Tuberculostatic Activity of Henna (Lawsonia

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Tubercle (1990) 71, 293-29.50 Longman Group UK Ltd 1990

Tuberculostatic activity of henna (Lawsoniainemis Linn.)V. K. SHARMA*S. M. S. Medical College, Hospital for Chest and Tuberculosis, Jaipur, India

Sum ma ry-The tuberculostatic activity of the herb henna Maw sonia ir terrnis Linn.) was testedin vitro an d in viva. On Lowenstein Jensen med ium, the growth of tubercle bacilli from sputumand of Mycobacferium tuberculosis H3 , Rv wa s inhibited by 6 yglml of the herb. I n v i v a studieson guinea pigs and m ice showed that the herb at a dose of 5 mg/kg body weight led tosignificant resolution of experimental tuberculosis following infection with M. tuberculosisH3 ,Rv.

Introduction Materials and methodsThe discovery of the tubercle bacillus by Robe rt Preparat ion o f the herbal drugKoch in 1882 intensified the search for an effec- oh e dtive and specific drug against tuberculosis. Today, rug was prepared from henna (La w s o n i amany bactericidal and bacteriostatic drugs are inermis Linn.), a shrub indigenous to the Arabicused in combination all over the world with dra- peninsula and Iran and widely cultivated in India.matic positive results and tuberculosis has A total of 250 g of powdered leaves was added tobecome a curable disease. Even with the advent 1 1 of water, boiled for 15 min over low heat,cooled and filtered. The filtrate w as reduced toof such powerful SpCCifiC antimicrobials, a longduration Of treatment with either daily Or inter- l/3 o f its original volume by heating on a watermittent therapy is required . In the presently avail- bath. It was stored ready for use in airtight bottlesable regimens of 6-9 months duration, the and its weight/ml was measured against water.patients frequently stop taking drugs as soon asthe symptoms are ameliorated and the treatmentis therefore discontinued. We therefore wish to In v i t r o s t u d yfind a specific drug which can cure tuberculosis in Five sputum samples, positive for tubercle bacilli,a much shorter time. In this study we report the were inoculated on to five drug-free control slo-tubercu lostatic effect of a prepa ration derived pes of Lowenstein Jensen (LJ) and on to anotherfrom henna (La w s o n i a i n erm i s Linn.), i n v i t r o on five LJ slopes containing the preparation at a con-Lowenstein Jensen medium and i n v i v o in guinea centration of 6 ug/ml and incubated for 6 weeks.pigs and mice. In another study, Mycobacter ium tuberculos i sstrain Hs7R v was likewise inoculated on to five*Correspondence to: Professor V. K. Sharma, 4 Ja 6, Jawahar drug-free and five drug-containing LJ slopes andNagar, Jaipur - 302004, India incubated for 8 weeks.

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29 4 SHARMAIn vivo studyA total of 20 guinea pigs with an average weightof 500 g (range 490-606 g) were inoculated intra-muscularly in the groin with 0.2 ml of a suspen-sion containing 30000 M. tuberculosis HX7Rv bymean s of a tuberculin syringe. The numbe r ofbacilli was calculated by McF arlands nephelo-meter stand ards for bacterial counts [l]. The ani-mals were kept in four separate cages, five ani-mals to a cage. Two cages served as controls. Theanimals in the experimental group were given thepreparation of henna in a dose of 5 mg/kg bodyweight intramuscularly daily for 15 days startingfrom the day when the first of the control animalsdied. All these animals were kept under observa-tion for 15 weeks from the date of inoculation.Anim als, in either group, that survived for 15weeks were killed and their viscera were sent forhistopathological examination. In another study,40 mice were inoculated with the same dose ofH37R ~ in the tail. Four animals were kept in eachcage. Mice surviving for 40 days were given thedrug, 5 mg/kg body weight daily by intramuscularinjection, for 1 month from the 40th day of inocu-lation. The m ice were kept for a period of 15weeks.

ResultsIn vitroAfter 6 weeks incubation all control media slopeshad more than 2 0 colonies with the typicalappearance of tubercle bacilli. On microscopicexamination, these colonies were acid-fast. Noneof the slopes containing 6 pg/rnl of henna showedany bacterial growth after 6 weeks and the waterof condensation was also negative for acid-fastbacilli.

In vivoGuinea pigs: Weight loss was observed in all theguinea p igs: the weight dropped to an average of300 g (range: 25@ --350 g) over 2 weeks. All ani-mals became sluggish in their activity from thethird day of inoculation and their appetite wasmarkedly reduced. Patchy loss of hair and erec-tion of hair follicles a ll over the body wa sobserved from the 5th day of inoculation inalmost all animals. All 10 control animals diedwithin 5 weeks of inoculation: i.e. on days 23,24(2), 26 (2), 27, 28, 31 (2) and 33. On macroscopic

Mice: All the mice were sluggish in their activityfrom the second w eek onw ard. Loss of hair andappetite was observed in all animals and in fourcases there was an ulcer at the site of inoculation.A total of 10 animals died d uring the first 35 days;i.e. on days 21 (2), 22 (3), 33 (2), 34 (2) and 35.The remaining 30 animals were treated from the40th day for a period of 30 days. All these animalssurvived for a period of 15 weeks, after whichthey were killed. Nine of these an imals hadmacroscopical and microscopical evidence oftuberculosis (Table 1).

DiscussionIn 1945 Florey [2] reported some experiments onthe activity of certain antibiotics on experim ental

Table 1 The numbers of untreated and treated micewith tuberculous lesions and the numbers of animalswith various organs involved

Organs involvedTotal Mice with LymphGroup mice lesions Lung Kidney nodes Liver

Untreated 10 10 (100%) 9 10 10 9Treated 30 9 (30%) 6 4 9 4

examination, tubercles w ere observed on the sur-face of spleen, liver and lungs and the lymphnodes showed necrotic areas. On microscopicexamination caseating necrosis with epithelioidand giant cells were observed in all tissues.In the experimental group, in which therapywas started on day 23, all 10 animals startedimprov ing by the fourth day of the initiation ofdrug therapy. They regained their appetite and insome cases it was increased so that it was neces-sary to serve more food. They gained weight withmost regaining their normal weight of around500 g within 3 weeks. The weight of three animalsexceeded their pre-experimental weight: one ani-mal weighed 600 g and two animals w eighed morethan 560 g. The rem aining seven returned to theirnormal weight of 500 g by the end of the 3 weeks.Hair loss ceased and all animals became activeagain within 4 weeks. All survived for 15 weeks,after which they were killed and their viscerawere examined histopathologically. There was nomacroscopic or microscopic evidence of tubercu-losis.

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TUBERCULOSTATIC ACTIVITY OF HENNA

tuberculosis. n none of these experimentsweretuberculous lesions of animal arrested, though thegrowth of tubercle bacilli was inhibited in culture.The first drug which arrested tuberculous lesionsin guinea pigs was the compound diamino-diphenyl sulphone (dap sone). Later, after discov-ery of streptomycin by Wa ksm an, quantitativemethods for determining the tuberculostaticactivity of antibiotics were developed [3].Henna has been used to treat skin infectionssuch as Tinea and it is known to have a ntibacterialproperties which have been attributed to naph-thoquinones, including lawsone [4]. Several otheragents with activity against the tubercle bacillushave been isolated from plants. Japanese workersisolated alkaloids from a vine named S t ep h o n i au p h a r a n t h a , and from a wistaria like plant re-ferred to as S. sasaki i , and the alkaloid capha-ranthine has been used in the treatment andprophylaxis of tuberculosis in Japan [5]. Chinesework ers investigated the activity of a series oflocal plants against the tubercle bacillus andWa ng [6] reported prominent tuberculostaticactivity in an extract of coptis root (Copt i s ch i -mens i s ) . This activity lies in the alkaloid berberinsulphate. In the Indian medical traditions ofAyurveda and Unani, many plants are consideredto be useful in the treatmen t of tubercu losis [7,8].In addition, A erous ca lam us , Alp in ia ga langa,Cucurbi ta maxima, Pepos i s , Eucalyp tus c i t ro idorsan d Crew la popul ipho l ia are known to have someactivity against M. tuberculosis [S]. Some otherplants and plant-derived materials w ith suchactivity have been described by Grange and

29 5Davey [9]. These include Actuea sp ica tu , Gal ipeaoff icinal is , Pip er cubeba, several resins and pro-polis (bee glue).In general, though, the use of herbs to treattuberculosis, although attempted in many coun-tries, ha s not met with success. By contrast, thisstudy has revealed that an aqueous extract ofhenna has strong i n v i t r o an d i n v i v o tuberculosta-tic acitivity. F urther evaluation of this herb istherefore indicated.

References1.

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Woffls G. McFarlands nephelometer standards for bacte-rial counts. In Sonnenwirth AC, Jarett L, eds. Grud-w ohls, clinical laboratory methods and diagnosis. 8th ed.,Vol. 2. St. Louis: CV Mosby Co. 1980.Florey HW. Use of micro-organisms for therapeutic pur-poses (Lister memorial lecture). Brit Med J 1945; 2: 635-642.Hinshaw HC, Garland LH. D&eases of the Chest. Phila-delphia: Saunders Co.Wren RC. Potters new cy clopaedia o f botanical drugsand preparations. Revised edition p 143. Saffron Walden:CW Daniel Co. Ltd. 1988.Nishikawa H. Screening tests for antibiotic action of plantextracts. Jap J Exp Med 1949; 20: 337-349.Wang VFL. In vitro antibacterial activity of some com-mon Chinese herbs on Mycobacterium tuberculosis. Chi-nese Med J 1950; 68: 169-172.Chopra RN, Chopra IC, Handa KL, Kapur LD, eds.Choprass Indigenous Drugs of India 2nd ed. Calcutta:UN Dhur & Sons Pvt, Ltd 1958.Indian Council of Medical Research. Medicinal Plants oflndia Vol. 1 and 2. Delhi: ICMR 1976.Grange JM, Davey RW. Detection of antituberculousaetivity in plant extracts. J Appl Bact 1990; 68: 587-591.