摘要
[目的]研究黄精对中年房劳肾精亏虚模型大鼠免疫功能紊乱的调节作用。[方法]采用“中年雄鼠+雌雄同笼”方法制备中年房劳肾精亏虚大鼠模型,模拟中年男性因房劳致肾精亏虚引起的免疫功能紊乱状态。实验期间测定大鼠脾脏系数;苏木精-伊红(hematoxylin-eosin,HE)染色观察脾组织形态学变化;测定骨髓中性粒细胞和淋巴细胞占比,外周血淋巴细胞数量及其占比和中性粒细胞占比;脾淋巴细胞增殖试验检测脾脏淋巴细胞增殖能力;流式细胞术测定脾组织T淋巴细胞亚群占比;原位末端转移酶标记(terminal-deoxynucleoitidyl transferase mediated nick end labeling,TUNEL)检测脾组织细胞凋亡情况;免疫印迹检测脾组织与增殖相关蛋白磷酸化磷脂酰肌醇3-激酶(phospho-phosphatidylinositol 3-kinase,p-PI3K)、磷酸化蛋白激酶B(phospho-ptotein kinase B,p-AKT)和与凋亡相关蛋白B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X(Bcl-2 associated X,Bax)蛋白表达。[结结果]黄精可降低中年房劳肾精亏虚模型大鼠骨髓和外周血中性粒细胞占比,升高骨髓淋巴细胞占比,升高外周血淋巴细胞数量及其占比,降低脾组织T淋巴细胞(CD4^(+)/CD8^(+))比值,提高脾组织T淋巴细胞增殖能力,减少脾组织内的细胞凋亡,升高脾组织中与增殖相关蛋白p-PI3K和p-AKT的表达。[结论]黄精通过影响T淋巴细胞及其亚群占比,改善中年大鼠因肾精亏虚引起的免疫功能紊乱,其机制可能与调控脾组织与增殖相关蛋白PI3K/AKT的表达有关。
[Objective]To study the regulatory effect of Polygonatum sibiricum Red.on immune dysfunction in middle-aged rats with kidney essence deficiency caused by sexual overstrain.[Methods]To simulate the immune dysfunction caused by kidney essence deficiency due to sexual overstrain in middle-aged men,middle-aged rats with kidney essence deficiency model caused by sexual overstrain were prepared by“middle-aged male rats+male and female with the same cage”.The spleen coefficient of rats was measured during the experiment;the morphological changes of spleen were observed by hematoxylin-eosin(HE)staining;the proportion of neutrophils and lymphocytes in bone marrow was measured,and the number and proportion of lymphocytes in peripheral blood and the proportion of neutrophils were measured;splenic lymphocyte proliferation assay was used to detect splenic lymphocyte proliferation;the proportion of T lymphocyte subsets in spleen tissue was determined by flow cytometry;terminal deoxynucleoitidyl transferase mediated nick end labeling(TUNEL)was used to observe the apoptosis of spleen cells;Western blot was used to detect the protein expression of phospho-phosphatidylinositol 3-kinase(p-PI3K),phospho-protein kinase B(p-AKT),B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X(Bax)in spleen tissue.[Results]The results showed that Polygonatum sibiricum Red.could reduce the proportion of neutrophils in bone marrow and peripheral blood of middle-aged rats with kidney essence deficiency caused by sexual overstrain;increase the proportion of lymphocyte in bone marrow;increase the number and proportion of peripheral blood lymphocytes;reduce the ratio of T lymphocytes(CD4^(+)/CD8^(+))in spleen;improve the proliferation of T lymphocyte in spleen;reduce cell apoptosis in spleen tissue;increased spleen tissue proliferation related protein p-PI3K and p-AKT expression.[Conclusion]Polygonatum sibiricum Red.can improve the immune dysfunction caused by kidney essence deficiency due to sexual overstrain in middle-aged rats by affecting the proporti
作者
俞静静
杜宇忠
苏洁
颜美秋
胡雪灵
潘斌辉
牛壮伟
陈素红
吕圭源
YU Jingjing;DU Yuzhong;SU Jie(Zhejiang Chinese Medical University,Hangzhou,310053,China;Shanxi Medical University)
出处
《浙江中医药大学学报》
CAS
2024年第8期895-904,共10页
Journal of Zhejiang Chinese Medical University
基金
国家自然科学基金项目(82274139)
浙江省重点研发计划项目(2020C04020)。
关键词
黄精
中年房劳肾精亏虚模型
免疫功能紊乱
T淋巴细胞
增殖
凋亡
PI3K/AKT
Polygonatum sibiricum Red.
middle-aged model with kidney essence deficiency caused by sexual overstrain
immune dysfunction
T lymphocyte
proliferation
apoptosis
PI3K/AKT