Hou Jianhao, Qian Jinjun, Li Zhenlin, Gong Aixiu, Zhong Sixia, Qiao Li, Qian Shihui, Zhang Yanxin, Dou Renjie, Li Rui, Yang Ye, Gu Chunyan
The Third Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210001, People's Republic of China.
School of Medicine and Holistic Integrative Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, People's Republic of China.
Onco Targets Ther. 2020 Jan 31;13:959-973. doi: 10.2147/OTT.S235944. eCollection 2020.
(L.) Medik. (Malvaceae) derived Huangkui capsules (HKC) represent a traditional Chinese medicine that has been widely applied to the clinical therapy of kidney and inflammatory diseases. The present study aimed to determine the potential therapeutic effects and underlying mechanisms of the ingredients on Multiple Myeloma (MM), an incurable disease that exhibits malignant plasma cell clonal expansion in the bone marrow.
A 5TMM3VT syngeneic MM-prone model was established and treated with HKC. Murine pre-osteoblast MC3T3-E1 and pre-osteoclast Raw264.7 cells were treated with nine flavonoid compounds extracted from the flowers of . MC3T3-E1 and Raw264.7 cells were then examined by alizarin red staining and tartrate-resistant acid phosphatase activity staining, respectively. The proliferation of two human MM cells (ARP1, H929) was examined by performing an MTT assay following treatment with flavonoid compounds. Additionally, the cell cycle was analyzed via staining and flow cytometry. The differential expressions of certain proteins were detected via Western blotting, transcriptomic RNA-sequencing as well as RT-qPCR.
The results revealed that MM-prone animals appeared to be protected following HKC treatment, as evidenced by a prolonged survival rate. Furthermore, four of the nine flavonoid compounds [Hyperin/Hyperoside, HK-2; Cannabiscitrin, HK-3; 3--kaempferol-3--acetyl-6--(p-coumaroyl)-β-D-glucopyranoside, HK-11; 8-(2''-pyrrolidione-5''-yl)-quercetin, HK-B10] induced the differentiation of murine pre-osteoblast MC3T3-E1 cells. In addition, two compounds [Isomyricitrin, HK-8; quercetin-8-(2''-pyrrolidione-5"-yl)-3'--β-D-glucopyranosid, HK-E3] suppressed osteoclastogenesis in murine Raw264.7 cells. HK-11 directly inhibited MM cells (ARP1 and H929) proliferation and induced G0/G1 cell cycle arrest, which may have involved the suppressing β-catenin protein, increasing expressions of IL-6 and TNF-α, as well as activating mature TGF-β1 and some other metabolic pathways.
These results of the present study indicated that the bio-active ingredients of HKC exerted protective effects on MM mouse survival through promoting osteoblastogenesis and suppressing osteoclastogenesis, thus improving the bone marrow microenvironment to inhibit MM cell proliferation.
锦葵科植物黄葵（Abelmoschus moschatus (L.) Medik.）衍生的黄葵胶囊（HKC）是一种传统中药，已广泛应用于肾脏疾病和炎症性疾病的临床治疗。本研究旨在确定其成分对多发性骨髓瘤（MM）的潜在治疗作用及潜在机制，MM是一种无法治愈的疾病，其特征为骨髓中恶性浆细胞克隆性扩增。
建立5TMM3VT同基因MM易感模型并用HKC进行治疗。用从黄葵花中提取的9种黄酮类化合物处理小鼠前成骨细胞MC3T3-E1和前破骨细胞Raw264.7细胞。然后分别通过茜素红染色和抗酒石酸酸性磷酸酶活性染色检测MC3T3-E1和Raw264.7细胞。用黄酮类化合物处理后，通过MTT试验检测两种人MM细胞（ARP1、H929）的增殖情况。此外，通过染色和流式细胞术分析细胞周期。通过蛋白质印迹法、转录组RNA测序以及RT-qPCR检测某些蛋白质的差异表达。
结果显示，HKC治疗后MM易感动物似乎得到了保护，生存率延长证明了这一点。此外，9种黄酮类化合物中的4种[金丝桃苷/金丝桃素，HK-2；大麻苷，HK-3；3 - 山柰酚 - 3 - 乙酰基 - 6 - （对香豆酰基） - β - D - 吡喃葡萄糖苷，HK-11；8 - （2'' - 吡咯烷酮 - 5'' - 基） - 槲皮素，HK-B10]诱导了小鼠前成骨细胞MC3T3-E1细胞的分化。另外，两种化合物[异杨梅素，HK-8；槲皮素 - 8 - （2'' - 吡咯烷酮 - 5'' - 基） - 3' - β - D - 吡喃葡萄糖苷，HK-E3]抑制了小鼠Raw264.7细胞中的破骨细胞生成。HK-11直接抑制MM细胞（ARP1和H929）的增殖并诱导G0/G1期细胞周期停滞，这可能涉及抑制β - 连环蛋白，增加IL-6和TNF-α的表达，以及激活成熟的TGF-β1和其他一些代谢途径。
本研究的这些结果表明，HKC的生物活性成分通过促进成骨细胞生成和抑制破骨细胞生成，对MM小鼠的生存发挥保护作用，从而改善骨髓微环境以抑制MM细胞增殖。
