| Abstract|| |
Lactovedic is a lactogenic polyherbal formulation containing Jivanti, Shatavari, Vidarikanda, Yashtimadhu and Shatapushpa, and is processed with swarasas of Brahmi, Mandukaparni, Matsyakshi, Shatavari and Kokilaksha. The aim of this study was to evaluate the galactagogue activity of lactovedic. Rats (175-200 g) suckling eight to nine pups were divided into four groups (n=6). Control group rats were treated with vehicle (2 ml of 1% carboxymethyl cellulose sodium in normal saline) orally, Group II and Group III rats were orally administered 270 and 540 mg/kg body weight, respectively, of lactovedic suspended in vehicle, and Group IV animals were treated orally with 2.7 mg/kg body weight of domperidone suspended in vehicle from 3rd day of parturition to 15th day of parturition. Milk yield at 18 hours, the weight of pups at 18 and 23 hours and the daily weight of the mother rat were estimated. On 16 th day, blood samples were collected and mother rats were sacrificed. Glycogen and total protein content in mammary gland and serum prolactin and cortisol were estimated. Results were statistically analysed using analysis of variance (ANOVA), followed by Tukey-Kramer post hoc test. Histopathology of mammary gland was performed. Lactovedic increases the milk yield, pups' body weight, weight of the mother rat, glycogen and protein content of mammary gland tissue, and serum prolactin and cortisol, compared to the control animals. Transverse section of mammary gland of lactovedic treated rats showed proliferation of acini and marked increase in milk secretion in the ducts. It can be concluded that lactovedic possesses significant galactagogue activity.
Keywords: Cortisol, galactagogue, histopathology of mammary gland, lactovedic, milk yield, prolactin, pups weight
|How to cite this article:|
Sumanth M, Narasimharaju K. Evaluation of galactagogue activity of lactovedic: A polyherbal formulation. Int J Green Pharm 2011;5:61-4
| Introduction|| |
Mother's milk is important for survival, proper development and growth of the neonate. Milk is the only source of water, organic nutrients and minerals, to which the neonates have access. Colostrum (the first milk taken from the mammary gland after parturition) and mature milk contain non-nutrient substances (such as antibodies and bioactive factors) that are important for growth, development and survival of the neonate. Low supply of milk is one of the most common reasons given for discontinuing breast feeding. Galactagogues are medications or substances to assist initiation, maintenance and augmentation of maternal milk production.  Ayurvedic medicines based on ancient scripts like Sushrutha Samhitha, Charaka Samhitha, etc., are herbal formulations. Some of the herbal drugs having galactagogue activity are Leptadenia reticulate,  Asparagus racemosus,, Ipomoea digitata, Glycerrhiza glabra,, Centella asciatica, Bacopa monnieri, Anethum sowa, etc. Vedic Bio Labs, Bangalore, has developed lactovedic 500 mg capsule, a natural milk enhancer which contains extracts of Jivanti, Shatavari, Vidarikanda, Yashtimadhu and Shatapushpa. The chemical constituents and medicinal uses of these plants are given in [Table 1]. The present study was taken up to generate preclinical data to support the clinical use and hence standardisation of lactovedic as a galactagogue. The galactagogue activity of lactovedic was evaluated by estimation of milk yield, pups' body weight, weight of the mother rat, glycogen and protein content in mammary gland tissue, and estimation of serum prolactin and cortisol.
|Table 1: Chemical constituents and medicinal uses of plants in polyherbal formulation lactovedic|
Click here to view
| Materials and Methods|| |
Wistar female rats weighing 175-200 g, procured from Drug Testing Laboratory, Bangalore, were maintained under standard laboratory conditions (25±2°C and RH 60±5%) with free access to food and water ad libitum. Our institute's animal house registration no. is 152/99/CPCSEA/1999. Rats were divided into four groups of six animals each. Control group rats were treated with vehicle (2 ml of 1% carboxymethyl cellulose sodium in normal saline) orally, Group II and Group III rats were orally administered 270 and 540 mg/kg body weight, respectively, of lactovedic suspended in vehicle, and Group IV animals were treated orally with 2.7 mg/kg body weight of domperidone suspended in vehicle, from 3rd day of parturition to 15th day of parturition. The dose of lactovedic was calculated from human dose (as given in the ancient scripts). Institutional Animal Ethics Committee's permission was obtained before starting the experiments.
Milk Yield and Pups' Body Weight
Every day during the study period, 13 hours after the treatment of mother rat, the pups were weighed (w1) and subsequently isolated from their mother for 4 hours. At the end of 4 hours, the pups were weighed (w2), returned to their mother and allowed to feed for 1 hour and were weighed (w3) again. Subsequently, pups were isolated from their mother for 4 hours and then weighed (w4), following which they were reunited with their mother for 1 hour of feeding and, finally, they were weighed (w5). They were subsequently left with their mother during the night. Milk yield at 18 and 23 hours after the gavage was estimated as (w3 - w2) and (w5 - w4), respectively. Daily milk yield at 18 hours was corrected for weight loss due to metabolic processes in the pup (respiration, urination and defaecation) during suckling. The value used was (w2 - w1)/4. This value was multiplied by the number of suckling hours per day and added to the daily suckling gain. Daily weight gain of pups was calculated from the pup weight at w2. 
Weight of the Mother Rat
Daily weight of mother rat was taken and the difference of weight between 3 rd and 15 th day after parturition was determined.
Estimation of the Glycogen and Protein Content Mammary Gland Tissue
On 16 th day, blood samples were collected and mother rats were sacrificed. Mammary gland was isolated and cleared from the other connective tissue and weighed accurately using electronic balance. About 100 mg of tissue was homogenised in 1.5 ml of distilled water using tissue homogeniser (Remi Motors RQ127A, Mumbai, India), mixed with 1.5 ml of 30% KOH saturated with Na 2 SO 4 and heated for 30 minutes in boiling water bath, cooled and centrifuged (Remi C24) after adding 2 ml of 95% ethanol. The precipitated glycogen was collected from the alkaline digestate, dissolved in distilled water and estimated by phenol-sulphuric acid method. 
Total protein content  was estimated using total protein kit (Span Diagnostic Ltd., Bangalore, India).
Serum Prolactin and Cortisol Estimation
On 16th day after parturition, the blood was collected from retro-orbital sinus, serum was separated, the prolactin and cortisol were estimated by using electrochemiluminiscence (Rat Prolactin ELISA kit, Catalog Number RPRL.96 of MD biosciences, USA) and enzyme immunoassay (Cortisol EIA kit, Cayman chemicals, Mumbai, India), respectively. 
Histopathology of Mammary Gland
On 16th day, mother rats were sacrificed, mammary gland was isolated and cleared from the other connective tissue and processed for histopathological studies. The histological sections were projected on a white glazed paper through a projection microscope; the parenchyma area was counted against the mammary stroma. The microslides of mammary gland were examined using the projection microscope at 2500× magnification for scoring the secretary rating.
The data were presented as mean±SEM. Results were statistically analysed using analysis of variance (ANOVA), followed by Tukey-Kramer post hoc test, using Graph Pad InStat version 3.00, Graph Pad Software, CA, USA. The level of significance was set at P<0.05
| Results and Discussion|| |
As shown in [Table 2], lactovedic significantly increases milk yield, pups' body weight, mother rat's weight, serum prolactin and cortisol compared to the control animals. During the lactating period, the mean milk yield at 18 and 23 hours was increased [Figure 1]. Lactovedic increases protein content, glycogen, parenchyma percentage and secretary rating of mammary gland tissue [Table 3].
|Figure 1: Effect of lactovedic on mean milk yield at 18 and 23 hours; values are expressed as mean±SEM (n=6); ANOVA; Tukey-Kramer post hoc test; ***P<0.001|
Click here to view
As shown in [Figure 2], transverse section (TS) of mammary gland of lactovedic treated rats showed proliferation of acini and marked increase in milk secretion in the ducts (back to back arrangement of cells, indicating increased secretory rate), as compared to normal control and standard domperidone treated animals.
|Figure 2: Effect of lactovedic on mammary gland; (a) TS of mammary gland of control rat, showing closely packed lobuloalveolar tissue; (b) TS of mammary gland of lactovedic treated rat, showing proliferation of parenchyma. acini and enhanced milk secretion in alveoli|
Click here to view
One of the many reasons for discontinuing breast feeding is low supply of milk in lactating mothers. Lactovedic is an Ayurvedic polyherbal galactagogue, which contains Jivanti, Shatavari, Vidarikanda, Yashtimadhu and Shatapushpa, and is processed with swarasas of Brahmi, Mandukaparni, Matsyakshi, Shatavari and Kokilaksha. Some of the medicinal plants identified as lactogenic stimulate the synthesis of lactogenic hormones (prolactin, growth hormone, cortisol) and/or β-endorphin and β-casein accumulation in the mammary gland. After parturition, prolactin stimulates the synthesis of milk proteins in the epithelial cells and proliferation of secretory cells. , Our studies indicate that lactovedic increases serum prolactin, protein content and glycogen of mammary gland. Histopathological studies reveal that lactovedic increases proliferation of acini, lobuloalveolar size, parenchyma percentage and milk secretion in the ducts. The measurement of milk yield in lactating rats is done indirectly by means of pup weight. The increase in milk yield in our studies is due to the increased serum prolactin, which stimulated the biosynthesis of milk. ,
| Conclusion|| |
To conclude, it can be said that lactovedic, a polyherbal formulation, possesses significant galactagogue activity.
| References|| |
|1.||Sjolin S, Hofvander Y, Hillervik C. Factors related to early termination of breast feeding: A retrospective study in Sweden. Acta Paediatr Scand 1977;66:505-11. |
|2.||Available from: http://www.himalayahealthcare.com/herbfinder/h-leptad.htm#c [last accessed on 2008 Nov 18]. |
|3.||Goyal RK, Singh J, Lal H. Asparagus racemosus - An Update. Indian J Med Sci 2003;9:407-14. |
|4.||Available from: http://www.holisticonline.com/Herbal-Med/_Herbs/h203.htm [last accessed on 2008 Nov 18]. |
|5.||Moharana D. Shatavari. Jastimadhu and Aswagandha: The ayurvedic therapy. Orissa Review: Bhubaneshwar. 2008. p. 72-7. |
|6.||Kokate CK, Purohit AP, Gokhale SB. Drugs containing glycosides. Pharmacognosy. 39 th ed. Pune: Nirali Prakashan; 2007. p. 212-20. |
|7.||Available from: http://www.himalayahealthcare.com/herbfinder/h-shatapush.htm [last accessed on 2008 Nov 18]. |
|8.||Ouedrago ZL, Heide DV, Beek EM, Swarts HJ, Mattheij JA, Sawadogo L. Effect of aqueous extract of Acacia nilotica ssp adansonii on milk production and prolactin release in the rat. J Endocrinol 2004;182:257-66. |
|9.||Joanne TE, Jack CB, Mina JB. Interrelationship of glycogen metabolism and lactose synthesis in mammary epithelial cells of mice. Biochem J 1980;192:695-702. |
|10.||Lowry OH, Rosebrough NJ, Farr AL, Randall RR. Determination of protein content using phenol reagent. J Biol Chem 1951;193:265-75. |
|11.||Ahmadzadeh A, Barnes MA, Gwazdauskas FC, Akers RM. Dopamine antagonist alters serum cortisol and prolactin secretion in lactating holstein cows. J Dairy Sci 2006;89:2051-5. |
|12.||Sawadogo L, Houdebine LM, Thibault JF, Rouau X, Olivier-Bousquet M. Effect of pectic substances on PRL and growth hormone secretion in the ewe and on the induction of casein synthesis in the rat. Reprod Nutr Dev 1988;28:293-301. |
|13.||Brisken C, Kaur S, Chavarria TE, Binart N, Sutherland RL, Weinberg RA, et al. PRL controls mammary gland development via direct and indirect mechanisms. Dev Biol 1999;210:96-106. |
|14.||Lockwood DH, Turkington W, Topper YJ. Hormone- dependent development of milk protein synthesis in mammary gland in vitro. Biochim Biophys Acta 1966;130:493-501. |
|15.||Turkington RW, Brew K, Vanaman TC, Hill L. The hormonal control of lactose synthetase in the developing mouse mammary gland. J Biol Chem 1968;243:3382. |
Department of Pharmacology, Visveswarapura Institute of Pharmaceutical Sciences, 22nd Main, 24th Cross, B.S.K II stage, Bangalore - 560 070, Karnataka
[Figure 1], [Figure 2]
[Table 1], [Table 2], [Table 3]