Liu Zhi-Jia, Wang Cun-Ping, Yang Bin, Wang Nan, Wang Qin, Gu Jian, Tan Rui
College of Pharmacy, Southwest Minzu University Chengdu 610041, China.
College of Life Science and Engineering, Southwest Jiaotong University Chengdu 610031, China.
Zhongguo Zhong Yao Za Zhi. 2022 Apr;47(8):2064-2073. doi: 10.19540/j.cnki.cjcmm.20211216.705.
To study the protective effect of Ershiwuwei Zhenzhu Pills on ischemic stroke rats. Ninety 4-weeks-old SPF male SD rats were randomly divided into 6 groups(n=15):sham operation group, model group, nimodipine group(12 mg·kg(-1)), Ershiwuwei Zhenzhu Pills high-dose group(400 mg·kg(-1)), Ershiwuwei Zhenzhu Pills medium-dose group(200 mg·kg(-1)), Ershiwuwei Zhenzhu Pills low-dose group(100 mg·kg(-1)).The permanent middle cerebral artery occlusion model(PMCAO) was established in the model group, nimodipine group, and Ershiwuwei Zhenzhu Pills groups by the improved thread plug method, while the sham operation group did not insert the thread plug.Nimodipine group and Ershiwuwei Zhenzhu Pills groups were given intragastric administration once a day for 24 days before the modeling operation, and once 1 hour before the modeling operation, while sham operation group and model group were given equal volumes of distilled water.The neuroethology of the surviving rats was measured; The volume of cerebral infarction in rats was measured by TTC method; The histopathology of rat brain was observed by HE method; The expression levels of tumor necrosis factor α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),malondialdehyde(MDA),superoxide dismutase(SOD) and catalase(CAT) in serum were detected by ELISA;The mRNA expressions of Notch 1,Jagged 1,Hes 1 and Bcl-2 in rat brain were detected by RT-PCR;Western blot was used to detect the expression levels of caspase-3 protein in rat brain; the expression levels of vascular endothelial growth factor(VEGF) and CD34 positive cells in rat brain were detected by immunofluorescence.The low, medium and high dose groups of Ershiwuwei Zhenzhu Pills and nimodipine group could significantly reduce the neurobehavioral score and cerebral infarction volume of rats with permanent middle cerebral artery occlusion, reduce the morphological changes of nerve cells, decrease the expression of TNF-α,IL-1β and IL-6 in rat serum, increase the activity of SOD and CAT,and reduce the level of MDA.Furthermore, the expression levels of Notch l, Jagged l, Hes l and Bcl-2 mRNA were significantly increased, and the expression level of caspase-3 protein was decreased.Meanwhile, the number of VEGF and CD34 positive cells increased in the treatment group.The differences were statistically significant. Ershiwuwei Zhenzhu Pills has a protective effect on ischemic stroke rats, and its mechanism may be related to anti-inflammation, anti-oxidation, promotion of nerve cell proliferation, inhibition nerve cell apoptosis and promotion of angiogenesis.
探讨二十五味珍珠丸对缺血性脑卒中大鼠的保护作用。将90只4周龄的SPF级雄性SD大鼠随机分为6组(n = 15):假手术组、模型组、尼莫地平组(12 mg·kg⁻¹)、二十五味珍珠丸高剂量组(400 mg·kg⁻¹)、二十五味珍珠丸中剂量组(200 mg·kg⁻¹)、二十五味珍珠丸低剂量组(100 mg·kg⁻¹)。采用改良线栓法对模型组、尼莫地平组及二十五味珍珠丸各剂量组建立永久性大脑中动脉闭塞模型(PMCAO),假手术组不插入线栓。尼莫地平组和二十五味珍珠丸各剂量组在造模手术前24天每天灌胃给药1次,造模手术前1小时再给药1次,假手术组和模型组给予等体积蒸馏水。测定存活大鼠的神经行为学;采用TTC法测定大鼠脑梗死体积;采用HE法观察大鼠脑组织病理学;采用ELISA法检测血清中肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、丙二醛(MDA)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的表达水平;采用RT-PCR法检测大鼠脑组织中Notch 1、Jagged 1、Hes 1和Bcl-2的mRNA表达;采用Western blot法检测大鼠脑组织中caspase-3蛋白的表达水平;采用免疫荧光法检测大鼠脑组织中血管内皮生长因子(VEGF)和CD34阳性细胞的表达水平。二十五味珍珠丸低、中、高剂量组及尼莫地平组均可显著降低永久性大脑中动脉闭塞大鼠的神经行为学评分和脑梗死体积,减轻神经细胞形态学改变,降低大鼠血清中TNF-α、IL-1β和IL-6的表达,提高SOD和CAT活性,降低MDA水平。此外,Notch 1、Jagged 1、Hes 1和Bcl-2 mRNA表达水平显著升高,caspase-3蛋白表达水平降低。同时,治疗组VEGF和CD34阳性细胞数量增加。差异具有统计学意义。二十五味珍珠丸对缺血性脑卒中大鼠具有保护作用,其机制可能与抗炎、抗氧化、促进神经细胞增殖、抑制神经细胞凋亡及促进血管生成有关。