The British HomoeopathicJournal

July 1988. Vol. 77. pp. 147-151

Pancreatic beta-cell regeneration

a novel anti-diabetic action o f Cephalendra indica m o t h e r tincture
D. P. R A S T O G I * , A. C. S A X E N A t , SUNIL K U M A R $

Abstract

Cephalendra indica 0 (41% v/v alcoholic extract of the wild variety of Cephalendra
indica Naud.), on regular administration in doses ranging from 25 ~tml to 75 Ixml/100 g of
body weight (gbw) by the oral or intraperitoneal (ip) route produced a significant fall in
blood sugar level in alloxan-induced diabetic rats. Biochemical studies showed stabilization of blood sugar level in 70% of cases at fourteen to twenty days after withdrawal
of the drug. Histopathological studies revealed regeneration of pancreatic [3 cells. The
hypothesis is that the drug acts through the hypothalamo-hypophysial-pancreatic axis,
producing selective regeneration of 13ceils. The drug may indirectly release inhibitory
factors from hypothalamic neurons, inhibiting the secretion of growth hormone and
triggering insulin secretion from [3cells. The therapeutic action of the drug on pancreatic
[3 cells and lack of acute and subacute toxicity may open up new prospects in the
treatment of diabetes mellitus.

(C4H2N204H20,M.W.160.09, purity 98.5 %). In

the present investigation, an attempt has been
made to locate the mode of action of the drug
with particular emphasis on [3 cell regeneration
and its neuroendocrinological control.

Introduction

Treatment of diabetes has been reviewed by

many researchers. 1-4 The causative factor of
diabetes has attracted considerable investigation
over the centuries and still remains uncertain.
Polyuria and wasting are the dominant clinical
features: ' A melting down of the flesh and limbs
into urine' (Aretaeus, 2nd century A D ) , and it
was long thought to be due to kidney disease.
Recently, extensive data have accumulated to
establish that renal disease is secondary to
glycosuria. ~6 Agrawal and Bapat 7 on the basis of
their piqure experiments believe that diabetes is
due to disease of the central nervous system.
There are undoubtedly many different causes of
diabetes. Advances in genetics, the concept of
neuroendocrinology, understanding of major
histocompatibility complex and developments in
immunology have considerably advanced our
understanding of the nature of the disease.
We have experimented with mother tincture of Cephalendra indica in diabetes
mellitus experimentally produced by alloxan

Materials and methods

*D. P. Rastogi, Director, Central Council for Research in

Hom0eoapthy (Min. of Health & Family Welfare, Govt. of
India).
?A. C. Saxena,ProjectDirector,HomcepathicDrug Research
Institute, SectorB-1433, Indira Nagar, LUCKNOW-226016.
:~SunilKumar, ResearchOfficer(P), HDRI.

One hundred and eighty albino rats of either sex

weighing 240+12 g were selected after acclimatizing to standard laboratory conditions for
fifteen days. Water was allowed ad-libitum.
Photo-period L/D (10 light hours/14 dark hours)
was also maintained. The acclimatized animals
were subjected to quantitative analysis of blood
sugar by the Folin and Wu method by taking a
0.5 ml blood sample from the tail vein or through
cardiac puncture and measuring absorbance at
620 nm wave length in a Beckman Model 35
spectrophotometer.
Diabetes mellitus was induced by intraperitoneal injections. Three doses of 10 to 12
rag/100 gbw, at intervals of seven days, of
alloxan dissolved in distilled water, were administered after twelve hours fasting. Blood sugar
estimations were done to confirm the establishment of diabetes mellitus. The diabetized animals were divided into six groups for in-vivo and
in-vitro studiesl The short-term experiment was
conducted over twenty one days and the long-

147

148

British Homoeopathic Journal

Figure 1: Degenerative changes with prominent beta cell

necrosis after alloxan treatment. Gomori's aldehyde fuchsin
x 500.

Figure 2: Cephalendro indica 0 treated after 30th day

(25 uml/100 gbw), with prominent 13cell restoration. Gomori's
aldehyde fuchsin

Figure 3: Cephalendra indica r treated after 30th day

(75 uml/100 gbw) with large number of [3 cells in islet of
Langerhans. Ehrlich's haematoxylin and Eosin

Figure 4: Cephalendra indica ~ treated after 30th day

(25 ~tml/100 gbw) S.S. of brain, passing through hypothalamus. Non-secretory activity with hypotrophic neurons and
significant GH-IF. Gomori's aldehyde fuchsin x500.

Figure 5: 41% v/v alcohol fed control after 30th day. S.S. of
brain, passing through hypothalamus. Perceptibile secretory
activity with prominent hypertrophied neurons and indicating
GH-RF. Gomori's aldehyde fuchsin x500.

149

Volume 77, Number3, July 1988

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term experiment over forty five days. The paral

lel studies were conducted with tolbutamide
(Rastinon), glibenclamide (Daonil) and insulin.
A market sample of Cephalendra indica ~ was
studied in addition to a tincture prepared in the
Homceopathic
Pharmacopoeia
Laboratory,
Ghaziabad, and the therapeutic efficacy of the
drug compared with the 41% v/v alcohol control,
since the ~ is prepared in 41% v/v alcohol as
specified in the Homceopathic Pharmacopoeia of
India. Both the oral and the i,p. route of drug
administration were attempted. The normal
control group was given 0.9% w/v physiological
saline. The diabetized rats were divided into
groups of ten as follows:
The long4erm experiment was conducted for
forty-five days, but the drug was only administered for the first thirty days. After that, drug
administration was stopped and the animals
assessed for blood sugar stabilization.
The blood sugar estimation was done at
twelve hours fasting and two hours postprandial
on the first, fifth, seventh, fourteenth, twenty
first and thirtieth days. The histopathological
studies were conducted on a 25% sample of the
experimental animals. The brain, pituitary
gland, pancreas, liver, kidney etc., were isolated
and processed for histopathological studies.
These organs/tissues were isolated by decapita
tion of animals, after which the entire retroperitoneal fat containing pancreatic tissue was
dissected out and fixed in freshly prepared
Bouin's fluid. The tissues were cut into two to
four #m thick sections and stained in haematoxylin eosin and Gomori aldehyde fuchsin stain
with aqueous light green 0.6 g + chromotroph2 R 0.5 g + orange G 1.0 g + glacial acetic acid
1 ml/100ml of water as counter stain. The [3 cells
per islet area in cross section (mm x 350) were
counted under a light microscope.
Results

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Experimental studies revealed that regular

administration of Cephalendra indica 0 for
thirty days in micro doses ranging from 25 gml.
to 75 ,aml./100 gbw produced a slow and steady
fall in blood sugar level, and hypoglyc~emic
activity came into prominence after the tenth to
fifteenth day of treatment. The normal control
and alcohol fed control groups continued to be
hyperglyca~mic (Table 1.)
The beta cell counts and blood sugar levels in
various groups of alloxanized rats are given in
Table 2. It is evident from the table that alloxan
brings about necrosis of ~3 cells and markedly

British Homceopathic Journal

150
TABLE 2

Influence of Cephalendra indica ~ on pancreatic beta cells and blood sugar levels in alloxanized rats
(Mean_+S.E.)
Cephalendra indica r administration on
30th day in per 100 gbw

Blood sugar
level/f3 cell
counts

Normal
control
0.9% w/v

Control
41% v/v
alcohol

25 gmI

50 gml

75 gml

0.1 gml

Blood sugar
level (fasting
in mg/dil.)

255-+3.2

266_+20

80+2.0

100_+1.8

90-+2.7

115+3.2

B~

C~

D~

E~

Fj

NS*
A~:C~ <0.001
A~:Dt <0.001
A]:E] <0.001
AI:FI <0.001

Beta cell
counts per islet
area in cross
section
( m m 2 X 350)

20

30+2.8

32+5.0

D~

E2

At

A2

18_+3.2

B2

36

C2

26

F,

AI:B 1

A2:B2 NS*
A2:C2 <0.001
A2:D2 <0.001
Ai:E 2 <0.001
A 2 : F 2 <0.001

*NS = Not significant. All statistical values calculated by Student t test.

TABLE3 Hypoglycaemic activities of Cephalendra indica r Stabilization of blood sugar level after withdrawal of drug
Drug/vehicles

Route of
admn.

Days at which blood

sugar level comes to
normal

Effective doses Duration of stabilization Percentage of

per 100 gbw
of blood sugar level stabilization after
withdrawal of drug ED 50 & ED 100
mean value
analysis

41% v/v alcohol

control

oral/i.p.

Exhibited
None effective
hyperglycaemic action

Cephalendra
indica ~

oraI/i.p.

Attained normal level

usually after 15 days of
regular drug admn. at a
dose level of 25 gml to
75 gml/100 gbw
depending on blood
sugar level

Never attained normal

level

Effective does are 14 to 20 days (in 30 days

25 gml to 75 gml, regular admn. of drug)
with 25 gml most No toxicity (30 days
effective
regular frug admn.)

Nil
70%
70%

'\

reduces the n u m b e r along with an increase in

blood sugar level (Fig. 1). T h e Cephalendra
indica ~ t r e a t e d animals exhibited b l o o d sugar
level and 13 cell counts within n o r m a l r a n g e at a
dose level ranging from 25 gml to 75 Bxml/100
gbw (Figs. 2 & 3).
Histological e x a m i n a t i o n of the brain
revealed certain h y p o t h a l a m i c n e u r o n s h a v i n g
specific structural peculiarities and containing
secretory material and p r o m i n e n t l y n u c l e a t e d
with bipolar axons. T h e s e structures are identified by special t e c h n i q u e s of tinctorial affinities
( d e v e l o p e d in o u r L a b o r a t o r y ) . T h e s e n e u r o n s
are juxtasomal with b l o o d vessels and responsible for the secretion of releasing factors (RF)
or inhibitory factors (IF) for growth h o r m o n e s
( G H ) . Excessive secretion of G H p r o b a b l y
inhibits insulin secretion and p r o d u c e s d i a b e t e s
mellitus by d a m a g i n g the [3 cells. H o w e v e r , in
the p r e s e n t e x p e r i m e n t it was n o t e d that the
drug h a d b r o u g h t a b o u t h y p o t r o p h y in h y p o t h -

alamic n e u r o n s (Fig. 4) and p r o b a b l y released

the IF which in t u r n i n h i b i t e d growth h o r m o n e
secretion from the a d e n o h y p o p h y s i s . This m a y
have resulted in the r e s t o r a t i o n of 13 cells counts'
a n d r e g e n e r a t i o n , w h e r e a s the control groups
h a d s h o w n h y p e r t r o p h y and d e g r a n u l a t i o n in
h y p o t h a l a m i c n e u r o n s (Fig. 5).
Table 3 shows the f r e q u e n c y of b l o o d sugar
level stabilization after withdrawal of the drug.
It was o b s e r v a b l e that the b l o o d sugar level was
stabilized for f o u r t e e n to twenty days in 70% of
cases in the Cephalendra indica ~ t r e a t e d group,
w h e n c o m p a r e d to control animals.
T h e parallel studies with t o l b u t a m i d e , glibe n c l a m i d e and insulin c o n d u c t e d u n d e r identical
l a b o r a t o r y conditions showed a s u d d e n fall in
b l o o d sugar level, which h o w e v e r rose again on
withdrawal of the drug, (Table 1).

Conclusions
T h o u g h mitosis in 13 cells is possible, 8 no docu-

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Volume 77, Number3, July 1988

mented report of drug-induced 13 cell

regeneration seems to be available. This is the
first study of its kind with a homeeopathic drug.
Cephalendra indica O. However, the present
observations are in close conformity with the
findings of Chakravarthy et al. with Pterocarpus
marsupium Roxb., 9,1~ which causes selective
regeneration of 13cells of alloxan-damaged pancreas and protects the 13cells against the necrotic
effect. There is a close parallelism between the
slow and steady fall in blood sugar level and
perceptible increase in 13cell counts and graded
inactivation of the hypothalamic neurons (hypotrophy) in the Cephalendra indica 0 treated
group, thus, the in-vitro studies suggest a probable neuroendocrinological mechanism of
Cephalendra indica 0 in stabilizing the blood
sugar level.
The experimental and clinical potential of this
novel antidiabetic drug is specially significant
because it did not show acute or sub-acute toxicity even in fairly large doses, i.e. 0.1 ml to
0.5 ml/100 gbw. Blood pressure screening has
shown hypotensive response at a dose level 0.1
ml/100 gbw. However, we have evaluated the
stability of blood sugar levels for eighteen days
after withdrawal of the drug. Further investigations in this area will be rewarding in order to
confirm neuroendocrinological involvement
through hormonal assay of corresponding hypophysial component.
Acknowledgements
The authors offer their grateful thanks to Dr P. N. Verma,
Director, Homoeopathic Pharmacopoeia Laboratory and Pro-

ject Officer, Drug Standardisation Unit, Ghaziabad, for

extending laboratory facilities and technical support.
References

1 Anderson JW, Herman RH. Treatment of reactive hypoglyc~emia with sulfonylureas Am J Med Sci 1971; 261:
16-23.
2 Clayman CB. Tolbutamide revisited. J A m MedAss 1974;
228: 1523.
3 Widstrom A, Cerasi E. On the action of tolbutamide on
man. I. Role of adrenergic mechanisms in tolbutamide
induced insulin release during normoglyc~emia and
induced hypoglyc~emia. Acta Endocrinol (Copenh). 1973;
72: 506-518.
4 Williams RH, Porte D. The pancreas. In Textbook of
Endocrinology 5th ed, pp. 502-626. (Williams RH ed.)
Philadelphia: WB Saunders Co. 1974.
5 Hartroft WS, Wrenshall GA. Correlation of beta cell granulation with extratable insulin of the pancreas. Studies in
adult human diabetics and nondiabetics. Diabetes 1955; 4:
1-7.
6 Van Wyck JJ, Underwood LE, Hintz RL, Voina SJ,
Weaver RP. Chemical properties and some biological
effects of human somatomedin. In Advances in Human
Growth Hormone Research (Raiti s. ed.) Department of
Health, Education, and Welfare Publication No. (NIH)
74-612. US Government Printing Office, Washington, DC,
1973: 25-45.
7 Agarwal GC, Bapat SK. Indian J Med Res 1977; 66: 323330.
8 Ham AW, System of Endocrine Glands Histology. 7th edn,
p.837. Philadelphia and Toronto: J. B. Lippincon Company 1974.
9 Chakravarthy BK, Gupta S, Gambhir SS, Gode KD, Panacreatic beta cell regeneration--a novel antidiabetic mechnism of Pterocarpus marsupium, roxb. lnd J Pharrn 1980;
12: 123-127.
10 Chakravarthy BK, Gupta S, Gambhir, SS, Gode KD, The
prophylactic action of (-)epicatechin against alloxan
induced diabetes in rats. Life Sci 1981; 29: 2043-2047.