摘要
目的比较分别含红芪、黄芪的益气养血汤和补中益气汤含药血清对SAMP8小鼠脾淋巴细胞免疫功能影响差别。方法 50只青龄昆明种小鼠随机分为益气养血汤红芪组、益气养血汤黄芪组、补中益气汤红芪组、补中益气汤黄芪组、空白组,每组10只,连续灌胃14天,制备含药血清和空白血清。提取SAMP8小鼠脾淋巴细胞,分为益气养血汤红芪含药血清组、益气养血汤黄芪含药血清组、补中益气汤红芪含药血清组、补中益气汤黄芪含药血清组和空白血清组,以相应含药血清和空白血清培养;SAMP8鼠组和青龄鼠组以完全培养液分别培养SAMP8小鼠和青龄小鼠脾淋巴细胞。不同质量分数(20%、40%、80%)含药血清和空白血清培养SAMP8小鼠脾淋巴细24、48、72 h,MTT法检测各组脾淋巴细胞增殖功能,确定最佳含药血清质量分数和作用时间。另取SAMP8和青龄小鼠脾淋巴细胞,4个含药血清组和空白血清组以最佳质量分数血清培养SAMP8小鼠脾淋巴细胞至最佳作用时间,SAMP8鼠组和青龄鼠组以完全培养液分别培养SAMP8小鼠和青龄小鼠脾淋巴细胞48 h,流式细胞仪检测脾T淋巴细胞表面CD28^+、CD152^+分子的表达,ELISA法检测培养上清液中TNF-α的表达,实时荧光定量PCR和Western Blot检测脾T淋巴细胞PI3K mRNA和蛋白的表达。结果 MTT结果显示,当分别含红芪和黄芪的益气养血汤含药血清质量分数为40%培养48 h,分别含红芪和黄芪的补中益气汤含药血清质量分数为40%培养72 h时,SAMP8小鼠脾淋巴细胞增殖效果最佳。与青龄鼠组比较,SAMP8鼠组小鼠脾T淋巴细胞表面CD28^+分子表达下降,CD152^+分子表达增高(P<0.05),TNF-α表达及PI3K mRNA和蛋白表达降低(P<0.05)。与空白血清组比较,分别含红芪和黄芪的益气养血汤和补中益气汤含药血清组小鼠脾T淋巴细胞表面CD28^+分子表达升高,CD152^+分子表达降低(P<0.05),TNF-α表达及PI3K mRNA和蛋白表达�
Objective To compare the different effects between Yiqi Yangxue and Buzhong Yiqi Decoction Containing Serums with Radix Hedysari or Radix Astragali on immunologic function of SAMP8 mice splenic lymphocytes. Methods Fifty young Kunming mice were randomly divided into 5 groups:Yiqi Yangxue Decoction with Radix Hedysari(YYDRH) group, Yiqi Yangxue Decoction with Radix Astragali(YYDRA) group, Buzhong Yiqi Decoction with Radix Hedysari(BYDRH) group, Buzhong Yiqi Decoction with Radix Astragali(BYDRA) group and blank group, 10 in each group. Normal saline and medicine were continuous intragastric administrated for 14 days to prepare drug-containing serums and blank serum. Splenic lymphocytes were divided into 5 groups: YYDRH containing serum group, YYDRA containing serum group, BYDRH containing serum group, BYDRA containing serum group and blank serum group, and were cultured with serums accordingly. The splenic lymphocytes of SAMP8 mice group and young mice group were cultivated with complete medium. To determine the optimal serum concentration and culture time on promoting proliferation of SAMP8 mice splenic lymphocytes, MTT assay was performed after culturing 24 h, 48 h and 72 h with 20%, 40%, 80% concentration of medicated serums and blank serum respectively. SAMP8 mice splenic lymphocytes were exposed with the best concentration of medicated serums and blank serum to optimal culture time. SAMP8 mice group and young mice group were cultured with complete medium for 48 h. Expressions of CD28^+ and CD152^+ molecules on the surface of splenic T lymphocytes were measured by flow cytometry. The level of TNF-α in lymphocytes culture supernatant was tested by ELISA. Real time fluorescence quantitative PCR and Western blot were used to detect expression of PI3K mRNA and its protein. Results MTT results showed that YYDRH/YYDRA and BYDRH/BYDRA containing serums could enhance the proliferation of SAMP8 mice splenic lymphocytes, and the effect was time and dosage dependent. The best efficacy in enhancing lymphocytes proliferati
作者
杨思霞
李纬
孙美花
张文君
张高林
程卫东
YANG Si-xia;LJ Wei;SUN Mei-hua;ZHANG Wen-jun;ZHANG Gao-lin;CHENG Wei-dong(College of Traditional Chinese Medicine,Southern Medical University,Guangzhou,510515;Oncology Department of Gansu Provincial Hospital of Traditional Chinese Medicine,Lanzhou,730050;Department of Traditional Chinese Medicine,Gansu traditional Chinese Medicine School,Lanzhou ,730050)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2019年第3期335-341,共7页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金面上项目(No.81373806)
