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目的探讨益肾活血法对实验性变态反应性脑脊髓炎(EAE)大鼠模型的影响及对神经干细胞增殖分化的机制。方法构建EAE大鼠模型,将大鼠分为正常对照组、EAE对照组和低、中、高剂量益肾活血法组,根据病理学方法检测大鼠发病时间和高峰期神经功能障碍评分,ELISA检测IL-17、IL-4、IFL-γ浓度,流式细胞仪检测CD4+和CD8+比例,Brd U标记法检测神经干细胞增殖,RT-PCR检测Hes1和Mash1 mRNA表达。结果与EAE对照组比较,低、中、高剂量组潜伏期显著增加,进展期和神经功能障碍评分显著降低(P<0.01);EAE对照组和低、中、高剂量组脑组织匀浆中IFL-γ和IL-17浓度显著高于正常对照组,IL-4浓度显著低于正常对照组;低、中、高剂量组IFL-γ和IL-17浓度显著低于EAE对照组,IL-4浓度显著高于EAE对照组(P<0.01);与正常对照组比较,EAE对照组和低、中、高剂量组血中CD4+/CD8+显著降低(P<0.01),与EAE对照组比较显著上升(P<0.01);与EAE对照组比较,中剂量组Brd U阳性细胞平均光密度显著高于低剂量组,高剂量组高于中剂量组(P<0.01);与EAE对照组比较,低、中、高剂量组均能提高Hes1和Mash1 mRNA表达量。结论益肾活血法能延长EAE大鼠发病潜伏期,缩短进展期,降低发病高峰期神经功能受损;升高CD4+和CD8+比例;降低促炎因子IFL-γ和IL-17分泌,升高抗炎因子IL-4的分泌;通过上调Hes1增加神经干细胞增殖。
Objective To investigate the effect of Yishenhuoxue Therapy on experimental allergic encephalomyelitis (EAE) rat model and its mechanism of proliferation and differentiation of neural stem cells. Methods EAE rat model was established. The rats were divided into normal control group, EAE control group and low, middle and high dose of Yishenhuoxue method group, according to the pathological method, the onset time and peak neurological deficit score The concentrations of IL-17, IL-4 and IFL-γ were detected by flow cytometry. The proportion of CD4 + and CD8 + was detected by flow cytometry. The proliferation of neural stem cells was detected by BrdU labeling. The expression of Hes1 and Mash1 mRNA was detected by RT-PCR. Results Compared with EAE control group, the incubation period in low, medium and high dose groups significantly increased, and the score of progressive and neurological dysfunction decreased significantly (P <0.01). In EAE control group and low, medium and high dose groups, the contents of IFL The concentrations of IL-17 and IL-17 in serum were significantly higher than those in normal control group (P <0.05). The concentrations of IL-4 and IL-4 in serum were significantly lower than those in normal control group (P <0.01). Compared with the normal control group, the levels of CD4 + / CD8 + in the EAE control group and the low, medium and high dose groups were significantly decreased (P <0.01) and significantly increased compared with the EAE control group (P <0.01). Compared with EAE control group, the average optical density of BrdU positive cells in middle dose group was significantly higher than that in low dose group and high dose group was higher than that in middle dose group (P <0.01). Compared with EAE control group, , Medium and high dose group can increase the expression of Hes1 and Mash1 mRNA. Conclusion Yishenhuoxue can prolong the incubation period, shorten the progression of EAE rats, reduce the impaired neurological function during peak incidence, increase the proportion of CD4 + and CD8 +, decrease the secretion of proinflammatory cytokines (IFL-γ and IL-17), increase the anti-inflammatory cytokines IL -4 secretion; increase neural stem cell proliferation by up-regulating Hes1.