Shenzhen Key Laboratory of Hospital Chinese Medicine Preparation, Shenzhen Traditional Chinese Medicine Hospital, Guangzhou University of Chinese Medicine, Shenzhen, Guangdong, China.
Division of Life Science and Center for Chinese Medicine, The Hong Kong University of Science and Technology, Hong Kong, China.
BMC Complement Altern Med. 2018 Apr 3;18(1):119. doi: 10.1186/s12906-018-2146-4.
A Chinese herbal formula, namely Jian-Pi-Yi-Shen (JPYS), has been clinically prescribed for patients with chronic kidney disease associated-anemia, and which can improve the patient's immunological system. However, the mechanisms of JPYS involved in anemia and immune response have not been investigated. To study the role of JPYS in regulating hematopoietic and immunological functions, we investigated its activities on the expressions of erythropoietin and pro-inflammatory cytokines in cultured cells.
The standardized herbal extracts of JPYS (0-30 μg/ml) were applied onto cultured cells for 24-48 h. Total RNA was collected from the treated cells and subjected to real-time quantitative PCR analysis. Cultured HEK293T cells, transfected with a construct composed of hypoxia response element tagged with a luciferase gene, i.e. pHRE-Luc, were treated with JPYS extracts (1-30 μg/ml) for 24 h. The cell lysates were subjected to luciferase assay.
The treatment with JPYS extract onto cultured HEK293T cells induced erythropoietin expression in a dose-dependent manner, having the highest response by ~ 50% of increase. In parallel, application of JPYS extract for 24 h stimulated expressions of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α in cultured RAW 264.7 macrophages. In contrast, the pretreatment with JPYS extract suppressed expressions of IL-1β, IL-6, and TNF-α in lipopolysaccharide-induced macrophages.
These results confirmed the hematopoietic function of JPYS in regulating erythropoietin expression, as well as the bidirectional immune-modulatory roles of JPYS by regulating the expression of pro-inflammatory cytokines in cultures.
一种名为健脾益肾（JPYS）的中草药配方已在临床上用于治疗伴有贫血的慢性肾脏病患者，可改善患者的免疫系统。然而，JPYS 改善贫血和免疫反应的机制尚未得到研究。为了研究 JPYS 在调节造血和免疫功能中的作用，我们研究了它对培养细胞中促红细胞生成素和促炎细胞因子表达的影响。
将 JPYS 的标准化草药提取物（0-30μg/ml）应用于培养细胞 24-48 小时。从处理过的细胞中收集总 RNA，并进行实时定量 PCR 分析。用 JPYS 提取物（1-30μg/ml）处理培养的 HEK293T 细胞 24 小时，转染由缺氧反应元件标记的荧光素酶基因构建体，即 pHRE-Luc。将细胞裂解物用于荧光素酶测定。
JPYS 提取物处理培养的 HEK293T 细胞可诱导促红细胞生成素表达呈剂量依赖性，最高反应增加约 50%。同时，JPYS 提取物应用 24 小时刺激培养的 RAW 264.7 巨噬细胞中白细胞介素（IL）-1β、IL-6 和肿瘤坏死因子（TNF）-α的表达。相反，JPYS 提取物预处理可抑制脂多糖诱导的巨噬细胞中 IL-1β、IL-6 和 TNF-α的表达。
这些结果证实了 JPYS 的造血功能可通过调节促红细胞生成素的表达来调节贫血，以及 JPYS 通过调节培养物中促炎细胞因子的表达来发挥双向免疫调节作用。
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