the anterior tibial muscle of Sprague Dawley rats. The animals were randomly divided into the following three groups: Control, model and EA. The expression of inflammation-related factors (IL-6, IL-4, IL-33, IL-10 and TNF-alpha) were measured using ELISA. H&E staining, Masson's staining and immunohistochemistry (collagen II, Axin2 and beta-catenin) were performed to assess collagen deposition and fibrosis in the muscle tissues. Additionally, immunofluorescence was performed to measure the ratio of M-1 to M-2 macrophages. Western blotting was performed to examine the activity of the TGF-beta 1/Smad3/p38/ERK1/2 pathway. Compared with that in the control rats, the mental state, such as the degree of activity and excitement, of the model rats deteriorated, with clear activity limitations. Compared with those in the model rats, EA-treated rats exhibited improved mental status and activity, reduced levels of IL-6, IL-4 and TNF-alpha, reduced collagen deposition and fibrosis, in addition to increased expression of IL-33 and IL-10. This improvement became increasingly evident with prolonged intervention time. EA also promoted the transformation of macrophages from the M-1 into the M-2 sub-type, where the M-1/M-2 ratio on day 7 was lower compared with that on day 14. Western blotting results showed that compared with that in the model rats, the expression of TGF-beta 1, MMP-2, MMP-7 and the activation of Smad3 and p38 was decreased in EA-treated rats, whilst the activation of ERK1/2 was significantly elevated. In conclusion, EA can inhibit inflammation and collagen deposition whilst promoting the transformation of macrophages from the M-1 into the M(2)sub-type. The underlying mechanism was found to be associated with TGF-beta 1/Smad3/p38/ERK1/2 signaling.

Automated summary

Electroacupuncture (EA) can inhibit inflammation and collagen deposition, promote the transformation of macrophages from M-1 to M-2 subtype, and the underlying mechanism is associated with TGF-beta 1/Smad3/p38/ERK1/2 signaling pathway.