Abstract:
Various studies show that on average, Hepatitis C virus (HCV) prevalence in Pakistan by
percent in adult population is 11.55%. Most of the people diagnosed with HCV infection
are aged around 25-35 with ever increasing incidence, around the globe. Our study
highlights critical aspects of chronic HCV pathology and its effect on the Immune system.
Peripheral Blood Mononuclear cells, specifically CD8+ T cytotoxic lymphocytes and
CD4+ T helper cells endorse a hyper-responsive state owing to the levels of viremia and
antigenemia caused by chronic Hepatitis C virus. Consequently, these cells become highly
dysfunctional. It is well documented that HCV leads to oxidative stress. It also
dysregulates antioxidant defense systems. High levels of reactive oxygen species (ROS)
lead to oxidation and damage of cellular proteins and organelles, as a result Peripheral
Blood Mononuclear cells (PBMCs) become dysfunctional. HCV specific lymphocytes
become exhausted due to constant epitope signaling caused due to HCV specific antigens
as well as DNA damage caused by high levels of oxidative stress caused in the chronic
HCV infection. This leads to the apoptosis of PBMCs via FAS/FAS-L mediated (extrinsic)
and intrinsic pathways respectively. Sulforaphane, which is a naturally derived anti inflammatory and cytoprotective phytochemical can reverse the oxidative stress by
activating the NRF2 signaling pathway that can reverse the apoptosis of exhausted virus
specific PBMCs. In our study PBMCs were isolated, cultured and treated by optimal doses
of Sulforaphane. Cell viability assays were performed to know whether Sulforaphane
(SFN) plays a pivotal part in reversing the anti-oxidant stress, cellular exhaustion and
apoptosis. The changes at the genetic level are marked by performing RT-PCR, to detect
the increased expression of genes involved in anti-oxidant defense systems. The reversal of
these stressors leads to enhanced cell survival, reversal of apoptosis, proliferation and
improved functionality required to ward off viral load. Cross validating assays were
performed using computational biology tools, including molecular docking and GRIND
(advanced alignment independent 3D QSAR) to further validate our findings.
