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小柴胡汤含药血清通过 JAK2/STAT3 信号通路对 HepG2.2.15 细胞的影响。

Effect of modified Xiaochaihu decoction‑containing serum on HepG2.2.15 cells via the JAK2/STAT3 signaling pathway.

机构信息

College of Chinese Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, P.R. China.

出版信息

Mol Med Rep. 2017 Nov;16(5):7416-7422. doi: 10.3892/mmr.2017.7561. Epub 2017 Sep 20.

Abstract

The present study aimed to investigate the possible mechanisms underlying the effect of modified Xiaochaihu decoction (mXCHD) in the treatment of chronic hepatitis B (CHB). Patients with CHB, in addition to liver stagnation and spleen deficiency syndrome were randomly assigned to receive either Chinese (mXCHD) or western (entecavir) treatment, with 30 cases in each group. Serum was collected following treatment with mXCHD or entecavir for 7 days. A healthy group of 30 individuals was also included. HepG2.2.15 cells were cultured in vitro and randomly divided into four groups: Healthy; entecavir‑treated; 10% mXCHD‑treated; and 20% mXCHD‑treated. The HepG2.2.15 cells in the four groups were treated with either serum from the healthy volunteers, entecavir‑containing serum, or mXCHD‑containing serum at different concentrations (10 or 20%, respectively). Following treatment with the corresponding serum, cell proliferation was examined using an MTT assay, and the expression of hepatitis B surface antigen (HBsAg) in the cell supernatant was detected using an enzyme‑linked immunosorbent assay. The mRNA and protein expression levels of Janus kinase (JAK)2 and signal transducer and activator of transcription (STAT)3 were measured using reverse transcription‑quantitative polymerase chain reaction and western blot analyses, respectively. The results indicated that the most effective treatment for the promotion of HepG2.2.15 cell proliferation was a 20% concentration of mXCHD serum. The expression of HBsAg was significantly decreased in the groups treated with 10 and 20% mXCHD 48 h following intervention (P<0.01). The mRNA and protein expression levels of STAT3 in the 20% mXCHD serum group were significantly increased, compared with those in the healthy group (P<0.01 and P<0.05, respectively), whereas no significant difference was observed in the expression of JAK2 among the four groups. These results indicated that mXCHD suppressed the hepatitis B virus, and treatment of the cells with mXCHD‑containing serum promoted HepG2.2.15 cell proliferation via modulating the expression of STAT3, which may contribute to the clinical efficacy of mXCHD against CHB.

摘要

本研究旨在探讨加味小柴胡汤(mXCHD)治疗慢性乙型肝炎(CHB)的作用机制。将 CHB 患者(兼有肝郁脾虚证)随机分为中药组(mXCHD)和西药组(恩替卡韦),每组 30 例。治疗 7 天后收集 mXCHD 或恩替卡韦治疗后的血清。同时纳入 30 例健康人群作为健康组。体外培养 HepG2.2.15 细胞,随机分为四组:健康组、恩替卡韦处理组、10% mXCHD 处理组和 20% mXCHD 处理组。将四组 HepG2.2.15 细胞分别用健康志愿者血清、含恩替卡韦的血清和不同浓度(分别为 10%和 20%)的 mXCHD 血清处理。用相应的血清处理后,通过 MTT 法检测细胞增殖情况,用酶联免疫吸附试验检测细胞上清液中乙型肝炎表面抗原(HBsAg)的表达。用逆转录定量聚合酶链反应和蛋白质印迹分析分别检测 Janus 激酶(JAK)2 和信号转导和转录激活因子(STAT)3 的 mRNA 和蛋白表达水平。结果表明,促进 HepG2.2.15 细胞增殖最有效的治疗方法是 20%浓度的 mXCHD 血清。干预后 48 小时,10%和 20% mXCHD 处理组 HBsAg 的表达明显降低(P<0.01)。与健康组相比,20% mXCHD 血清组 STAT3 的 mRNA 和蛋白表达水平显著增加(P<0.01 和 P<0.05),而四组 JAK2 的表达无明显差异。这些结果表明,mXCHD 抑制乙型肝炎病毒,用含 mXCHD 的血清处理细胞可通过调节 STAT3 的表达促进 HepG2.2.15 细胞增殖,这可能有助于 mXCHD 治疗 CHB 的临床疗效。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc57/5865873/119bbb133d45/mmr-16-05-7416-g00.jpg

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