扶正化瘀片调节肝纤维化miR-122基因相关因子表达研究

Expression of miR-122-related factors of liver fibrosis regulated by Fuzheng Huayu Tablet

目的探讨扶正化瘀片联合抗病毒治疗肝纤维化患者的软坚散结作用,以及是否与微小基因-122/白细胞介素-10/活性氧(miR-122/IL-10/ROS)通路调节作用有关。方法选取2015年8月至2016年8月慢性肝炎肝纤维化患者85例,随机分为试验组(n=45)和对照组(n=40)。对照组给予恩替卡韦(0.5 mg/d)以及甘草酸单铵(0.1 g/d),试验组在此基础上同时给予扶正化瘀片(4.5 g/d),疗程为48周。治疗结束后,常规病理评价ISHAK纤维化评分;采用ELISA法测定血清超氧化物歧化酶(SOD)、活性氧(ROS)活性;RT-PCR测定肝组织治疗前后各组miR-122和IL-10mRNA的表达;免疫比浊法测定血清肝纤维化炎症因子四项,即III型前胶原(PCIIINP)、IV型胶原(IV-C)、透明质酸酶(HA)、层粘连蛋白(LN)的含量。结果治疗前对照组、试验组Ishak纤维化评分无显著差异,治疗后对照组、试验组ISHAK纤维化评分均较治疗前下降,试验组治疗改善纤维化ISHAK评分优于对照组(P<0.05)。对照组、试验组治疗后血清肝纤维化四项指标较治疗前均下降,其中试验组PCIIINP、HA下降更为明显,与对照组比较有统计学差异(P<0.05)。对照组、试验组治疗前ROS、SOD无差异,治疗后2组ROS含量较治疗前均降低,治疗后试验组抑制ROS效果优于对照组(P<0.05),试验组治疗后SOD含量增加(P<0.05)。对照组、试验组治疗后IL-10和miR-122 mRNA表达较治疗前均有降低,试验组较对照组更明显(P<0.05)。结论扶正化瘀片直接降低胶原蛋白和HA水平,降解细胞外基质异常沉积,调节胶原蛋白动态平衡。扶正化瘀片有效地拮抗IL-10、-122基因表达,减轻细胞间质炎症反应,控制肝纤维化进展。

Objective To investigate softening-hardness dissipating-binds effects of Fuzheng Huayu （reinforcing-healthy qi resolving-stasis ） Tablet combined with anti-virus therapy on liver fibrosis, and relationship between these effects and regulation of pathway of miR-122-interleukin-10-reactive oxygen species （miR-122/IL-10/ROS）. Methods The patients with chronic hepatitis and liver fibrosis （n = 85） were chosen from Aug. 2015 to Aug. 2016, and then divided randomly into test group （n =45） and control group （n = 40）. The control group was treated with entecavir （0.5mg/d） and monoammonium glycyrrhizinate （0.1 g/d）, and test group, additionally with Fuzheng Huayu Tablet （4.5 g/d） for 48 weeks. After treatment, ISHAK fibrosis score was reviewed through routine pathology, and activities of serum superoxide dismutase （SOD） and ROS were detected by using ELISA. The expressions of miR-122 mRNA and IL-10 mRNA were detected by using RT-PCR, and content of serum inflammatory factors of liver fibrosis, including proeollagen III N-terminal peptide （PCIIINP）, type IV collagen （IV-C）, hyaluronidase （HA） and laminin （LN）, were detected by using immunoturbidimetry. Results ISHAK fibrosis score had no significant difference between 2 groups before treatment, and decreased in 2 groups after treatment, which was superior in test group to that in control group （P 〈 0.05）. The content of serum inflammatory factors of liver fibrosis all decreased in 2 groups after treatment, and the decreases of PCIIINP and HA were more significant in test group compared with control group （P 〈 0.05 ）. The activities of [lOS and SOD had no difference in 2 groups before treatment, and ROS decreased in 2 groups after treatment. The inhibitory effect on [lOS was better in test group than that in control group after treatment （P 〈 0.05 ） , and SOD increase was effectively relieved in test group after treatment （ P 〈 0.05）. The expressions of miR-122 mRNA and IL-10 mRNA all decreased in 2 groups after treatment, which was more significant in test group compared with control group （P 〈 0.05 ）. Conclusion Fuzheng Huayu Tablet reduces directly the levels of collagen and HA, degrades abnormal deposition of extracellular matrix, and regulates dynamic balance of collagen, and it antagonizes effectively the genetic expressions of IL-10 and miR-122, relieves inflammatory disorders and controls progress of histology.