IN VITRO STUDY OF INHIBITION OF PMCA4 GENE EXPRESSION IN SPERMATOGENESIS WITH CHITOSAN NANOPARTICLES CONTAINING PHYLLANTHUS NIRURI USING QPCR ASSAY

• Main Author: Arita, Ditri
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/78404
• Institution: Institut Teknologi Bandung

Phyllanthus niruri (P. niruri) is commonly used in herbal medicine because it has many pharmacological activities including immunomodulatory, antidiabetic, and antihypertensive. In terms of safety, this plant has unwanted effects, such as hypoglycemia and hypotension. In male rats, the administration of this extract showed an effect on spermatogenesis, such as accumulation of fluid in the testes, abnormalities in sperm, shrinkage and reduction of tubular diameter, and increase in organ weight which has the potential to cause infertility in males. This study aims to determine the effect of P. niruri extract, empty chitosan nanoparticles, and P. niruri extract encapsulated in chitosan nanoparticles on spermatogenesis through the Pmca4 gene using the qPCR method in vitro. Chitosan nanoparticles were prepared using the ionic gelation method with the addition of STPP as a polyanion. The particle size obtained for empty chitosan nanoparticles was 154,30 with a polydispersity index of 0,30. Meanwhile, the particle size for chitosan nanoparticles containing P. niruri extract was 167,56 nm with a polydispersity index of 0,278. The zeta potential of empty chitosan nanoparticles and chitosan nanoparticles containing P. niruri were +34,01 mV and +37,19 mV, respectively. Formulation of chitosan nanoparticles containing P. niruri extract was prepared in nine concentration series and then transfected into TM4 cells, RNA cells were isolated and reverse transcription was performed to obtain cDNA. Based on the tests, it was found that chitosan nanoparticles containing P. niruri extract at concentrations of 250, 500, 1000, and 2000 ppm significantly inhibited Pmca4 gene expression, but no significant inhibition occurred after administration of empty chitosan nanoparticles or P. niruri extract. The presence of Pmca4 gene inhibition causes decreased sperm motility and infertility in men.