基于非稳态负荷评估雄性SD大鼠衰老进展及左归丸的干预作用  

Using allostatic load as a tool for evaluating aging of male SD rats and the intervention effects of Zuogui Pill

作　　者：贾雨欣 邓晓红 孙磊峰 陈丹丹 孙贤俊 段晓红 严诗楷[2] 黄建华 JIA Yuxin;DENG Xiaohong;SUN Leifeng;CHEN Dandan;SUN Xianjun;DUAN Xiaohong;YAN Shikai;HUANG Jianhua(Institute of Integrated Traditional Chinese and Western Medicine,Huashan Hospital Affiliated to Fudan University,Shanghai 200040,China;School of Pharmacy,Shanghai Jiao Tong University,Shanghai 200240,China)

机构地区：[1]复旦大学附属华山医院中西医结合研究所,上海200040 [2]上海交通大学药学院,上海200240

出　　处：《中国实验动物学报》2024年第6期753-761,共9页Acta Laboratorium Animalis Scientia Sinica

基　　金：上海市扬帆计划(21YF1404000)。

摘　　要：目的采用非稳态负荷(allostatic load,AL)的理论与方法评估衰老以及左归丸对自然衰老雄性SD大鼠模型的影响。方法自然衰老雄性SD大鼠,分别在2、5、8、14、18、21月龄剪尾取血,其中18月龄时随机分为老年对照组、左归丸低剂量组和左归丸高剂量组。左归丸干预3个月后,取用血样以及肌肉、肝等组织样本。血样用流式细胞术检测T淋巴细胞数量和凋亡率,并检测血清生化指标,包括低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-c)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-c)、甘油三酯(triglycerides,TG)、总胆固醇(total cholesterol,TC)、游离脂肪酸(free fatty acids,FFA)、25羟维生素D(25 hydroxyvitamin D,25-OH-D)、皮质酮(corticosterone,CORT)、C反应蛋白(C-reactive protein,CRP)、白细胞介素-6(interleukin 6,IL-6)。将LDL-c、TC、HDL-c、FFA、TG、CORT、IL-6、CRP、25-OH-D、CD3^(+)T淋巴细胞计数、CD3^(+)T淋巴细胞凋亡率共11项指标用于AL评分。结果大鼠血清LDL-c、TG、TC、25-OH-D、CRP、IL-6随增龄发生显著改变,但变化模式有所不同。CD3^(+)T淋巴细胞计数随增龄显著降低(P<0.01),CD3^(+)、CD4^(+)、CD8^(+)T淋巴细胞凋亡率随增龄显著升高(P<0.01)。左归丸干预后,老年大鼠血清CORT水平显著提高(P<0.01)、CD8^(+)T淋巴细胞凋亡率显著降低(P<0.05)。AL评分自5月龄开始升高,至18月龄到达峰值。结论与单一指标比较,AL能更好地表征衰老过程。左归丸可以提高衰老大鼠的应激反应能力,延缓其免疫衰老。Objective Using the concept of allostatic load(AL)to evaluate aging of male SD rats and the effectiveness of Zuogui Pill in naturally aging rats.Methods Naturally aging male SD rats were tested at the ages of 2,5,8,14,18,and 21 months.They were divided into an elderly control group,low⁃dose Zuogui Pill group,and high⁃dose Zuogui Pill group.Intervention with Zuogui Pill was trialed for 3 months.Blood samples were taken from the tails of rats each month,and the number of T lymphocytes and rate of apoptosis were measured by flow cytometry.Low⁃density lipoprotein cholesterol(LDL⁃c),high⁃density lipoprotein cholesterol(HDL⁃c),triglycerides(TG),total cholesterol(TC),free fatty acids(FFA),25 hydroxyvitamin D(25⁃OH⁃D),corticosterone(CORT),C⁃reactive protein(CRP),and interleukin⁃6(IL⁃6)were detected in rat sera.By identifying the collinearity between indicators and professional considerations,LDL⁃c,TC,HDL⁃c,FFA,TG,CORT,IL⁃6,CRP,25⁃OH⁃D,CD3^(+)T cell count,and CD3^(+)T cell apoptosis rate were included in the AL scoring.The threshold for each indicator was established with data from 5⁃month⁃old rats,and the score was 1 point below or/and above the threshold.Results The serum levels of LDL⁃c,TG,TC,25⁃OH⁃D,CRP,and IL⁃6 of rats showed significant changes with age,although the patterns of change differed.The CD3^(+)T lymphocyte count significantly decreased with age(P<0.01),while the apoptosis rates of CD3^(+),CD4^(+),and CD8^(+)T lymphocytes significantly increased with age(P<0.01).Zuogui Pill significantly increased serum CORT levels in elderly rats(P<0.01)and reduced the apoptosis rate of CD8^(+)T lymphocytes(P<0.05).The AL score began to increase in rats at 5 months of age and reached its peak in those of 18 months of age.Conclusions AL can better characterize the aging process compared to a single indicator.Zuogui Pill can improve the stress response ability of aging rats and alleviate immunosenescence.

关 键 词：非稳态负荷 衰老 左归丸 免疫衰老 

分 类 号：Q95-33[生物学—动物学]