Objective To study the effect mechanism of Xiaoyanzhitong ointment on lipopolysaccharide-induced C2C12 myofibers. Methods C2C12 skeletal muscle cells were cultured in vitro and differentiated into mature muscle tubes with 2% horse serum. The blank group, model group, diclofenac sodium group and Xiaoyanzhitong group were set. LPS was added on the non-blank groups to explore the optimal inflammatory concentration and duration of LPS on C2C12, and to replicate the optimal inflammatory cell model of skeletal muscle in vitro, thus to explore the optimal intervention concentration of Xiaoyanzhitong ointment and diclofenac sodium on C2C12 cell inflammation model. Cell activity was detected by CCK-8 method. The concentrations of IL-1β and IL-6, SOD and MDA content in LPS treated C2C12 cells were detected by ELISA after the addition of Xiaoyanzhitong ointment. Results ① The results of differentiation experiment showed that C2C12 differentiation was obvious on the 4th~5th day. ② C2C12 inflammatory model results showed that the inflammatory reaction of C2C12 cells was the most obvious after 24h treatment with 1 μg/ml LPS, and CCK-8 result showed there was no significant change in cell activity. ③ The results of drug concentration exploration showed that the inflammatory reaction of C2C12 was the lightest under the treatment of 1.25 mg/ml Xiaoyanpain-relieving ointment solution and 6.25 μg/ml diclofenac sodium solution for 24 h, and CCK-8 result showed no significant change in cell activity. ④ Compared with model group, 1.25 mg/ml Xiaoyanzhitong ointment significantly inhibited IL-1β level and MDA level, and increased SOD level in LPS-induced C2C12. Compared with diclofenac sodium group, There was no significant difference of 1.25 mg/ml Xiaoyanzhitong ointment in the inhibition of IL-1β level in LPS-induced C2C12 cells (P=0.188), but there were significant differences in the enhancement of SOD level (P=0.0001) and inhibition of MDA level (P=0.014). Conclusions The composition of Xiaoyanzhitong ointment can reduce the levels of IL-1β, IL-6 and MDA in C2C12 cells under LPS administration, and increase the level of SOD, which may be related to the reduction of tissue inflammation and the alleviation of oxidative stress damage.
Key words
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C2C12 /
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LPS /
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Inflammation /
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Oxidative stress
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