Development and Pharmacological Evaluation of a Hepato-Protective Polyherbal Formulation

Abstract

The project was undertaken to develop a hepatoprotective polyherbal formulation comprising methanolic extract of ariel root of Tinospora cardifolia (TC), dried fruits of Carica papaya (CP) and leaf of Justicia gendarussa (JG). The extracts were prepared by Soxhlet’s method and evaluated for their preliminary phytochemicals studies, in-vitro antioxidant and free radical scavenging potentials (DPPH scavenging, ferric ion reduction, hydrogen peroxide scavenging, lipid peroxidation, total phenolic and flavonoid content). The extract of TC has shown excellent antioxidant and free radical scavenging activity, where as JG was least active. All extracts were evaluated for their hepatoprotective activity in-vitro on isolated rat hepatocytes. The trypan blue exclusion assay, determination of GOT, GPT and total protein levels in the cell suspension culture were considered as hepatoprotective index. Due to presence of high amount of total phenolic and flavonoid content, TC has exhibited excellent hepatoprotective activity in-vitro, followed by Carica papaya and Justicia gendarussa. All the extracts were evaluated for their hepatoprotective activity on CCl4 induced hepatotoxicity in rats. The evaluation was done by determining the serum enzymes like, GOT, GPT, ALP and total protein & bilirubin. The wet liver weights as well as histopathological studies were also done as the index of hepatoprotection. The TC has significant hepatoprotective activity in reversing the changes caused by acute dose of CCl4, where as JG and CP possess significant but protection offered by these two were found to be less than the TC. Once the extract were found to be hepatoprotective and possesses antioxidant and free radical scavenging activity, all extracts were mixed in different proportions and six Poly Herbal Preparations (PHP I to VI) were prepared. All six polyherbal preparations were subjected for the in-vitro hepatoprotective activity on isolated rat hepatocytes and evaluated as described above. The best three formulation (PHP II, III and VI), which have shown excellent hepatoprotective activity were selected for further studies. The three polyherbal preparations were evaluated for their hepatoprotective activity on CCl4 induced hepatotoxicity in rats. The activity was assessed by taking serum and non serum parameters as explained above. Out of the three preparations subjected for their hepatoprotective potential, one PHP (PHP III) has produced excellent hepatoprotective activity. The activity was found to more than the individual ingredients and very much similar to the standard hepatoprotective drug Silymarin. The polyherbal preparation, which shown excellent activity was further assessed in drug induced hepatotoxicity. The formulation has reversed the changes caused by the Paracetamol (acute), Anti tubercular (chronic) drugs as well as CCl4 (chronic) induced hepatotoxicity in rats. The evaluation was done by the serum parameters as shown above as well as liver homogenate studies like estimation of SOD, Catalase, Glutathione and lipid peroxidation. The formulation has excellent activity in all the parameters and very much similar with the standard hepatoprotective formulation Liv52. The histopathological studies also revealed the dose dependent hepatoprotective activity of the developed polyherbal formulation. HPTLC profile of the formulation was developed as standardization part of the formulation.