Ancient Science of LAfe, Vol, IIT, No. 2, October RESPONSE OF COMMIPHORA MUKUL (GUGGULU) ON MELATONIN INDUCED HYPOTHYR’ iM A. K. SINGH, 8. N. TRIPATHI and G, C. PRASAD Department of Kayachikitsa and Shalya Shalyakya, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221005, India, Received March 28, 1983, Accepted: Fuly 2, 1903, ABSTRACT: ‘The results of this study indicate that melatonin induces hypothyrol- dism in mice within eight days. Simultaneous administration of melatonin and potroloum ether extract of Commiphora mukul brings about @ significant improve- ment in thyroid structure and function. Further, melatonin induced hypothy- roidism is although a little reversible yet it is normalized quickly after C. tiukul ‘treatment. ‘These date suggest that C, mukul @irectly stimulates thyroid fanction probably through some enzymatle mechantams, Introduction Our earlier studies on Comiphora mukul have revealed a direct thyroid stimulating property in thiouracit induced hypothyroi- sism (under publication). Besides, reports ae also available where its therapeutic claims are suggestive of its action through tiycoid gland and neomercazole induced Iypercholesterolemia has been reported to regress by the use of this drug!. Further, yuthogenesis of bypercholesterolemia in hypothyroidism is also. an established fic®3. In view, to confirm the above observations an experimental study was undertaken to find out the effect of Commiphora mukul on experimental hypo= thyroidism induced by melatonin. Material and methods In this study 180 young male mice (6-8 week) were maintained on the labora- tory environmental conditions including light, temperature, diet and water available ad libitum. All the animals were divided in tne following groups for different treatments: (i) First group of 108 animals were admini- sered intraperitoneally synthetic melatonin | in the dose of 2Srhg/100 gm body weight daily for eight days and the suppression of 85 thyroid function was assessed on every alternate days by studying the functional and structural status of thyroid gland. At the end of 8th day melatonin treatment was discontinued and the remaining 72 animals were further divided into following two subgroups-a36 animals of one group were administered intraperitoneally — petroleum, ether extract of C. mek! in the dose of 20 mg/l00_gm of body weight, (b) while the remaining 36 animals were administered the vehicle solution and served as control group. @) The second group of 36 mice received a combined treatment of melatonin 5 mg/100 gm body weight) as well as P. E. extract of C. mukul (20 mg/100 gm body weight) by the same route. G) Remaining last group of animals served as normal and were kept on saline water throughout the experimental period. Nine animals from each group were sacrificed by decapitation in between 10-11 A.M. and were subjected to the study. of thyroid functions in three batches: (a) Thyroid 1" uptake: Four animals ‘of each group were intraperitoncally injec- ted 5 uci radioactive 1''=24 hours beforesacrifice. The thyroid glands of these were dissected out and 24 hours 1 8! uptake) thyroid lobe was determined. (b) Thyroid hormones assays: Five animals of each group were anaesthetized with ether and they were bled from orbital sinus. The blood was pooled and serum was subjected for the estimation of T;- resin uptake* and scrum PBI. Free thyro- xine index was also determined by the miethod of Clark’, (¢) Histological study: ‘Thyroid gland of five mice were dissected out, fixed in houins fixative, sections were cut at $ micron thickness, and stained with hematoxylin and cosin. In addition the cell height was also measured with the help of an occulo- micrometer. Results (a) Preventive response of C. mukul: The results of this study revealed nearly no change in the weight and I uptake of thyroid gland in the normal while a significans (pZ_0.001) decline was observed in the melatonin treated group. However, in the melatonin and C. muku treated group of animals there was a significant (p/ 0.001) increase in the weight as well as 13! uptake of thyroid gland, »Ty-resin uptake was also decreased which ‘was maximum on the 4th day. The animals receiving combined treatment of melatonin and C. mukul showed an increase in T;—resin uptake which was low on the 2nd day but was higher as compared to control in the later period. Further, a continuous and signi- ficant fall in the serum PBI and free thyroxine index was observed in the melatonin treated group which was rever- sed with this combined administration of C. mukut and melatonin, (Table ~ 1). Histological observation of thyroid land of melatonin treated group revealed reduction in the follicular cell height which appeared thin and squamous at the end of Sth day and the colloid material was thick and inactive. On the other hand combined treatment of C. mukul and melatonin to animals brought about a significant increase in the follicular cell height which was maximum on the 4th day. Hypertrophy and_ hyperplasia of the follicular cells were also associated with the appearance of vacuolization in the colloid material (Plate and Table ~ 1), (©) Curative response of C. mukul: Results of this study showed an increase the weight and 1" uptake of thyroid gland after the withdrawal of melatonin treatment, However, a significant (p/_0.001) increase in the weight and 1! uptake of thyroid gland was observed when the animals were administered —C, _mukut ‘Ty-resin uptake after melatonin treatment withdrawal was observed to increase from 2rd day to 8th day but a highly significant {®Z.0.001) increase was observed alice C.-mukul treatment. Further, a steady crease: in serum PBI and free thyroxine index was also observed when the melatonin treatment was stopped but increase was highly significant (p/ 0.001) when C. muda! was administered (Table-2). Histological examinations of thyroid after discontinuing melatonin treatment showed a continuous increase in follicular cell height and it coincided with control value on the 8th day, The colloid material was homogenously spread in the lumen. However, in C. mukul treated group of animals there was a sharp incresse in the cell height which was maximum on the 4th day. Follicullar hypertrophy and hyper plasia were observed in the early period but reduced by the end of 8th day and only few columnar cells were present. (Plate and Table-2) Discussion Direct response of melatonin on thyroid function seems to be controversial. However, it has been reported by many workers to have antithyrcdial effects and it brings about a decrease in thyroid metabolism and hormone production’, Melatonin treatmeat to rodents has been formed to cause 2 reduction in thyroid weight, secretion and 1 uptake *°. This study has also confirmed the earlier observations'67, Such func. tional changes are also suggestive of | hypothyroidism. The structural changes | 86,“Bs ‘TABLE-1 Preventive effect of C. mukul on melatonin induced hypothyroidism (Mean £S.D.) DAYS AFTER TREATMENT ‘Thyroid Functions 1 I Vv I WI ‘vit c Ty roe Ty nm | G¢ Tv hh Cc Th T 1331 uptake (%) 953 874 11.95 10.67 9.48 902 127 19.92 £106 41.31 4.1.63 £1.74 £193 0.91 £1.52 £1.87 T; resin uptake % 58.53 47.14 52.59 53.39 56.63 $7.83 4632 62.07 £132 £321 £215 £2.70 43.62 £5.01 4161 41.93 Serum PBI (ug!) 10.2 86 = 13.03 10.6 90 63 1839 9S $8 20.41 £03 $$ 4195 414 ALI 416 4272 426 40.21 41.94 Free Thyroxine Index 10.6 7.14 12.37 10.0 845 4.58 21.78 99.2 4.64 $103 £0.56 11.2 £209 411 4097 409 412 4071 Cell height (Microns) 8.25 6.15 1525 9.21 5.09 1868 9.82 3.12 1425 10.25 2.91 13.25 4102 41.52 £141 1098 41.24 £1.38 41.09 40.93 £1.09 41.32 1.0.79 41.24 © =Controt; ‘T) ~ Melatonin treated: y= Melatonin and C. mulul treated * P/_0.08 as compared to control ** P/_0,02 ax compared to control *** P/0.01 as compared to control **** PZ 0.001 as compared to control‘TABI Curative effect of C, mukul on LE - IT ‘melatonin induced hypothyroidism (Mean +. S. D.) ces DAYS APTER TREATMENT oid — i W Vi Val en € F c T € T c a Wtuptake = «8.38 12.SL 9.29 15.97 10.15 10.95 1635 ) 4043 £085 4115 4167 40.92 1.05 2 Ty-resin 49.81 S491 50.35 60.95 52.76, $4.98 63.23 uptake % = £1.98 £2.66 L173 £335 41.92 $2.73 12.72 £1.95 Serum PBI 6.51 9.55.67) 12.23 7.86 S61 «9.231829 (ug %) £0.45 420.66 0.32 098 30.75 £125 4041 LL Frecthyro- 5.89 8.59 GIL 13.57 7.05 992 1615 xine index. 0.75 40.97 86 1.28 4.0.85 10.96 1.89 Cellheight 4.12 7.28 «= 4.99 12.62 5.63 695 9.32 (micron) £1.26 41.62 41.31 $1.34 11.82 41.78 © = Treated control; T= C. mukul treated "a *p/_0.08 as compared to control 'PZ_0.01 as compared to control **p/_0.02 as compared to control —_****p/ 0.001 as compared to control clucidate that reduction in the weight and follicular cell height of thyroid might have been imparted due to inhibitary response of melatonin via pituitary 12” However, there is difference in opinion regarding the level of action of melatonin on pituitary, 134 Our earlier study has indicated that melatonin directly inhibits thyroid function through specific melatonin binding protein in thyroid glands, Further, simultaneous administration of C. mukui alongwith melatonin neutra- lizes the inhibitory action of melatonin both structurally and functionally. This indicates that probably C. mukul might be activating the pituitary to stimulate thyroid functions provided melatonin depresses TSH secretion causing hypothyroidism. In case, the depressant effect of melatonin is confirmed in spite of raised TSH under this condition, it is most likely that C. mukul might be hampering the inhibitary action of melatonin causing a direct stimutatory action on thyroid gland. However, the latter sugge- stion seems to be stronger as melatonin directly supresses thyroid functions (Singh etal, 1981). Although the natural recovery in thy- roid structure and function has been found only by the withdrawal of melatonin but CC. mukul has significantly increased the thyroid functions and morphology. Thus, our results suggest that C. mukul stimulates thyroid directly probably through some or the other enzymatic mechanisms and may prove to be a good treatment in specific type of hypothyroidism in goitre. Acknowledgement : Authors are thankful to the Director General, ICMR for peoviding the necessary funds. We are obliged to Prof. K.N.Udupa for his kind suggestions. 88LEGENDS Homogenous collold alter B. After melatonin us epithe- lium enclosing thick and inactive colloid material. C. After combined tre. fatmont of melatonin as well as P. E, extract of G. Muka, Sowing’ the fol cular hypertrop! Gnd vacudlated cor (H&E xX 400)6 Tripathi, S. N., Gupta, M., Sen, S. P. and Udupa, KN, ind. Jour. Exp. Biol., 1975: 135 15. Kutly, K. M., Byrant, D. G., Farid, N. RJ: J. Clin Endocr, Metals, 1978, 46; Mathe, D., Chevallier, F. : Biochem Biophys. Acta, + 1976, 441: 155, Clark, F. : Lancet, 1963, 2 : 167. Acland, J. D. + Biochem. J, 1957, 66:17 Clark, F. and Horn, D. Bo: J. Clin Endocrinol Metals. 1965, 25 ; 39. Cardinati. D. : Current topies in experi- mental Endocrinology. New York. Acad. press 1974, 2: 107. Baschieri, L., Deluca, F., Cremarossa, L., Demartina O., Olivierio. A. 2 Negri Mz: Experientia, 1963, 19 = 15. 90 . Relkin, R. De Prospo, N. D., Demartino, 1.5. ¢ al. : Life Seience, 1968, 7 : 183. |. Csaba, G., Reti L, Fisher J. : Acta Med. Aead. Sci. Hung., 1970, 27: 183. » Reiter, R. J.: Frachinis, F. + Newroendo- crinology, 1969, 5: 219. Neuroendocrinology 1972, 10: 46. Relkin, R. : Neuroendoerinology, 1978, 25:310. Brammer, G. L., Morley, J. E., Geller, E.G., Yuwiler, A. and Hershman, 1M LAM. J. Physiol, 1979, 236: E Singh, A. K. and Prasad. G, C. Jour. Med. Res., 1981, 74 : 420. Vriend. J. and Reiter, R. J. : Metab. Res., 1977,9 : 331 Yriend, J., Reiter, R. J. and Anderson, G. Ry Gen. Comp. Endocr. 1979, 38 189, Horm.