# Hepatoprotective Effects and Antioxidant Properties of a Herbal Detoxifying Formula Against Chlorpyrifos-Induced Toxicity in Sprague-Dawley Rats

## TL;DR

A herbal formula from Thailand protects rat livers from pesticide damage by reducing oxidative stress and improving liver function.

## Contribution

The study scientifically validates a traditional Thai herbal formula's hepatoprotective and antioxidant effects against chlorpyrifos toxicity.

## Key findings

- Formula 04 reduced oxidative stress markers and preserved liver tissue in rats exposed to chlorpyrifos.
- The formula restored acetylcholinesterase activity and increased antioxidant enzyme levels in treated rats.
- Formula 04 showed the highest antioxidant and pro-apoptotic activity among seven tested formulations.

## Abstract

Exposure to organophosphate pesticides, such as chlorpyrifos, poses a significant health risk in agricultural communities due to its harmful effects on the liver and nervous system. In northern Thailand, local populations traditionally use herbal remedies for detoxification, but scientific evidence supporting their efficacy is limited. This study evaluated a herbal detoxifying formula (Formula 04) containing Thunbergia laurifolia and Embelia sessiliflora, two medicinal plants traditionally used to alleviate pesticide-related symptoms. In vitro, the formula exhibited strong antioxidant activity and induced apoptosis in hepatic stellate cells, which are associated with liver fibrosis. In vivo, rats exposed to chlorpyrifos were treated with the formula using a traditional alternating-dose regimen over 18 days. Treated animals showed improved liver function, restoration of endogenous antioxidant defenses, reduced lipid peroxidation, and preservation of liver tissue architecture. No adverse effects or toxicity were observed. These results support the finding that this herbal formula is safe and effective for pesticide detoxification and provide scientific validation for its use in traditional Thai folk medicine.

Chlorpyrifos (CPF), a widely used organophosphate pesticide, induces hepatotoxicity primarily through oxidative stress, acetylcholinesterase (AChE) inhibition, and inflammatory responses. This study evaluated the hepatoprotective potential of detoxifying formulations prepared from four medicinal plants, Thunbergia laurifolia, Embelia sessiliflora, Morinda angustifolia, and Thunbergia coccinea, in various ratios. Among these, a formulation composed of T. laurifolia and E. sessiliflora (1:1; Formula 04) showed the highest activity, with rosmarinic acid identified by compact mass spectrometry (CMS). Formula 04 demonstrated the greatest antioxidant and pro-apoptotic potential among the seven tested formulations, as confirmed by in vitro DPPH and superoxide radical assays and apoptosis assays in hepatic stellate LX-2 cells. In vivo, Sprague–Dawley rats exposed to CPF (16 mg/kg/day, oral gavage) received Formula 04 orally 30 min prior to CPF exposure in 6 cyclic dosing regimen for 18 days. The treatment restored AChE activity, increased superoxide dismutase levels, reduced glutathione levels, and decreased malondialdehyde, indicating attenuation of oxidative stress. Serum AST levels were significantly reduced compared with the CPF group, and histopathology revealed that the hepatic architecture was preserved. These findings demonstrate that Formula 04 exerts hepatoprotective effects against CPF-induced toxicity via redox regulation and cellular protection mechanisms, demonstrating its potential as a natural detoxifying agent.