Objective <p>This study aimed to investigate the effects of recombinant plasmid heat shock protein 47 small interfering ribonucleic acid (HSP47-siRNA) on fibroblast proliferation and collagenous fibre synthesis in the filtering channel after trabeculectomy in rabbit.</p> Methods <p>The recombinant plasmid HSP47-siRNA was constructed successfully to extract sufficient recombinant plasmids. Furthermore, an animal model of anti-glaucoma surgery in rabbit eyes was created to establish the HSP47-siRNA group, empty vector group, mitomycin C (MMC) group and normal saline group. This study further observed the postoperative eye condition, measured the intraocular pressure and carried out histological and immunohistochemical staining analysis.</p> Results <p>At different time points after surgery, there were no significant differences in the comparison of filtering bleb morphology, conjunctival congestion, corneal oedema, inflammation and depth of anterior chamber or lens opacity among the HSP47-siRNA, empty vector, MMC and normal saline groups. Within 7&#xa0;days after surgery, filtering blebs in all groups were bulged and diffused, with no statistically significant difference (<i>p</i> &gt; 0.05). On day 15 and day 30 after surgery, the upper filtering bleb gradually decreased in the empty vector group and the normal saline group compared with the HSP47-siRNA group (<i>p</i> &lt; 0.01). On day 30 after surgery, flat and pale scar tissues were observed in the surgical area of the empty vector and normal saline groups, whereas functional blebs were still observed in the HSP47-siRNA and MMC groups. Within 7&#xa0;days after surgery, the intraocular pressure significantly decreased in each group, with a statistically significant difference compared with that before surgery (<i>p</i> &lt; 0.01).</p> Conclusion <p>HSP47-siRNA exhibits application potential concerning its effects on anti-scarring after glaucoma filtration surgery in rabbit.</p>