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Pancreatic beta-cell regeneration
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
~ u .~
~
eq~l I
~u - ~ +l
~.= .=
,--~ r,~ en +l
o It -
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
P
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-
151
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.
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
x
o +1 cqC
~rq..
oos
~ +1
+I
ao +I
e4
-
+l
X
"2.
o's
t-- r!,
- e~
~+l
~ll
~m+l
C~+l
~+1
r q "2.
od +1
X
~
~'~ c-I ~
+1
o~
c4 +1
~ ~
. II
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
~ u .~
~
eq~l I
~u - ~ +l
~.= .=
,--~ r,~ en +l
o It -
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
P
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-
151
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.