[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.
Study on the Effect of Polygonatum Sibiricum Red.on Immune Dysfunction of Middle-aged Rats with Kidney Essence Defi-ciency
[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 proportion of T lymphocytes and their subpopulations,and its mechanism may be related to regulating the spleen tissue proliferation related protein expression of PI3K/AKT.
Polygonatum sibiricum Red.middle-aged model with kidney essence deficiency caused by sexual overstrainimmune dysfunctionT lymphocyteproliferationapoptosisPI3K/AKT