Effect and mechanism of L-3-n-butanobenzene on experimental autoimmune encephalomyelitis
Objective To explore the therapeutic effect and its potential mechanism of L-3-n-butylphthalide(NBP)on experimental autoimmune encephalomyelitis(EAE)in mice based on the RhoA/ROCK signaling pathway.Methods Totally 32 female C57BL/6 mice were randomly divided into four groups:control group,EAE group,low-dose NBP group(L-NBP),and high-dose NBP group(H-NBP),with 8 mice in each group.C57BL/6 mice were immunized with myelin oligodendrocyte glycoprotein 35-55(MOG35-55)antigen emulsion to induce EAE model.After the modeling,the mice in L-NBP group and H-NBP group were intraperitoneally injected with 3.25,6.5 mg/(kg·d)NBP for 28 d,respectively.The body weight of mice was measured every seven days from the day of immuniza-tion.The neurological function scores were recorded daily.Histopathological changes of spinal cord tissue were observed by HE staining and Luxol fast blue(LFB)staining.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of IL-10,IL-1 β,IL-6,and IL-18 in peripheral blood.Real-time PCR was used to detect the expressions of inflammatory factors TNF-α,TNFR1,and IL-1 β in spinal cord tissue.Western blot was used to detect the expressions of RhoA,ROCK Ⅰ,and ROCK Ⅱ proteins.Results Compared with control group,the body weight was decreased,the neurological function score and the histopathological score were increased,IL-10 level was decreased,IL-18,IL-6,and IL-1 β levels were increased,the mRNA levels of TNF-α,TNFR1and IL-1 β were increased,and the protein expressions of RhoA,ROCK Ⅰ,and ROCK Ⅱ were increased in EAE group(all P＜0.05).Compared with EAE group,the body weigh was increased,the incubation period was prolonged,the peak period was delayed,the neurological function score and the histopathological score were decreased,IL-10 level was increased,IL-1 β,IL-6,and IL-18 levels were decreased,the mRNA levels of TNF-α,TNFR1,and IL-1 β were decreased,and the protein expressions of RhoA,ROCK Ⅰ,and ROCK Ⅱ were decreased in L-NBP group and H-NBP group(all P＜0.05).Compared with L-NBP group,the body weigh was increased,the incubation period was prolonged,the peak period was delayed,the neurological function score and the histopathological score were decreased,IL-10 level was increased,IL-1β,IL-6,and IL-18 levels were decreased,the mRNA levels of TNF-α,TNFR1,and IL-1 β were decreased,and the protein expressions of RhoA and ROCK Ⅱ were decreased in H-NBP group(all P＜0.05).Conclusion NBP can alleviate the peripheral and central inflammatory reactions in EAE mice,thereby improving the neurological symptoms of EAE,which may be achieved by inhibiting the RhoA/ROCK signaling pathway.
L-3-n-butylphthalidemultiple sclerosisautoimmune encephalomyelitisRhoA/ROCK signal pathwayneuroin-flammationC57BL/6 mice
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维普
