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目的观察人参三七川芎提取物对自然衰老大鼠胸主动脉及骨架蛋白的影响。方法 50只SD雄性大鼠(8~9月龄)按体重随机分为模型组、中药低、中、高剂量组及白藜芦醇组,每组各10只,模型组大鼠正常饲养至21月龄;各给药组大鼠分别从18月龄开始药物干预,中药低、中、高剂量组给药剂量分别为576、1 152、2 305 mg/kg,白藜芦醇组给药剂量为50 mg/kg,模型组按体重给予等体积的纯净水,每日1次,连续灌胃3个月;同时以10只SD雄性大鼠(3月龄)作为青年对照组(青年组)。应用HE染色观察胸主动脉血管形态的改变,β-半乳糖苷酶(SA-β-gal)特异染色法计算蓝染组织比率,免疫荧光染色观察骨架蛋白F-actin形态的改变,Western blot法检测骨架蛋白F-actin和SM22α表达情况。结果与青年组比较,模型组血管内膜损伤且不光滑,中膜明显增生且排列紊乱,弹力板扭曲、断裂、结构不完整,外膜增生明显,SA-β-gal染色组织蓝染比率明显增加(P<0.01),F-actin荧光强度及蛋白表达明显增多(P<0.01),而SM22α蛋白表达明显减少(P<0.01)。药物干预后,各给药组血管壁内皮轻度受损,弹力膜结构较完整,中膜及外膜厚度明显降低(P<0.01);SA-β-gal染色中药中剂量及白藜芦醇组蓝染比率减少(P<0.05),而中药低、高剂量蓝染比率改变无统计学意义(P>0.05);各给药组F-actin荧光强度明显降低(P<0.01);蛋白表达方面,中药中、高剂量组及白藜芦醇组F-actin蛋白表达减少(P<0.01),而中药低剂量组改变则无统计学意义(P>0.05),各给药组SM22α蛋白表达增加(P<0.01)。结论细胞骨架微丝蛋白F-actin形态及蛋白表达在衰老过程中改变明显,其相关蛋白SM22α蛋白表达亦改变明显,可能共同参与了血管老化的进程;人参三七川芎提取物可以一定程度上延缓血管老化,且对于血管F-actin和SM22α蛋白有明显干预作用,可能间接通过此作用延缓了血管老化的进程。
Objective To observe the effect of extract of ginseng, rhizoma chuanxiong and rhizoma chuanxiong rhizome on thoracic aorta and skeletal protein in rats with natural aging. Methods Fifty male Sprague-Dawley rats (8-9 months old) were randomly divided into model group, low dose group, medium dose group and high dose resveratrol group and 10 rats in each group. The rats in model group were fed to the normal 21 months old. The rats in each treatment group started drug intervention from 18 months of age respectively. The doses of low, medium and high doses of Chinese herbs were 576,1 152 and 2 305 mg / kg, respectively. Resveratrol A dose of 50 mg / kg, the model group according to body weight given an equal volume of purified water, once a day, continuous gavage for 3 months; 10 SD male rats (3 months old) as a youth control group (youth group ). The morphology of the thoracic aorta was observed by HE staining. The ratio of blue staining was calculated by β-galactosidase (SA-β-gal) staining. The morphological changes of F-actin were observed by immunofluorescence staining. Detection of skeletal protein F-actin and SM22α expression. Results Compared with the young group, the intima of the model group was not smooth and smooth, the media was hyperplastic and disordered, the twisting and rupture of the elastic plate, the incomplete structure and the adventitial hyperplasia were obvious. The ratio of blue dye in the SA-β-gal staining tissue was obvious (P <0.01), the fluorescence intensity and protein expression of F-actin increased significantly (P <0.01), while the expression of SM22α protein decreased significantly (P <0.01). After drug intervention, the endothelium of the vessel wall in each administration group was mildly damaged, the structure of the elastic membrane was relatively intact, and the thickness of the media and adventitia was significantly decreased (P <0.01). The middle dose of SA-β-gal staining and resveratrol (P <0.05), but there was no significant difference in the blue-dye ratio between low and high doses of traditional Chinese medicine (P> 0.05). The fluorescence intensity of F-actin in each group was significantly decreased (P <0.01) (P <0.01), while there was no significant difference in the low-dose group (P> 0.05). The expression of SM22α in each group was significantly lower than that in the middle-dose and high-dose groups Increase (P <0.01). CONCLUSION: The morphology and protein expression of F-actin in cytoskeletal skeletal fibroblasts changed significantly during aging, and the expression of related protein SM22α also changed obviously, which may participate in the process of vascular aging. The extract of Panax Chuanxiong could delay to a certain degree Vascular aging, and for the vascular F-actin and SM22α protein significant intervention, may indirectly through this delay in the process of vascular aging.