Objective To investigate the effect of ursolic acid(UA) on NOX2/ROS/NLRP3 inflammasome activation in carbon tetrachloride(CCl4)-induced liver fibrosis SD rat,and to observe the improvement of collagen deposition in liver tissues. Methods All rats were randomly divided into 3 group:control group,CCl4model group,UA treatment group. Liver fibrosis model SD rats was established by the CCl4-induced method and half of them was used as UA treatment group. Serum ALT was detected by ALT detection kit.The liver pathology and collagen deposition were ob-served by HE and Sirius-red staining. The mRNA expression of Nox2,Nlrp3,Caspase1,IL-1β in liver tissues was detected by RT-qPCR. The protein expression of NOX2,NLRP3,caspase-1 and IL-1β in liver tissues was detected by Western blot and immunohistochemistry and the ROS generation in liver tissues was detected by DCFH-DA fluores-cence probe. Results Compared with control group,in the CCl4model group,the serum ALT was much higher (P<0.05);the Ishak's fibrosis score and collagen deposition was significantly increased(P<0.05) and mRNA of Nox2, Nlrp3,Caspase1,IL-1β was increased.In addition,both the NOX2,NLRP3,caspase-1 p10 and IL-1β protein expres-sion and ROS level (P<0.05) of CCl4model group were significant increased.Compared with CCl4model group,in the UA treatment group Ishak's fibrosis score,collagen deposition and ALT decreased.Both mRNA expression of the Nox2, Nlrp3,Caspase1,IL-1β and protein expression of NOX2,NLRP3,caspase-1 p10 and IL-1β as well as ROS were signif-icant decreased,but the caspase-1 p45 protein level has no difference among all these groups (P>0.05). Conclusions Ursolic acid attenuates the liver injury and reduces the collagen deposition,which may relate to its inhibitory effects on NOX2/ROS/NLRP3 inflammasome activation to reduce IL-1β releasing.

BACKGROUNDThe domestic prevalence of chronic hepatitis B (CHB) in China is 7.18% in 2006, imposing great societal healthcare burdens. Nucleot(s)ide analogues (NUCs) anti-hepatitis B virus (HBV) therapies are widely applied despite the relatively low rate of seroconversion and high risk of drug-resistant mutation. More effective treatments for CHB deserve further explorations. Combined therapy of NUCs plus Chinese herbal medicine (CHM) is widely accepted in China, which is recognized as a prospective alternative approach. The study was primarily designed to confirm the hypothesis that Tiaogan-Yipi Granule (, TGYP) or Tiaogan-Jianpi-Jiedu Granule (, TGJPJD) plus entecavir tablet (ETV) was superior over ETV monotherapy in enhancing HBeAg loss rate.

METHODSThe study was a nationwide, large-scale, multi-center, double-blind, randomized, placebo-controlled trial with a designed duration of 108 weeks. A total of 16 hospitals and 596 eligible Chinese HBeAg positive CHB patients were enrolled from November 2012 to September 2013 and randomly allocated into 2 groups in 1:1 ratio via central randomization system: experimental group (EG) and control group (CG). Subjects in EG received CM formulae (TGYP or TGJPJD, 50 g per dose, twice daily) plus ETV tablet (or ETV placebo) 0.5 mg per day in the first 24 weeks (stage 1), and CHM granule plus ETV tablet (0.5 mg per day) from week 25 to 108 (stage 2). Subjects in CG received CHM Granule placebo plus ETV tablet (0.5 mg per day) for 108 weeks throughout the trial. The assessments of primary outcomes (HBV serum markers and HBV-DNA) were conducted by a third-party College of American Pathologists (CAP) qualified laboratory. Adverse effects were observed in the hospitals of recruitment.

DISCUSSIONThe study was designed to compare the curative effect of CM plus ETV and ETV monotherapy in respect of HBeAg loss, which is recognized by the European Association for the Study of the Liver as "a valuable endpoint". We believe this trial could provide a reliable status for patients' "journey" towards durable responses after treatment discontinuation. The trial was registered before recruitment on Chinese Clinical trial registry (No. ChiCTR-TRC-12002784, Version 1.0, 2015/12/23).