Review on the Toxic Effects of Radix Bupleuri

REVIEW
Research paper
Curr Opin Complement Alternat Med 1:1, 3-7; January/February 2014; © 2014 STM Publishing
Review on the Toxic Effects of Radix Bupleuri
Yamin LIU, Zongyang LI, Xinmin LIU, Ruile PAN
Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education, Institute of Medicinal Plant
Development, Peking Union Medical College, Beijing 100193, China
Summary
The present paper reviewed the physical basis, influencing factors, and mechanism of the toxic effects of Radix Bupleuri to provide
reference for clinical applications. Related studies regarding the toxicity of Radix Bupleuri published locally and internationally
in the past 10 years were collected and summarized. We found that Bupleurum saikosaponins and essential oil are the main toxic
components. The main organ prone to toxicity is the liver, and hepatic damage is mainly related to multichannel oxidative injury.
Different kinds, processing methods, and extraction methods elicit various effects on Radix Bupleuri toxicity. As such, Radix Bupleuri
toxicity has been gradually recognized in clinical applications. The extent of toxicity is related to Bupleurum saikosaponin and volatile
oil contents.
Curr Opin Complement Alternat Med 2014; 1:3-7
Key words
saikosaponin; Radix Bupleuri; toxicology research; mechanism of hepatic lesion; hepatotoxicity; summary
Radix Bupleuri is one of the commonly used traditional Chinese
medicines in Chinese history. Radix Bupleuri was firstly recorded
in Shen Nong’s Herbal Classic and ranked as a top grand. As
described in the Chinese pharmacopoeia, Radix Bupleuri is derived
from the dried roots of Bupleurum chinense DC or Bupleurum
scorzonerifolium Willd. Radix Bupleuri has been used traditionally
to relieve fever, enhance the texture of the liver, and cure uterine
prolapse, dysmenorrhea, and rectocele.1-10 Radix Bupleuri has
also been used to treat liver diseases, including chronic and viral
hepatitis.11 This plant also exhibits scavenging activity against
reactive oxidative species. Therefore, Bupleuri Radix is frequently
used as a major medicinal herb in traditional Chinese medicine
(TCM).
Thus far, saikosaponins, which are the main components of
Radix Bupleuri, have been used to treat liver fibrosis, tumor,
and inflammation, as well as regulate the immune system; these
components have been the focus of research progress.12 Bupleurum
volatile oil is obtained by steam distillation and mainly used to
treat influenza, cold, tonsillitis, respiratory infections, and other
kinds of fever caused by diseases; which exhibiting an accurate
curative effect.13
Although Radix Bupleuri is not listed as a toxic material in
materia medica during production, this plant produces side effects
in practical applications; Ye Tianshi in Qing dynasty proposed the
“Bupleurum chinense Robbing Hepatic Yin” as the main theory.14,15
Correspondence to: Ruile PAN, Key Laboratory of Bioactive Substances and
Resources Utilization of Chinese Herbal Medicine, Ministry of Education, Institute
of Medicinal Plant Development, Peking Union Medical College, Beijing 100193,
China; Email: [email protected]
Submitted: 09/07/2013; Revised: 13/08/2013; Accepted: 01/09/2013
DOI: 10.7178/cocam.10
www.cocam.org
In recent years, clinical reports on drug-induced hepatic
damage have been presented, particularly the largest scale of
poisoning incidents caused by B. chinense in Japan.16,17 This paper
reviews the component that elicits toxicity and other influencing
factors. The mechanism by which toxic effect is induced is also
described, thereby providing reference for clinical applications.
Different toxic effects of Radix Bupleuri
Low toxicity occurs when this plant is administered at
recommended dosages in pharmacopoeia; however, a long-term
high dosage causes significant toxicity on the kidney, liver, and
blood system.18
Major symptoms include transaminase lifts, hepatitis, and
jaundice when toxicity causes Radix Bupleuri-induced liver
damage; liver biopsy also indicates acute liver damage; however,
withdrawal after a specific period, liver functions can return to
normal levels.19-22
Japanese scholars reported that Xiao Chaihu Tang may
cause interstitial pneumonia and even death when this drug is
administered to patients with long-term oral chronic hepatitis
and liver cirrhosis.16,23 In addition, liver damage caused by this
prescription can be attributed to protoplasmic toxin.24 With
the increasing use of Bupleurum extract in clinical applications,
additional adverse reactions, such as liver damage, allergic reaction,
anaphylactic shock, and kidney failure, have been reported.13,25-28
In previous studies, the essential oil of Radix Bupleuri is
intravenously injected to mice and rats, causing acute toxic damage
with the following symptoms: agitation; asynchronous state;
higher heart rate; fast breathing; and continuous twitching.29,30 At
a high dose, extracted Radix Bupleuri saponins used as a longterm feed of rats can also cause significant liver damage manifested
Curr Opin Complement Alternat Med
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Research paper
as hepatocyte organic lesion and liver function changes, as well as
hepatocyte death, which occurs in a specific manner of quantity,
time, and toxicity.31-35
Chemical constituents of toxins
Phytochemical studies of approximately 50 Bupleurum species have
contributed to the isolation and identification of approximately
250 natural compounds from the major phytochemical classes. In
general, chemical constituents and qualities of biological species
vary; however, the majority of chemical constituents, which are
isolated from different species, belong to lignins, terpenoids,
phenolics,36,37 and polyacetylenes. In addition to the main
compounds, minor components, including phenylpropanoids,
polysaccharides, and a few alkaloids, have also been isolated.38
From this view, the current knowledge on the chemical structures
of compounds was summarized in this paper.
Essential oils are a common characteristic of the Apiaceae
family, and the representatives of 20 Bupleurum species produce
essential oils, such as hexanal, heptanal, (E)-2-nonenal, (E,E)2,4-decadienal, hexanoic acid, heptanoic acid, octanoic acid, and
hexadecanoic acid (Fig. 1).39-44
Studies have shown that saikosaponins and volatile oil are
the main toxic components and active ingredients. The main
saikosaponins are divided into saikosaponin A, saikosaponin C,
saikosaponin D, saikosaponin B1, saikosaponin B2, saikosaponin E,
and saikosaponin F; the main sapogenins are aglycone F, aglycone
E, aglycone G, and alpha pinasterol; in addition, Δ7-stigmasterol,
Δ22-stigmasterol, stigmasterol, adonitol, and angelicin are mainly
found in the volatile oil of Radix Bupleuri (Fig. 2).45-48
However, saikosaponins are also hemolytic; the intensity
of hemolysis can be expressed as follows: saikosaponin D
> saikosaponin A > saikosaponin B1 > saikosaponin B2 >
saikosaponin C.1,29 Different components can cause liver damage
in rats; studies have also suggested that liver damage caused by
ethanol extracts is higher than that caused by water extracts.49,50 In
other studies, purified saikosaponins are used as a long-term feed
of mice and rats; at a high dose, these saikosaponins cause evident
liver lesion.51,52 However, purified saponins eluted using hydrous
alcohol cause long-term liver tissue damage.53 The essential oil
of B. longiradiatum Turcz. causes higher toxicity because of toxic
components, namely, bupleurotoxin and 14-acetoxybupleurotoxin
(Fig. 3).54,55
Factors eliciting toxic effects
Many factors of Radix Bupleuri elicit different toxic effects. These
factors include original plant, processing methods, extracting
methods, and metabolism (Table 1).
Influence of original plant
Original plant varieties are complex, except B. scorzonerifolium
Willd and B. chinense DC in the pharmacopeia provision, as well
as B. smithii var. parvifolium, B. marginatum var. stenophyllum,
B. marginatum Wall. ex DC, B. bicaule Helm in Mem, B. smithii
Wolff, and B. longiradiatum Turcz.56,57 Among these species, B.
longiradiatum Turcz elicits the highest toxicity among Bupleurum
4
plants. In the early 1970s, a severe poisoning caused by a drug
obtained from B. longiradiatum Turcz occurred in MuLan Xian
County, Heilongjiang Province, China.58
Ultraviolet
spectrometry,
high-performance
liquid
chromatography, and classical animal acute toxicity have been
conducted to probe the relationship between total saikosaponins
and acute toxicity. Result shows that B. sinenses DC elicits greater
toxicity than B. scorzonerifolium Willd; in mice fed with total
saikosaponins, the extent of toxicity is correlated with the content
at a certain degree.59
Influence of processing
Sun53,60 compared the SSa content of B. sinenses DC that elicits
acute toxicity after different processing techniques were performed,
and the following results were obtained: raw products > products
fried with vinegar > products fried with wine > products with
honey processing > products fried without additional ingredients;
after the mice were fed with each processed product, the degree
of acute toxicity was observed as follows: raw product > products
with vinegar processing > products with wine processing >
products fried without additional ingredients > products with
honey processing. The SSa content also elicited acute toxicity
after different processing techniques were performed using B.
scorzonerifolium Willd; the result is summarized as follows: raw
products > products with honey processing > products with
vinegar and wine processing > products fried without additional
ingredients; after the mice were fed with each processed product,
the degree of acute toxicity was observed as follows: raw products
> products with vinegar processing > products without additional
ingredients > products with wine processing > products with
honey processing. The changes in SSa content and acute toxicity
among different processing products are the same. The SSa
content in products with processed honey is lower than that
in other processed products; this finding indicates that honey
processing can increase the dosage range at which the drug is safe.
The toxicity of B. longiradiatum can be reduced by drying
under the shade, stoving, stewing, and soaking; toxicity can be
eliminated by drying in hot air. Experiments have also shown that
the toxicity of B. longiradiatum is less than that of the common
medicinal samples; therefore, B. longiradiatum can provide the
basis of the medicinal importance of reusing B. longiradiatum.61
Influence of extracting methods
The enrichment degree is different among different preparation
methods of saikosaponins and mouse acute toxicity. High amounts
of saikosaponins induce greater toxicity than low amounts of
saikosaponins.
Another study has demonstrated that different degrees of acute
toxicity are observed in mice treated with ethanolic elution for 14
days. Such variation is attributed to the concentration of ethanolic
elution containing different SSa contents that can be used as a
toxicity index; the acute toxicity in mice at 70% ethanolic elution
is greater than that at 50%, 30%, 80%, and 95% ethanolic
elution.62
The solvent extracts of B. chinense DC and B. scorzonerifolium
Willd vary at different intensities of acute toxicity. For instance,
toxicity is higher than that of B. scorzonerifolium Willd when mice
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n-hexanal
heptanoic acid
hexanoic acid
bupleurotoxin
octanoic acid
acetyl-bupleurotoxin
hexadecanoic acid
Figure 3 Structures of bupleurotoxin and acetyl-bupleurotoxin
Figure 1. Main structures of Radix Bupleuri essential oils
R1
R2
R3
Saikosaponin B1 β-OH OH β-D-glu-(1→3)-β-D-fuc
Saikosaponin M
H
OH β-D-glu-(1→3)-β-D-fuc
R1
Saikosaponin A β-OH
Saikosaponin D α-OH
Saikosaponin C β-OH
R2
R3
OH β-D-glu-(1→3)-β-D-fuc
OH β-D-fuc
H β-D-glu-(1→6)-[α-L-rha-(1→4)]-β-D-glu
Figure 2. Structures of the main saikosaponins in Radix Bupleuri
Table 1. Factors influencing the toxicity of Radix Bupleuri
Influencing factors
Original plant
Processing methods
Specific performance
Varieties are complex; toxic effects of different species vary from one another.
Methods include wine processing, wine fries, vinegar processing, vinegar steaming, honey processing, and ginger processing.
Honey processing can lower the toxicity and improve the safety of clinical use.
Extracting methods
Metabolites
Different extract solvents elicit various effects on toxicity. Alcohol extract causes higher toxicity than aqueous extract.
Eubacterium and Bifidobacterium in the intestinal tract can hydrolyze saponins to produce toxic substances.
are fed with 25 mL/kg and 40 mL/kg ethanol and water extracts
of B. chinense DC. B. chinense DC and B. scorzonerifolium Willd
alcohol extracts also elicit higher toxicity than aqueous extracts.63
Influence of metabolites in vivo
Studies have proven that Eubacterium and Bifidobacterium in
the intestinal tract can hydrolyze saikosaponins to yield active
metabolites, which elicit overt toxicity.64
Pharmacokinetic studies in vivo have suggested that
saikosaponins can be digested and perform appropriate functions
only after these substances are modified by Enterobacteria in rats.
This finding indicates that metabolism in vivo influences the toxic
effects of saikosaponins.65
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The main toxic mechanism of Radix Bupleuri
The toxic effects of Radix Bupleuri in clinical applications
have been gradually observed; studies have shown that effective
components and toxic ingredients contain saikosaponins, and
the liver is the main organ affected by toxicity. Different dosages
of radix crude extracts can cause different degrees of liver
damage, which mainly increases total bilirubin, glutamic-pyruvic
transaminase, and glutamic-oxaloacetic transaminase in the blood.
This finding indicates that bilirubin metabolism is decreased and
hepatocyte membrane permeability is increased. Under an optical
microscope, the treated rats exhibit pathological changes, such as
hepatocyte nucleus pyknosis, edema, and eosinophilic changes,
whereas the mass and ratio of liver weight and body weight
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increase.66
Sun33 found that rats fed with saikosaponin for a long period
show changes in the liver function and hepatocyte organic lesion;
in addition, Sun33 found that the hepatic damage is related to
peroxidative damage mechanism. Saikosaponin D is the main
saikosaponin and active ingredient; this component elicits strong
liver toxicity in vitro, which involves apoptosis and changes in
cell membrane permeability; therefore, saikosaponin D causes cell
damage and necrosis.67
Huang68 intravenously administered 36.075 g/kg of total
saikosaponin extract in mice to observe the changes in P450
(CYP450), alanine aminotransferase (ALT), and aspartate
aminotransferase (AST) in a time-toxicity manner.69 The result
showed that CYP450, ALT, and AST levels increased 2 h after
administration, indicating that enzyme levels are correlated with
extract dosage; furthermore, liver tissue is significantly damaged
after total saikosaponins are administered, thereby inducing liver
cell edema, fat denaturation, and scattered punctiform necrosis at
varying degrees.
Hepatotoxicity induced by total saikosaponin extract is related
to drug metabolism in the liver. Oxygen radicals are generated
when CYP450 interacts with drugs, possibly causing toxic effects
by interacting with the cell membrane or other cell components.
This mechanism is consistent with oxidative damage induced by
total saikosaponins.
Different dosages of total saikosaponin extracts are administered
intragastrically in mice to observe Glutathione-S-transferase
(GST) and lactate dehydrogenase (LDH) levels in a time-toxicity
manner. The result indicated that acute liver injury may occur
when mice receive a single oral dose of total saikoponin in a dosedependent manner.70
GST is an enzyme present in many tissues and can eliminate
free radicals and lipid peroxide inside the body. Changes in LDH
concentrations can indicate liver damage. For instance, the cell
cycle slows down and cell proliferation decreases when LDH
increases; this finding is one of the sensitive indexes that indicate
abnormalities in liver energy metabolism;71 this result also suggests
that total saikosaponins can cause hepatotoxicity by affecting liver
energy metabolism.72,73
Zhang et al.74 studied hepatotoxity induced by refined
products of saikosaponins and discussed the relationship
between hepatotoxicity and hepatic fibrosis to provide the
basis of hepatotoxity mechanism.75-77 Zhang et al. conducted
a histopathological study and observed the changes in
hydroxyproline in the serum and liver tissue. Masson staining
shows that the pathological changes in the liver collagen fiber
is evident; these fibers are connected to one another via the
surrounding hepatic lobule, thereby disrupting normal lobular
architecture and forming a false lobule. Long-term administration
of high-dose refined products from saikosaponin can also cause
liver fibrosis in rats.
Future directions
Although saikosaponin is not listed as a toxic material in materia
medica during production, this substance produces side effects
in practical applications. Saikosaponins and volatile oil are toxic
6
components of Radix Bupleuri; these components also present
inhibitory effects. Different varieties, processing methods, and
extraction methods influence the toxic effects of Radix Bupleuri,
particularly saikosaponin. The main target organ is the liver, and
hepatic damage is related to peroxidative damage.
Saikosaponins are the active ingredients and the main
component causing toxicity; hence, comprehensive toxicity
studies and intensive evaluation of Radix Bupleuri can provide
evidence to establish the safe dosage range of toxic materials and
the minimum recommended dosage of active ingredients to ensure
safety and quality of Radix Bupleuri in clinical applications.
However, studies on the efficacy and toxicity of modern and
traditional Chinese medicine are limited because such studies are
directed to pharmacological activities, extraction and separation of
chemical components, toxicology, and other individual subjects.
The study of the relationship among these three subjects is limited.
For these reasons, international and modern assessment methods
should be developed to evaluate the toxicity of traditional Chinese
drugs.
Therefore, toxicity studies of Radix Bupleuri are in a primary
stage. Further studies should be conducted to investigate the toxic
effect of Radix Bupleuri on specific targets; monomer compositions,
toxicological dynamics, and toxicological mechanisms should also
be determined.
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