小柴胡汤通过miR-212调控放射性肝损伤的机制

Our previous clinical trials have suggested that xiaochaihu tang could prevent radiation-induced liver disease(RILD). However, little is known about how xiaochaihu tang regulates RILD. It is known that TGF-β1 plays an important role in the pathogenesis of RILD. Moreover, our previous study found that xiaochaihu tang can inhibit the expression of TGF-β1. However, the mechanisms of xiaochaihu tang for the prevention and treatment of RILD remains to be further explored. Therefore, we hypothesized that xiaochaihu tang could contribute to the inhibition of TGF-β1 via the miR-212 pathway, and consequently prevent RILD. To determine whether this hypothesis is valid, we intend to use real time PCR, Western blot, lentiviral vector transfection and RNA interference technology to knockout, overexpress and repress the expression of related genes both in vivo and in vitro. This study may reveal a new mechanism of Traditional Chinese Medicine through which the miR-212/TGF-β1 pathway regulates RILD to provide new insights about the prevention and treatment of RILD.

我们在临床中发现，小柴胡汤能有效防治放射性肝损伤(RILD)的发生，但是其具体机理仍不清楚。已报道TGF-β1在RILD的发生、发展演变中发挥重要作用。我们的前期研究表明，小柴胡汤能明显抑制肝细胞TGF-β1的表达，但其作用机制尚不明了。而软件预测分析显示TGF-β1 是miR-212调控的下游靶基因。为此，我们提出假说：小柴胡汤可能通过miR-212途径下调TGFβ1表达，从而抑制RILD的发生。为了验证这一假说，我们拟采用定量PCR、蛋白印迹、慢病毒载体转染、RNA干扰等技术，通过基因敲除、基因过表达等多种手段上调或下调关键基因的表达，从分子水平、细胞水平、整体水平三个方面，①明确miR-212途径在RILD的发生、发展中的作用；②进一步观察小柴胡汤对该信号途径的影响，③以及对RILD的防治作用。本研究将从miR-212这个新视点丰富中药治疗的疗效机理，为RILD的防治提供新的思路。

放疗引起的放射性肝损伤（RILD）在临床中时有发生，之前研究中，我们发现小柴胡汤能有效防治RILD发生，但具体机制仍未清楚。本研究从细胞水平证实肝细胞癌miRNA-212通过靶向调控TGF-β1基因影响RILD，从动物水平证实肝细胞癌中miRNA-212通过靶向调控TGF-β1基因影响RILD。并研究证实小柴胡汤通过miR-212/ TGF-β1信号通路影响RILD。同时，分析发现hPVT1和uc002mbe.2可联合AFP提高肝细胞癌诊断率。另外，利用类似研究方法发现血清中LOC285194, RP11-462C24.1和 Nbla12061三者结合作为结直肠癌诊断新途径。对RILD的发生发展机制以及临床治疗有重要意义，同时发现的lncRNA标志物有望应用于临床诊断。