Mechanistic exploration of the activities of poly(lactic-co-glycolic acid)-loaded nanoparticles of betulinic acid against hepatocellular carcinoma at cellular and molecular levels

Abstract

The effectiveness of betulinic acid (B) and PLGA loaded nanoparticles of B (Bnp) against hepatocellular carcinoma (HCC) was established and reported earlier. In continuation of our previous report, the present study described the molecular mechanisms of their antineoplastic responses. In this context, the antineoplastic properties of both B and Bnp were evaluated on DEN-induced HCC rat model. The quantitative real-time polymerase chain reaction and western blot analyses revealed that HCC was developed through lower expressions of e-NOS, BAX, BAD, Cyt C and higher expressions of i-NOS, Bcl-xl, Bcl-2. B and Bnp normalised the expressions of these apoptogenic markers. Particularly, both activated i-NOS and e-NOS mediated Bcl-2 family proteins→CytC→Caspase 3 and 9 signalling cascades. The ¹H-NMR-based metabolomics study also demonstrated that the perturbed metabolites in DEN-induced rat serum restored to the normal level following both treatments. Moreover, the antineoplastic potential of Bnp was found to be comparable with the marketed product, 5-flurouracil.

Keywords:

• PLGA nanoparticles
• betulinic acid
• hepatocellular carcinoma
• Bcl-2
• Bcl-xl and BAX proteins
• ¹H-NMR-based metabolomics

Disclosure statement

The authors declare that they do not have any conflict of interests.

Additional information

Funding

Dr. Sudipta Saha thanks UGC, India, for UGC-MRP grant [Project no. 42–680/2013(SR)] and DST, India for DST-SERB project [Ref. No. DST/SB/EMEQ-320/2014]. The authors also express their sincere gratitude to CBMR, Lucknow for providing the NMR facilities. Mr. Pranesh Kumar acknowledged DST India for the INSPIRE Fellowship [DST/INSPIRE Fellowship/2016/IF160364].