Diabetologia (2008) 51:216–226

DOI 10.1007/s00125-007-0886-7

REVIEW

The mechanisms of alloxan- and streptozotocin-induced

diabetes
S. Lenzen

Received: 12 September 2007 / Accepted: 8 October 2007 / Published online: 18 December 2007
# Springer-Verlag 2007

Abstract Alloxan and streptozotocin are toxic glucose Keywords Alkylation . Alloxan diabetes .
analogues that preferentially accumulate in pancreatic Cytotoxic glucose analogues . Pancreatic beta cell toxicity .
beta cells via the GLUT2 glucose transporter. In the Reactive oxygen species . Streptozotocin diabetes
presence of intracellular thiols, especially glutathione,
alloxan generates reactive oxygen species (ROS) in a Abbreviations
cyclic redox reaction with its reduction product, dialuric GSH glutathione
acid. Autoxidation of dialuric acid generates superoxide MNU N-methyl-N-nitrosourea
radicals, hydrogen peroxide and, in a final iron-catalysed NO nitric oxide
reaction step, hydroxyl radicals. These hydroxyl radicals PARP poly(ADP-ribose) polymerase
are ultimately responsible for the death of the beta cells, ROS reactive oxygen species
which have a particularly low antioxidative defence SOD superoxide dismutase
capacity, and the ensuing state of insulin-dependent
‘alloxan diabetes’. As a thiol reagent, alloxan also
selectively inhibits glucose-induced insulin secretion Introduction
through its ability to inhibit the beta cell glucose sensor
glucokinase. Following its uptake into the beta cells, Alloxan and streptozotocin are the most prominent diabe-
streptozotocin is split into its glucose and methylnitro- togenic chemicals in diabetes research. Both are cytotoxic
sourea moiety. Owing to its alkylating properties, the glucose analogues. Although their cytotoxicity is achieved
latter modifies biological macromolecules, fragments via different pathways, their mechanisms of beta cell
DNA and destroys the beta cells, causing a state of selective action are identical.
insulin-dependent diabetes. The targeting of mitochon- In 1838, Wöhler and Liebig [1] synthesised a pyrimidine
drial DNA, thereby impairing the signalling function of derivative, which they later called alloxan [2, 3]. In 1943,
beta cell mitochondrial metabolism, also explains how alloxan became of interest in diabetes research when Dunn
streptozotocin is able to inhibit glucose-induced insulin and McLetchie reported that it could induce diabetes in
secretion. animals [4] as a result of the specific necrosis of the
pancreatic beta cells [5–7]. The resulting insulinopenia
causes a state of experimental diabetes mellitus called
‘alloxan diabetes’ [4, 8, 9]. The reduction product of
alloxan, dialuric acid [1], has also been shown to be
diabetogenic in animals [10, 11], and to cause ultrastruc-
tural changes identical to those observed in response to
S. Lenzen (*)
Institute of Clinical Biochemistry, Hannover Medical School,
alloxan [6].
30623 Hannover, Germany Streptozotocin is an antimicrobial agent and has also
URL: http://www.mh-hannover.de/klinische_biochemie.html been used as a chemotherapeutic alkylating agent [12–14].
Diabetologia (2008) 51:216–226 217

In 1963, Rakieten et al. [15] reported that streptozotocin is injection, since streptozotocin does not inhibit glucoki-
diabetogenic. Again, this insulinopenia syndrome, called nase. Morphological alterations are minimal during this
‘streptozotocin diabetes’ [13], is caused by the specific phase.
necrosis of the pancreatic beta cells and streptozotocin has The second phase starts with an increase in the
been the agent of choice for the induction of diabetes blood glucose concentration, 1 h after administration of
mellitus in animals ever since [3, 16]. the toxins, and a decrease in plasma insulin. This first
After decades of research a unifying explanation for the hyperglycaemic phase, which usually lasts 2–4 h, is
selective toxicity of these two most prominent diabetogenic caused by inhibition of insulin secretion leading to
agents [2, 17–22] can be provided. Since an understanding hypoinsulinaemia. During this phase the beta cells show
of the chemical reactivity of these compounds is crucial for the following morphological characteristics: intracellular
understanding their diabetogenicity, this review will pro- vacuolisation, dilation of the rough endoplasmic retic-
vide an integrated view of their chemical properties and ulum, decreased Golgi area, reduced secretory granules
biological effects. and insulin content, and swollen mitochondria.
The third phase, again a hypoglycaemic phase,
typically occurs 4–8 h after the injection of the toxins
and lasts several hours. It may be so severe that it
Alloxan diabetes and streptozotocin diabetes causes convulsions, and may even be fatal without
glucose administration, in particular when liver glyco-
Figure 1 shows a schematic diagram of the tetraphasic gen stores are depleted through starvation. This severe
and triphasic blood glucose responses induced by alloxan transitional hypoglycaemia is produced by the flooding
and streptozotocin, respectively, when injected [22]. The of the circulation with insulin as a result of toxin-
responses are accompanied by corresponding inverse induced secretory granule and cell membrane rupture.
changes in plasma insulin and sequential ultrastructural Pancreatectomy prevents this phase. In addition to the
changes resulting in necrotic beta cell death. morphological changes seen in the first phase, the beta
A first transient hypoglycaemic phase of up to cell nuclei are pyknotic and show no TUNEL-positive
30 min starts within minutes of alloxan injection. This staining; these changes are irreversible.
short-lived hypoglycaemic response is the result of a The fourth phase is the permanent diabetic hyper-
transient stimulation of insulin secretion, as documented glycaemic phase. Morphologically, complete degranula-
by an increase in the plasma insulin concentration. The tion and loss of beta cell integrity is seen within 12–
underlying mechanism is a temporarily reduced con- 48 h. Non-beta cells remain intact, demonstrating the
sumption and increased availability of ATP caused by beta cell-selective character of the toxic action. Cell
blockade of glucose phosphorylation through glucoki- debris originating from the dying beta cells is removed
nase inhibition. This initial transient hypoglycaemic by non-activated scavenger macrophages.
phase is not observed in response to streptozotocin Thus, injections of alloxan and streptozotocin prin-
cipally induce the same blood glucose and plasma
insulin responses and cause an insulin-dependent type
1-like diabetes syndrome. All of the described morpho-
logical features of beta cell destruction are characteris-
tic of necrotic cell death [22]. This mechanism is clearly
at variance with that which underlies autoimmune type 1
diabetes, both in humans and rodent models of the
disease, where beta cell demise is the result of apoptotic
cell death without leakage of insulin from ruptured
secretory granules [22].

Alloxan: mechanism of action

Fig. 1 Phasic blood glucose response to a diabetogenic dose of

Alloxan has two distinct pathological effects: it selec-
alloxan (tetraphasic; I–IV) or streptozotocin (triphasic; the first phase tively inhibits glucose-induced insulin secretion through
does not develop in the case of streptozotocin; II–IV) specific inhibition of glucokinase, the glucose sensor of
218 Diabetologia (2008) 51:216–226

the beta cell, and it causes a state of insulin-dependent Beta cell selectivity of alloxan
diabetes through its ability to induce ROS formation,
resulting in the selective necrosis of beta cells. These Alloxan is a very unstable chemical compound [23]
two effects can be assigned to the specific chemical with a molecular shape resembling glucose (Fig. 2) [24,
properties of alloxan, the common denominator being 25]. Both alloxan and glucose are hydrophilic and do not
selective cellular uptake and accumulation of alloxan penetrate the lipid bilayer of the plasma membrane. The
by the beta cell. alloxan molecule is structurally so similar to glucose that

Comparison of the chemical properties of alloxan and streptozotocin

Alloxan Streptozotocin
Chemical name 2,4,5,6-Tetraoxypyrimidine; 2-Deoxy-2-
2,4,5,6-pyrimidinetetrone ([(methylnitrosoamino)carbonyl]amino)-
D-glucopyranose
Chemical structure Oxygenated pyrimidine derivative; Cytotoxic methylnitrosourea moiety (N-
barbituric acid derivative (5- methyl-N-nitrosourea) attached to the
ketobarbituric acid) glucose (2-deoxyglucose) molecule;
glucosamine derivative
Chemical properties Very hydrophilic, beta cell-toxic Hydrophilic, beta cell-toxic glucose
glucose analogue (partition analogue
coefficient –1.8); weak acid
Chemically unstable (half-life of Relatively stable at pH 7.4 and 37o C (at
1.5 min at pH 7.4 and 37o C, least for up to 1 h)b
decomposing to alloxanic acid);
a
Stable at acid pH
Chemical reactivities Thiol reagent that is reduced to DNA alkylating agent
dialuric acid in the presence of
GSH and other thiols
A protoxin; intracellular Protein alkylating agent
metabolism of this xenobiotic
generates toxic ROS through
redox cycling with dialuric acid
over a long time period (>1 h)
‘Compound 305’, a non-toxic NO donor
alloxan-GSH adduct of unknown
structure with a characteristic
absorbance at a wavelength of 305
nm; a small amount is formed
during each redox cycle
Mode of toxicity Generation of ROS DNA alkylation
a
For experimentation, concentrated stock solutions in 0.01 mol/l HCl, kept on ice, should be used and
added to the test medium just prior to the start of the experiment to obtain the final concentration. For
injection, the stock solution should be diluted with ice-cold saline (0.9% NaCl) immediately prior to
injection.
b
For in vitro experimentation, concentrated stock solutions in 0.01 mol/l HCl, kept on ice, should be
used and added to the test medium just prior to the start of the experiment to obtain the final
concentration. For injection, a stable solution in citrate buffer (pH 4.5) is most suitable.
Diabetologia (2008) 51:216–226 219

Fig. 2 Chemical formulas of

alloxan, dialuric acid,
butylalloxan, streptozotocin
and methylnitrosourea

the GLUT2 glucose transporter in the beta cell plasma [2, 40], with a half maximal inhibitory concentration in the
membrane accepts this glucomimetic and transports it into the 1–10 μmol/l range. At higher concentrations, alloxan can
cytosol [25, 26]. Alloxan does not inhibit the function of the inhibit many functionally important enzymes, as well as
transporter [27], and can therefore selectively enter beta cells other proteins and cellular functions [22, 41].
in an unrestricted manner [28–30]. It is therefore not toxic to Inhibition of glucokinase reduces glucose oxidation and
insulin-producing cells that do not express this transporter [27, ATP generation [42], thereby suppressing the ATP signal
31]. The half-life of alloxan is short [23, 32]; in aqueous that triggers insulin secretion [2]. Inhibition of glucokinase
solution it spontaneously decomposes into non-diabetogenic is achieved within 1 min of exposure to alloxan.
alloxanic acid within minutes [23]. Because of this, it must be The inhibition of glucose-induced insulin secretion is
taken up and accumulated quickly in the beta cell [28], and is preceded by a very transient (1–2 min) stimulation of insulin
therefore ineffective when blood flow to the pancreas is secretion immediately after exposure to alloxan [43]. This ef-
interrupted for the first few minutes after alloxan injection fect can be explained by an initial reduction of ATP consump-
[33–35]. N-substituted alloxan derivatives with a long carbon tion resulting from the blockade of glucose phosphorylation
side chain, such as butylalloxan (Fig. 2), differ chemically by glucokinase [44], which produces a transient increase in
from alloxan in that they are lipophilic [36]. Butylalloxan acts ATP in the beta cell and triggers a transient release of insulin.
in a similar manner to alloxan and preferentially damages beta The inhibition of insulin secretion after exposure to alloxan
cells [6, 11]. But since derivatives such as butylalloxan are [43, 45, 46] is restricted to that induced by glucose and its
lipophilic they can also penetrate plasma membranes that do epimer, mannose, both of which induce insulin secretion
not express the GLUT2 transporter [27]. Nephrotoxicity is a through interaction with glucokinase [44]. Insulin biosynthesis
dominating feature of the toxicity of lipophilic derivatives is also inhibited by alloxan [47, 48], most likely through the
after systemic administration [11]. This nephrotoxicity is so same mechanism. The insulin secretory response to other
severe that it causes fatal renal failure in the animals before nutrient secretagogues, such as 2-ketoisocaproic acid and
diabetes can develop [11]. The susceptibility of the kidney to leucine, or non-nutrient secretagogues, such as the sulfonyl-
the toxic action of these lipophilic alloxan derivatives is the urea drug tolbutamide, remains intact initially because it is not
result of their preferential accumulation in the tubular cells of mediated via glucokinase [49], but is lost after a delay of up to
the kidney, which, like the beta cells, express the GLUT2 1 h [50] as a consequence of the gradual deterioration of beta
glucose transporter. cell function.
Thiols such as the tripeptide glutathione (GSH), cysteine and
Glucokinase inhibition dithiothreitol protect glucokinase against alloxan inhibition
because they reduce alloxan to dialuric acid, which is not thiol
Selective inhibition of glucose-induced insulin secretion is the reactive [23, 51, 52]. However, only dithiols such as
major pathophysiological effect of the thiol group reactivity of dithiothreitol [51, 52] can readily reverse alloxan-induced
alloxan [37–39]. Alloxan has a central 5-carbonyl group that glucokinase inhibition. They achieve this by reducing func-
reacts very avidly with thiol groups. Glucokinase (hexoki- tionally essential cysteine residues of the glucokinase enzyme
nase IV) is the most sensitive thiol enzyme in the beta cell [40], which are oxidised through alloxan action [51, 52].
220 Diabetologia (2008) 51:216–226

Likewise, glucose protects against alloxan-induced inhibi- redox cycling reactions between alloxan and dialuric acid,
tion of glucose-induced insulin secretion because its binding and protective actions of cytoprotective enzymes’ (reactions
to the sugar-binding site of glucokinase prevents the oxidation i–ii). In the beta cells the toxic action of alloxan is initiated
of the functionally essential thiol groups. The non-metabolis- by free radicals formed in this redox reaction [53–57].
able seven carbon sugar mannoheptulose protects glucokinase Autoxidation of dialuric acid generates superoxide radicals
through the same mechanism, but this alone is not sufficient to (iii–iv) and hydrogen peroxide (iii–iv), and in the Fenton
prevent alloxan-induced inhibition of insulin secretion. The reaction (v), in the presence of a suitable metal catalyst
glucose analogue 3-O-methylglucose, which is not a sub- (typically iron) (vi), hydroxyl radicals (v–vii). The autox-
strate of glucokinase, does however prevent inhibition. It idation of dialuric acid involves the intermediate formation
does this through competitive blockade of alloxan uptake of the alloxan radical (i–iv) [54–56].
into the beta cell via the GLUT2 glucose transporter. The reduction of alloxan to dialuric acid in the cell
Thus, the inhibition of glucose-induced insulin secretion requires the presence of a suitable thiol, typically the
by alloxan is the result of the exquisite thiol reactivity of tripeptide glutathione (GSH) to generate the redox cycling
the glucose sensor glucokinase. partner, dialuric acid, and oxidised glutathione (viii) [56,
58]. The triketone structure of alloxan is vitally important
Beta cell toxicity and diabetogenicity of alloxan for this two-step reaction with glutathione [59], which
generates the alloxan radical as an intermediate product
Alloxan can generate reactive oxygen species (ROS) in a (ix–x). Other thiols such as cysteine, which are present at
cyclic reaction with its reduction product, dialuric acid lower concentrations in the cell, dithiols and ascorbic acid
(Fig. 3) [53–56], as depicted in the text box ‘Chemical are also suitable reducing agents and may therefore
contribute to alloxan reduction [60]. Alloxan can also
generate ROS by reacting with thiol groups on proteins
Chemical redox cycling reactions between such as enzymes [52, 61] and albumin [62]. During each
alloxan and dialuric acid, and protective typical redox cycle a small amount of ‘Compound 305’, an
actions of cytoprotective enzymes alloxan–GSH adduct [23, 32, 56, 63], is formed. The
. . intracellular concentration of Compound 305 increases in
(i) AH 2 + O 2 AH + O 2− + H + a time-dependent manner, which gradually decreases the
. .− amount of reduced GSH available in the cell for redox
(ii) AH + O 2 A + O2 + H + cycling, thus producing a lower pro-oxidative ratio
.− . between alloxan and GSH, rather than a higher antiox-
(iii) AH 2 + O 2 + H + AH + H 2 O 2
idative ratio [54–56].
. .−
(iv) AH + O 2 + H + A + H 2O 2 Paradoxically the thiols cysteine and GSH have long
been reported to protect rats against the development of
.
(v) H 2 O 2 + e − OH + OH − alloxan diabetes when injected together with alloxan [64,
. 65]. This observation can now be explained in light of the
(vi) Fe 2+ + H 2 O 2 Fe 3+ + OH + OH − established molecular mechanism of alloxan action. When
.− metal . concentrations of reducing agents in the blood stream or in
(vii) Net : O 2 + H 2 O 2 O 2 + OH + OH −
catalyst the extracellular space are significantly increased through
(viii) A + 2GSH AH 2 + GSSG injection of a thiol, more alloxan is reduced extracellularly
. . so that less is available for intracellular accumulation.
(ix) A + GSH AH + GS Normally the capacity for alloxan reduction, redox cycling
. . and the generation of ROS in the circulation [62] is not
(x) AH + GSH AH 2 + GS sufficient to prevent the alloxan molecule from reaching
.− and entering the beta cell.
(xi) 2H + + 2O 2 SOD
H 2O 2 + O 2
The major oxidation pathway of dialuric acid, a chain
(xii) 2H 2O 2 Catalase
O 2 + 2H 2O reaction dependent upon superoxide radicals, is inhibited by
superoxide dismutase (SOD; xi). In the presence of SOD,
(xiii) 2GSH + H 2O 2 GPx
2H 2O + GSSG an autocatalytic process involving the interaction between
. dialuric acid and alloxan becomes important [54], while in
A, alloxan; AH , alloxan radical; AH2, dialuric acid;
. the presence of a transition metal, a third oxidation
GPx, glutathione peroxidase; GS , glutathione radical;
. mechanism, dependent upon hydrogen peroxide, has been
GSSG, oxidised glutathione; OH , hydroxyl radical;
. identified [54]. This latter step is inhibited by the hydrogen
O 2− , superoxide radical
peroxide inactivating enzyme catalase [54] (xii; text box:
Diabetologia (2008) 51:216–226 221

Several lines of evidence point to hydroxyl radicals as the

principal culprit.
First, the hydrogen peroxide-inactivating enzyme cata-
lase provides significantly better protection against the
toxic effects of alloxan and dialuric acid on insulin-
producing cells than the superoxide radical-inactivating
enzyme SOD, though catalase does not prevent redox
cycling and therefore superoxide radical formation [54, 56].
Second, since intracellular GSH concentrations are in the
same millimolar concentration range in insulin-producing
cells as in liver cells, and since both cell types express the
GLUT2 glucose transporter, a difference in the intracellular
GSH concentration cannot be responsible for the much
Fig. 3 Redox cycling reactions between alloxan and dialuric acid. A,
greater susceptibility in vivo of insulin-producing cells
alloxan; AH , alloxan radical; AH2, dialuric acid; GS , glutathione compared with liver cells to the toxicity of alloxan [17].
radical; GSSG, oxidised glutathione; OH , hydroxyl radical; O2 , However, liver cells are much better endowed with the
superoxide radical hydrogen peroxide-inactivating enzyme catalase compared
with insulin-producing cells [66, 67]. When the low
Chemical redox cycling reactions between alloxan and intracellular levels of catalase are raised in insulin-producing
dialuric acid, and protective actions of cytoprotective cells through overexpression of the gene for this enzyme,
enzymes). The other hydrogen peroxide inactivating en- these cells are protected equally well [60].
zyme, glutathione peroxidase, can principally act in a Third, a multitude of metal chelators, iron chelators in
similar manner. But this enzyme requires GSH, which is particular, and hydroxyl scavengers have been tested for
oxidised in this reaction (xiii). their protective action both in vitro and in vivo, but the
When kept in the oxidised form, alloxan does not results obtained have not been unequivocal [18, 22, 66, 68–
generate ROS [60]. Thus, alloxan is not cytotoxic in the 70]. Given the extremely short half-life and the extraordi-
absence of thiols such as GSH or when restricted to the nary chemical reactivity of the hydroxyl radical, it is not
extracellular space [60]. Thiols in the plasma membrane, surprising that complete protection against alloxan toxicity
with which alloxan could interact and generate ROS in a cannot be achieved. Hydroxyl radicals interact with
redox cycle, are apparently not present or not accessible to biological targets before they can be inactivated through
a sufficient extent to allow the generation of ROS and hydroxyl radical scavengers. In the same way, it is not
damage the cells [60]. surprising that it has been extremely difficult to efficiently
The reduction product dialuric acid is also not toxic suppress metal-catalysed hydroxyl radical formation
when kept in the reduced form [54, 56, 60]. However, in through chelators [69–72], since it is crucial that metal
contrast to alloxan, dialuric acid autoxidises spontaneously chelation occurs before hydroxyl radical formation is
in the presence of O2, thus generating cytotoxic ROS in the initiated. Owing to the chemical properties of the scav-
absence of thiols [54, 55]. As a result of subsequent redox engers and chelators, this cannot be completely achieved
cycling it can also induce diabetes [10, 11] and cause beta unless experimental conditions are optimised. In such a
cell lesions as alloxan [6]. setting the toxicity of alloxan and dialuric acid to insulin-
The antioxidative enzyme SOD has a cytoprotective producing cells in vitro is suppressed by the iron chelator
effect against both alloxan and dialuric acid in the presence desferrioxamine [60], which prevents the generation of the
of GSH by virtue of its ability to scavenge superoxide very toxic hydroxyl radicals in the iron-catalysed Fenton
radicals, which are generated in the O2-dependent chain reaction [54, 56] (Fig. 3).
reaction between dialuric acid and alloxan. The resultant Taken together, these data provide convincing evidence
suppression of dialuric acid autoxidation prevents the that the hydroxyl radical [56, 60, 73], rather than the
generation of further ROS [54, 56], although increasing superoxide radical, is the ultimate toxic ROS species, and
the concentrations of the toxins can reinstate the toxic that its formation is prevented by the destruction of
action of both compounds. This is due to an autocatalytic hydrogen peroxide by catalase [54, 56].
reaction between the oxidised and reduced pyrimidine, Optimal protection against the cytotoxic action of
which generates ROS even when the chain reaction is alloxan and dialuric acid is provided only by a combination
suppressed by SOD [54, 56]. of SOD plus catalase, which completely prevents redox
Apparently, the superoxide radical is not the species cycling between alloxan and dialuric acid, and thus the
responsible for the cytotoxicity of alloxan and dialuric acid. generation of all ROS species in this reaction pathway.
222 Diabetologia (2008) 51:216–226

Glucose also provides complete protection against all trosourea moiety [79–83], especially at the O6 position of
toxic effects of alloxan both in vivo and in vitro [2]. This guanine [22]. The transfer of the methyl group from
universal protection is achieved through the prevention of streptozotocin to the DNA molecule causes damage, which
glucokinase inhibition and the preservation of the antiox- along a defined chain of events [84], results in the
idative defence mechanisms of the beta cell [22]. The non- fragmentation of the DNA [85]. Protein glycosylation may
metabolisable glucose analogue 3-O-methylglucose also be an additional damaging factor [41]. In the attempt to repair
provides protection, but does this exclusively through the DNA, poly(ADP-ribose) polymerase (PARP) is overstimu-
prevention of alloxan uptake into the beta cell via the lated. This diminishes cellular NAD+, and subsequently ATP,
GLUT2 glucose transporter [22]. stores [82, 85–87]. The depletion of the cellular energy stores
Thus, it can be concluded that the pancreatic beta cell ultimately results in beta cell necrosis. Although streptozotocin
toxicity and the resultant diabetogenicity of alloxan are due also methylates proteins [80, 81], DNA methylation is
to redox cycling and the generation of toxic ROS. ultimately responsible for beta cell death, but it is likely that
protein methylation contributes to the functional defects of the
beta cells after exposure to streptozotocin.
Streptozotocin: mechanism of action Inhibitors of poly ADP-ribosylation suppress the process
of DNA methylation. Thus, injection of nicotinamide and
Streptozotocin (Fig. 2) inhibits insulin secretion and causes other PARP inhibitors in parallel with, or prior to the
a state of insulin-dependent diabetes mellitus. Both effects administration of streptozotocin is well known to protect
can be attributed to its specific chemical properties, namely beta cells against the toxic action of streptozotocin and to
its alkylating potency (see text box: Comparison of the prevent the development of a diabetic state [13]. Also, mice
chemical properties of alloxan and streptozotocin). As with deficient in PARP are resistant to beta cell death mediated
alloxan, its beta cell specificity is mainly the result of by streptozotocin, in spite of DNA fragmentation. The
selective cellular uptake and accumulation. absence of PARP prevents the depletion of the cofactor
NAD+ and the subsequent loss of ATP [84, 88–90] and thus
cell death.
Beta cell selectivity of streptozotocin The role of alkylation in beta cell damage has also been
examined by the use of ethylating agents, which are less
Streptozotocin is a nitrosourea analogue in which the toxic than their methylating counterparts, on account of O6-
N-methyl-N-nitrosourea (MNU) moiety (Fig. 2) is linked ethylguanine being less toxic than O6-methylguanine [91].
to the carbon-2 of a hexose. The toxic action of The fact that N-ethyl-N-nitrosourea and ethyl methane-
streptozotocin and chemically related alkylating compounds sulphonate are significantly less toxic to insulin-producing
requires their uptake into the cells. Nitrosoureas are usually
lipophilic and tissue uptake through the plasma membrane
Beta cell-toxic
is rapid; however, as a result of the hexose substitution, glucose analogues Alloxan Streptozotocin
streptozotocin is less lipophilic. Streptozotocin is selective-
ly accumulated in pancreatic beta cells via the low-affinity
Beta cell-selective Selective beta cell uptake via the
GLUT2 glucose transporter in the plasma membrane [74, action
GLUT2 glucose transporter
75]. Thus, insulin-producing cells that do not express this
glucose transporter are resistant to streptozotocin [76–78].
This observation also explains the greater toxicity of Mechanism of Beta cell toxicity Beta cell toxicity
streptozotocin compared with N-methyl-N-nitrosourea in
beta cell death
through ROS through alkylation
cells that express GLUT2, even though both substances
alkylate DNA to a similar extent [77, 79, 80]. The Beta cell death
Mode of
importance of the GLUT2 glucose transporter in this beta cell death
through necrosis
process is also shown by the observation that streptozotocin
damages other organs expressing this transporter, particu- Insulin-dependent
Beta cell death result
larly kidney and liver [17, 19]. diabetes mellitus

Chemical diabetes Alloxan Streptozotocin

Beta cell toxicity of streptozotocin
diabetes diabetes
Fig. 4 Schematic representation of the toxic effects of the glucose
It is generally assumed that the toxicity of streptozotocin is analogues alloxan and streptozotocin in beta cells, which produce
dependent upon the DNA alkylating activity of its methylni- chemical diabetes
Diabetologia (2008) 51:216–226 223

cells than MNU and methyl methanesulphonate [77, 91] exposure, while long-term inhibition of insulin secretion
has been taken as support for the notion that in insulin- 6 days after streptozotocin exposure was not counteracted
producing cells, as in other cell types, the mechanism of by nicotinamide [100].
toxic action is due to alkylation, with methylation of DNA
bases being more toxic than ethylation [22, 81].
An alternative hypothesis proposes that part of the Conclusion
diabetogenic effect of streptozotocin may relate not to its
alkylating ability but to its potential to act as an intracellular Both alloxan and streptozotocin induce insulin deficiency.
nitric oxide (NO) donor [92]. Both streptozotocin and While the mechanisms of beta cell-selective action through
MNU contain a nitroso group (Fig. 2) and can liberate NO. uptake via the GLUT2 glucose transporter and beta cell
In fact, streptozotocin has been shown to increase the death via necrosis are identical, ROS in the case of alloxan
activity of guanylyl cyclase and the formation of cGMP, and DNA alkylation in the case of streptozotocin mediate
which are characteristic effects of NO. However, the the toxic action of these glucose analogues (Fig. 4). This
alkylating agent methyl methanesulphonate is the most also explains why a lack of significant expression of this
toxic compound, though unlike MNU, it is not a NO donor glucose transporter isoform, such as in human beta cells,
[91], indicating that NO is not an indispensable prerequisite provides insensitivity to the toxins [22, 101]. On the other
for the toxic action of the family of alkylating agents that hand, expression of this glucose transporter in other organs
streptozotocin belongs to. like in tubular cells and hepatocytes explains why the
Finally, some minor generation of ROS, including toxins can cause damage to kidney and liver [22].
superoxide and hydroxyl radicals originating from hydro- Due to its chemical properties, in particular the greater
gen peroxide dismutation during hypoxanthine metabolism stability (see text box: Comparison of the chemical
[93], may accompany the effect of streptozotocin and properties of alloxan and streptozotocin), streptozotocin is
accelerate the process of beta cell destruction but ROS do the agent of choice for reproducible induction of a diabetic
not play a crucial role [22]. metabolic state in experimental animals. Alloxan, on the
other hand, as a model compound of ROS mediated beta
cell toxicity, is the agent with the greater impact upon the
Inhibition of insulin secretion by streptozotocin understanding of ROS mediated mechanisms of beta cell
death in type 1 and type 2 diabetes mellitus.
The effects of streptozotocin on glucose and insulin homeo-
stasis reflect the toxin-induced abnormalities in beta cell Acknowledgements The author is most grateful to Frits Holleman
for excellent editorial support.
function. Initially, insulin biosynthesis, glucose-induced insu-
lin secretion and glucose metabolism (both glucose oxidation Duality of interest The author states no duality of interest.
and oxygen consumption) are all affected [93–95]. On the
other hand, streptozotocin has no immediate, direct inhibito-
ry effect upon glucose transport [77] or upon glucose
phosphorylation by glucokinase [39]. However, at later
stages of functional beta cell impairment, deficiencies in References
terms of gene expression and protein production lead to the
1. Wöhler F, Liebig J (1838) Untersuchungen über die Natur der
deterioration of both glucose transport and metabolism [96].
Harnsäure. [Investigations on the nature of uric acid]. Ann
Even before the negative effects of mitochondrial DNA Pharm 26:241–340 (article in German)
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