The preparation of chembhu chenduram is a herbal mineral formulation which is a combination of materials like (Copper powder, Nelumbo nucifera(leaf), Sphaeranthus indicus (leaf), Magnolia champaca (flower), Azardirachta indica (flower). This formulation of the herbomineral drug can prevent the cardiovascular disease.
The purification of copper is by the reduction process mediated through green methods. The purified copper mineral is then combined with the medicinal herbals, resulting in the formation of chembhu chenduram Nano herbal Formulation (CC-NhF) Drug.
This formation of chembhu chenduram is then preceded by the toxicological analysis in Zebra fish and animal models (acute and chronic toxicity), in a dose dependant manner at different concentrations. The Minimal inhibition concentration is calculated for analysing the toxicity, then we shall proceed with the characterisation of formulation with different tools like X-ray powder diffraction (XRD) analysis for checking the crystalline nature, size and shape of the particles using Scanning Electron microscopy (SEM) and Transmission electron microscopy (TEM), the quantitative analysis of bioactive compounds (like flavonoids, phenols, terpenoids, tannins, glycosides) can be done using High Performance Thin layer Chromatography (HPTLC) analysis.
The HAECs and U937 cells will be incubated in a medium containing CC-NhF at various concentrations, in a dose-dependent manner. Then to study the dissolution of CC-NhF by monocyte phagocytosis, U937 cells will be incubated with the CC-NhF in different concentrations. Meanwhile, HAECs and U937 cells incubated in the particle-free medium will serve as the controls.
The viability of the HAECs and U937 cells after exposure to CC-NhF will be examined by the Cell counting. Intracellular ROS generation in U937 cells will be determined using the probe 2′,7′-dichlorofluorescein diacetate (DCFH-DA). Intracellular nitric oxide synthesis and secretion in HAECs will be analysed using the probe DAF-FM DA (3-Amino, 4-aminomethyl-2′,7′-difluorescein diacetate).
Inhibitory effect of CC-NhF will be tested on Serum Cardiac biomarkers against doxorubicin-induced cardiac necrosis in rats. We will also check the effect on Isoenzyme pattern of LDH CK, serum biomarkers and Cardiac Biomarkers. Mitochondrial evaluation for its protective effect for CC-NhF against doxorubicin-induced cardiac necrosis in Wistar rats will be analysed. We will check its effect on Serum Metabolites, Mitochondrial Hydrogen Peroxide production, TCA Cycle marker enzymes, Mitochondrial enzymes, Lysosomal marker, DNA, Proteolytic activity on Collagen and Calpain expression on Casein Zymography.