Anti-hepatic Fibrosis Role Of Recombinant Human Kallistatin Protein

Hepatic fibrosis refers to a pathological process in which the synthesis ofextracellular matrix is increased and degradation decreased, leading to liverconnective tissue abnormal proliferation. Anti-hepatic fibrosis drugs at this stage areuniversally of a single target with a poor specificity. The effect of a single agent fortreatment of hepatic fibrosis is not optimistic. Thus, the research and development foranti-hepatic fibrosis drugs with multiple targets, the sought for combinations of drugswith synergistic anti-hepatic fibrosis functions, become new trends of anti-hepaticfibrosis study.This paper chose Kallistatin, a multifunctional molecule with antioxidant,anti-angiogenesis, vasodilator, anti-inflammatory activities, to achieve themulti-target regulation of hepatic fibrosis and verified the anti-hepatic fibrosis role ofKallistatin through CCl4caused mice hepatic fibrosis model.Firstly, by the preparation and purification of recombinant human Kallistatinprotein through Pichia pastoris shake flask fermentation system, we got the highestyield of14mg/L in shake flask culture. The purified recombinant human Kallistatinprotein had a molecular weight of58kD with98%purity，which provided thematerial for the following animal studies.Secondly, the animal study found a significant therapeutic effect of recombinanthuman Kallistatin protein on CCl4caused hepatic fibrosis mice. When compared withmodel group the results showed that:①Treatment group mice had a higher survivalrate and a relatively normal liver lobule structure with little hepatic cells necrosis andinflammatory cells infiltration.②The expressions of α-SMA and TGF-β₁intreatment group were significantly reduced both in expression range and level.③Asliver function indicators, ALT, AST serum contents of treatment group wereconspicuously reduced and ALB dramatically boosted.④As fibrosis markers, HA,C-III, LN serum contents were significantly reduced.⑤As liver tissue oxidationindex, MDA, Hyp serum contents of treatment group were significantly reduced andSOD significantly elevated.This paper is the first demonstration of the remarkable inhibitory role of the recombinant human Kallistatin protein on inflammation, oxidative stress and collagensynthesis in CCl4caused hepatic fibrosis mice livers. This study provides a theoreticalbasis for the exploration of Kallistatin anti-fibrosis activity through multi-channels.