Phase Preparation of Xiao-chai-hu Decoction and its Pharmacodynamics of Acute Liver Injury
Authors: Su Hongbing , Lei Hong , Cao Ling , Zhou Xiaoying , Liu Yumeng , LI Ying , Yan Lili , Lv Shaowa , Wang Yanhong , Guan QingXia 
Published: August 2024

Background:
Self-emulsifying nano-phase of traditional Chinese medicine are a research
hotspot. Xiao-Chai-Hu decoction is a commonly used compound decoction in clinical practice,
which is of great research significance. The aim of this study was to isolate and characterize
the self-emulsifying nano-phase and other phases of Xiao-Chai-Hu decoction, and to study the effects
of each phase on acute liver injury.

Methods:
The liquid medicine was prepared employing centrifugation followed by dialysis. Single-
factor investigation methodology was utilized to optimize the preparation parameters for both
phases. Characterization of the formulated phase involved analyses such as surface morphology assessment,
measurement of nanoparticle size and Zeta potential using an analyzer, observation of
the Tyndall effect, conducting diffusion and dilution tests, examination under a microscope, and
structural visualization via transmission electron microscopy (TEM). Furthermore, an acute liver
injury model was established in rats through intraperitoneal injection of D-Galactosamine (D-Gal-
N). To assess hepatic function and oxidative stress status, serum levels of alanine aminotransferase
(ALT) and aspartate aminotransferase (AST), superoxide dismutase (SOD) activity, and
malondialdehyde (MDA) content in liver tissue were quantified. The liver coefficients for each
group were calculated as an additional parameter. For histopathological evaluation, liver tissue sections
from the experimental group were stained with Hematoxylin and Eosin (H&E) and examined
microscopically under light conditions. These revisions aim to enhance clarity, correct minor
grammatical errors (such as capitalization of “HE” to “H&E”), and ensure a smoother flow of
information without altering the scientific content of your original text.

Results:
Successful establishment and separation of four distinct phases were achieved, including
the self-emulsifying nano-phase, precipitation phase, suspension phase, and true solution phase.
The self-emulsifying nano-phase was characterized as spherical particles with an average diameter
of approximately 100 nm. Pharmacodynamic assessments revealed that both Xiao-Chai-Hu decoction
and its self-emulsifying nano-phase significantly reduced liver coefficients and alanine aminotransferase
(ALT) levels compared to controls (P<0.05). However, no statistically significant differences
were observed in regards to aspartate aminotransferase (AST) concentrations, malondialdehyde
(MDA) content, or superoxide dismutase (SOD) activity between the treatment groups
and control (P>0.05). These findings indicate that both Xiao-Chai-Hu decoction and its self-emulsifying
nano-formulation ameliorated D-GalN-induced acute liver injury, albeit without statistically
distinguishable efficacy between them (P>0.05).

Conclusion:
The presence of a self-emulsifying nano-phase within Xiao-Chai-Hu decoction is
confirmed, and this nano-phase emerges as a therapeutically efficacious component in mitigating
acute liver injury.