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目的:通过研究比较黄芪种子与饮片对环磷酰胺致免疫功能低下模型大鼠相关凋亡基因Fas/Fas L表达的作用差异,探讨、评价黄芪种子作为药用新资源的开发利用前景。方法:将SD雄性大鼠随机分为正常对照组(NC)、模型组(M)、黄芪饮片组(HYG)、黄芪种子组(HZG)4组,每组10只,除NC组外其余各组采用环磷酰胺法(Cy,80 mg·kg~(-1)·d~(-1))复制免疫功能低下模型大鼠。造模成功后,HYG和HZG组灌胃给予黄芪饮片、种子水煎剂(20 g·kg~(-1)·d~(-1)),其余组给予等量的生理盐水,每日1次,连续7 d。7 d后,处死动物,取血清、肝脏等备用,ELISA法检测血清IgA、IgM、IgG含量及CD3+、CD_4~+、CD_8~+比值;Rt-PCR检测Fas/Fas L基因的表达;采用HE染色观察肝脏组织病理变化。结果:M组动物肝小叶及血窦呈高度扩张瘀血,细胞呈水样变性,黄芪等治疗组肝脏组织病理损伤较M组明显改善。M组动物血清IgA、IgM、IgG的含量明显高于NC组(P<0.01);HYG组血清IgA、IgM、IgG的含量明显低于M组(P<0.05);与HYG组比较,HZG组血清IgA、IgM、IgG的含量明显降低(P<0.01)。与NC比较,M组血清CD_4~+/CD_8~+比值明显降低(P<0.01);HZG及HYG较M组血清CD_4~+/CD_8~+含量明显增加(P<0.01)。与NC组比较,M组的Fas、Fas L m RNA的表达量显著降低(P<0.01);HZG和HYG组Fas、Fas L m RNA的表达量明显低于M组(P<0.01);与HZG比较,HYZ组Fas、Fas L m RNA的表达量更低(P<0.05)。结论:黄芪种子及饮片组能明显降低Cy模型动物血清IgA、IgM、IgG的含量,增加CD_4~+/CD_8~+比值,下调肝组织凋亡基因Fas、Fas L m RNA的表达,对抗Cy所致的免疫抑制性肝脏损伤,提示黄芪种子可作为黄芪药材新的入药部位,具有极大的开发潜力和良好的开发前景。
OBJECTIVE: To study the differences of the expression of Fas / Fas Ligand and Fas / Fas Ligand in the immunosuppressive model induced by cyclophosphamide in Astragalus mongholicus to explore and evaluate the development and utilization of Astragalus spp. As a new medicinal resource. Methods: SD male rats were randomly divided into normal control group (NC), model group (M), astragalus extract group (HYG), Astragalus seed group (HZG) 4 groups, Groups were immunosuppressed with Cyclophosphamide (Cy, 80 mg · kg -1 · d -1). After the model was established successfully, the HYG and HZG groups were given gavage with Astragalus mongholicus and seeds decoction (20 g · kg -1 · d -1), while the rest group were given the same amount of saline daily Times, for 7 days. After 7 days, the animals were sacrificed and the serum and liver were taken for spare. Serum IgA, IgM and IgG levels and the ratio of CD3 +, CD4 + and CD8 + were detected by ELISA. The expression of Fas / Fas L gene was detected by Rt- Staining to observe the pathological changes of liver tissue. Results: The hepatic lobules and sinusoids in group M were highly stasis, and the cells were degenerated in water. The pathological changes of liver tissues in astragalus mongholicus group were significantly improved compared with those in M group. The levels of serum IgA, IgM and IgG in M group were significantly higher than those in NC group (P <0.01). The levels of serum IgA, IgM and IgG in HYG group were significantly lower than those in M group (P <0.05). Compared with HYG group, Serum IgA, IgM, IgG levels were significantly lower (P <0.01). Compared with NC, the serum CD_4 ~ + / CD_8 ~ + ratio in M group was significantly lower than that in NC group (P <0.01). The levels of CD_4 ~ + / CD_8 + in serum of HZG and HYG groups were significantly increased (P <0.01) Compared with NC group, the expression of Fas and Fas LmRNA in M group was significantly lower than that in M group (P <0.01); the expression of Fas and Fas LmRNA in HZG and HYG groups was significantly lower than that in M group (P <0.01); Compared with HZG group, the expression of Fas and Fas LmRNA in HYZ group was lower (P <0.05). Conclusion: The Astragalus mongholicus seeds and Decoction group can significantly reduce the levels of serum IgA, IgM and IgG, increase the ratio of CD_4 ~ + / CD_8 ~ + and downregulate the expression of Fas and Fas LmRNA in liver tissue of Cy model rats, Induced immunosuppressive liver injury, suggesting that astragalus seed astragalus medicine can be used as a new drug site, has great potential for development and good development prospects.