摘要
目的:探讨郭氏背三针干预对缺氧缺血性脑损伤(HIBD)新生大鼠的有效性及其可能的调控机制。方法:选取7日龄新生SD大鼠采用随机数字表法分为正常组、假手术组、模型组、背三针组、鼠神经组,每组10只。采用改良Rice法建立HIBD大鼠模型,背三针组予以背三针针刺干预7 d,鼠神经组注射鼠神经生长因子(10μg/kg),干预7 d。参照Zea Longa评分法评估大鼠神经功能缺损;行苏木精-伊红染色和尼氏染色观察大鼠海马组织病理改变;应用免疫组织化学染色(IHC)及实时荧光定量聚合酶链式反应(qPCR)检测大鼠海马组织凋亡调控因子(Bcl-2)、Bcl2相关X蛋白(Bax)、神经胶质纤维酸性蛋白(GFAP)、缺氧诱导因子1α(HIF-1α)的表达。结果:HIBD大鼠模型表现出不同程度的脑组织形态异常,表现为病变侧大脑肿胀、软化、萎缩、梗死、液化。相比于模型组,背三针组和鼠神经组缺损评分显著降低(P<0.05),脑组织形态异常发生率显著降低(P<0.05);背三针组和鼠神经组在神经功能缺损评分和脑组织形态异常发生率上无显著差异(P>0.05)。病理染色显示,正常组与假手术组大鼠海马区脑组织染色均匀,结构致密,神经细胞轮廓清晰;模型组大鼠海马区脑组织染色不均,结构明显紊乱,细胞间隙增宽、肿胀或空泡样改变,尼氏体减少且染色稍浅;与模型组比较,背三针组和鼠神经组大鼠海马组织正常尼氏体增多,细胞损伤程度较低,细胞轮廓大致正常。免疫组化结果显示,相比于正常组和假手术组,模型组大鼠海马组织Bax表达升高(P<0.05),Bcl-2表达降低(P<0.05);背三针组和鼠神经组Bax表达下调(P<0.05),Bcl-2上调(P<0.05),且背三针组Bax表达下调,Bcl-2上调幅度大于鼠神经组(P<0.05)。qPCR结果显示,相比于正常组和假手术组,模型组大鼠海马组织GFAP、HIF-1α表达显著增加(P<0.05);与模型组相比,背三针组和鼠神经组GFAP、HIF-1α的表达显著降低(P<0.05),且背三针组GFAP、HIF-1α的表达降低幅度显著于鼠神经组(P<0.05)。结论:背三针针刺干预可以改善HIBD大鼠的神经功能缺损和脑组织病理学损伤,其有效性可能是通过降低海马组织细胞凋亡,调控海马组织GFAP,HIF-1α和NGF的表达实现的。
Objective:To investigate the effect of Guo’s“Beisa zhen”acupuncture on neonatal rats with hypoxic-ischemic brain damage(HIBD)and its possible regulatory mechanism.Methods:Seven-day-old neonatal Sprague-Dawley rats were randomly divided into normal group,sham group,model group,acupuncture group and mouse nerve group(nerve growth factor group),with 10 rats in each group.Neurological deficit scores of rats was established by modified Rice method.Rats in acupuncture group was treated with Guo’s“Beisanzhen”acupuncture(once a day for 7 days),and mouse nerve growth factor(10μg/kg,once a day for 7 days)was injected to rats in mouse nerve group.The Zea Longa scoring criteria was used to assess the neurological deficits.Hematoxylin&eosin staining and Nissl’s staining were used to evaluate the pathological changes of hippocampal tissues of rats in different groups.Expressions of Bcl-2,Bax,GFAP and HIF-1αin hippocampal tissue were detected by immunohistochemical staining and quantitive real-time PCR.Results:HIBD rat model showed different degrees of alternations in cerebral histomorphology,including swelling,softening,atrophy,infarction and liquefaction on the lesion side.Compared to model group,the neurological deficit scores of rats in acupuncture group and mouse nerve group were significantly decreased,and the incidence of abnormal brain morphology was also significantly decreased notably,there was no significant difference in terms of neurological deficit score and the incidence of abnormal brain morphology between acupuncture group and mouse nerve group(P>0.05).Hematoxylin&eosin staining and Masson staining indicated uniform staining,dense structure,and clear contour of nerve cells of hippocampal tissue of rats in normal and sham groups.While hippocampal tissue of rats in model group was stained unevenly,the structure was obviously disordered,the intercellular space was widened,swollen or vacuolated,number of the Nissl body was reduced and the staining was slightly lighter.Compared to model group,pathological staining of hippocampal tissue of rats in acupuncture and mouse nerve groups showed increased number of Nissl body,reduced degree of cell damage and clear contour of nerve cells.Displayed through IHC,compared to normal and sham groups,expression of Bax was increased while expression of Bcl-2 was decreased in hippocampal tissues of rats in model group(P<0.05),expression of Bax in acupuncture group and mouse nerve group was decreased and Bcl-2 expression was increased(P<0.05),the effects of acupuncture group were more significantly than those in mouse nerve group(P<0.05).Displayed through qPCR,compared to normal and sham groups,expressions of GFAP,HIF-1αwere significantly increased in hippocampal tissues of rats in model group(P<0.05),the expressions of GFAP,HIF-1αin acupuncture group and mouse nerve group were reduced significantly(P<0.05),the changes of acupuncture group were more significant than those in mouse nerve group(P<0.05).Conclusion:Guo’s“Bei san zhen”acupuncture could improve the neurological deficit and cerebral histopathological injury of HIBD rats,which might be achieved by reducing the apoptosis of hippocampal tissue and regulating the expressions of GFAP,HIF-1αin hippocampal tissue.
作者
曹彩红
史华
单海军
介小素
侯玉晋
郭鑫
张英英
CAO Caihong;SHI Hua;SHAN Haijun;JIE Xiaosu;HOU Yujin;GUO Xin;ZHANG Yingying(He’nan University of Chinese Medicine,Zhengzhou,He’nan 450000)
出处
《中国中医药科技》
2025年第4期583-588,638,共7页
Chinese Journal of Traditional Medical Science and Technology
基金
河南中医药大学科研苗圃工程项目(MP2020-23)
河南省高等学校重点科研项目(22B360010)。
