| Objective: Effect of different doses of Bushen Jiedu Tongluo Formula on Wistar in rats withEAE clinical manifestations, CNS pathological changes and pathogenesis of IL-6andTGF-β, to explore its role in the prevention and treatment of MS and mechanisms of action,in order to explore the multiple sclerosis disease to new therapeutic approaches to provide ameaningful basis of experimental evidence.Methods:(1) EAE model:The guinea pig spinal cord homogenate (GPSCH), completeFreund’s adjuvant (CFA), pertussis vaccine(PV) puree liquid as antigens, sensitized femaleWistar rats to establish acute EAE model, rats were observed for clinical signs, body weightchanges and nerve recording funciton score.(2) Animal grouping and Drug Administration:90female Wistar rats were randomlydivided into6groups: normal group, model group and traditional Chinese medicine adjuvantgroup, high, medium and low dose group,15rats in each group, the self immunity for dailyintragastric administration to the termination of the experiment.(3) To observe the clinical signs: animal immune from the day two times a day toobserve the incidence of experimental animal, and neurological function score.The onsettime of0.5days for unit record rat onset (latency), and record the weight and the incidenceof nerve function, rats were.(4) sample collection:using retro-orbital venous plexus blood collection in the first14days after immunization (peak incidence) and21days (recovery) take blood, serum wasseparated for determination; perfusion of the heart and the brain, the spinal cord is placedfixed in4%paraformaldehyde, embedded in paraffin.(5)Detection of indicators: hematoxylin eosin (HE) brain, spinal cord pathologicalchange of staining method; ELISA method for the determination of serum interleukin-6(IL-6), transforming growth factor-β (TGF-β) conten.(6)statistical processing.Packageforstatistical analysis using SPSS19.0software. Results:(1)Rat incidence, latency, clinical score and body weight change: The blank groupand adjuvant group without the disease, Bushen Jiedu Tongluo Fang in the high, low dosegroup, the incidence of rats compared with the model group had statistical significance(P=0.00,<0.01); The average latency of rats of Bushen Jiedu Tongluo Fang in comparedwith model group were significantly prolonged (P<0.05); Bushen Jiedu Tongluo Fang high,medium dose group rats, alleviate the peak incidence in the average weight of phase wassignificantly higher than that in model group, low dose group of Chinese medicine; BushenJiedu Tongluo Fang in the incidence peak of incidence, dose group rats remission of nervefunction score was significantly lower than that in the model group.(2) Comparative observation onpathologicalstudy: The blank group, adjuvant group ratbrain and spinal cord without inflammatory lesions, the rats of group EAE peak visible brainand spinal cord parenchyma small vascular engorgement, perivascular infiltration ofinflammatory cells, forming a "cuff" shape. The peak incidence of Bushen Jiedu TongluoFang in high, low dose group, CNS in rats inflammatory lesion number compared with themodel group decreased significantly (P<0.05), high dose group, Chinese medicine than thelow dose group (P<0.05), there was no significant difference between the high, middle dosegroup (P>0.05); The incidence of remission of Bushen Jiedu Tongluo Fang high, mediumdose group rats CNS inflammatory lesion number compared with the model group decreasedsignificantly (P<0.05), no significant difference with those of low dosage Chinese medicinegroup and the model group (P>0.05), compared with the high dose group, low dose ofChinese medicine group mean number increased significantly (P <0.05traditional ChineseMedicine), high, middle dose group showed no significant difference (P>0.05).(3) The expression of serum IL-6, TGF-βCompared with the normal group, model group, low dose of Chinese medicine grouprats and alleviate the peak incidence of IL-6serum levels were significantly different (P <0.05); Compared with the peak incidence with the model group, the serum IL-6content inrats of every group was significantly reduced, significant difference (P <0.05), remissioncompared with the model group, except the low-dose group, the remaining rats diseaseremission expression level of IL-6decreased, with a significant difference (P <0.05); Theadjuvant group, model group and traditional Chinese medicine high, low dose group ratsserum IL-6expression levels in remission with peak decreased obviously, significantdifference (P <0.05). In the peak incidence, compared with the control group, the expression of the adjuvantgroup, model group rats serum TGF-obviously decreased, there was significant difference(P <0.05); Compared with the model group, except for the adjuvant group, significantlydifferent expression levels of TGF-β in the other groups (P <0.01); Traditional Chinesemedicine each dose group were compared with the model group, adjuvant group, the serumTGF-β content were increased, there was statistically significant difference (P <0.01); Theincidence of remission: middle dose group, the serum TGF-β content than those in themodel group increased significantly (P <0.01), there was no significant difference amongother groups (P>0.05).Conclusion:(1) Bushen Jiedu Tongluo Decoction can significantly reduce the incidence rate of acuteEAE rats, alleviate the clinical symptoms, shorten the course of disease.(2) Bushen Jiedu Tongluo Decoction can reduce EAE in the central nervous system ofrat inflammatory, protect the nerve function of rats.(3) part of the mechanism of Bushen Jiedu Tongluo Fang for the role of MS possiblythrough down-regulation of IL-6expression in rat serum, or (and) on expression of TGF-β,thereby inhibiting the formation of Th17cells, regulatory Th17cells differentiation, inhibitinflammation and autoimmune reaction.(4) Compared with the Bushen Jiedu Tongluo Fang three dose group, high dose groupand medium dose group may have more advantages. |
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