ORIGINAL�ARTICLE ABSTRACT Objective: To appraise the preventive effect of turmeric on anti-mosquito coil smoke induced interstitial fibrosis in lungs of Sprague Dawley rats. Study Design: Randomized control trial. st th Place and Duration of Study: This study was completed in 10 months from 1 August 2014 to 28 May 2015; conducted at Islamic International Medical College, Rawalpindi and National Institute of Health (NIH), Islamabad, Pakistan. Material and Methods: Three equal groups of 21 adult albino rats were made. Control Group X was retained in fresh air. Experimental Groups M & T inhaled Mosquito Coil smoke for 12 weeks. Group T also had oral turmeric 300mg/kg body weight. After 12 weeks rats were dissected, and their lung tissues were studied microscopically for the presence of fibrosis. Results: Interstitial Fibrosis was markedly present in the histological sections of lung tissues from experimental group M showing thickened fibrotic alveolar walls and inflammatory cell infiltrate with reduced air spaces in all animals (100%) in group M. Fibrosis was present in 2 out of 7 rats in group T showing a significant protection with only 28.5% of animals with fibrotic lungs. No interstitial fibrosis (0%) was seen in the lung tissues of animals in control group X; all 7 rats showed well-formed alveoli. Conclusions: Interstitial lung fibrosis caused by Anti-Mosquito coil smoke can be protected with the use of turmeric. Key Words: Covid-19, Curcuma, Mosquito Coil, Pulmonary Fibrosis, Turmeric. nutraceutical. Turmeric has significant anti- 3 inflammatory and anti-oxidant properties. It has gained value as a digestive aid, treatment for fever, wounds, infections and blood disorders. Data from animal and pharmacological studies also supports that active ingredients in turmeric play a protective role in various diseases like acute respiratory distress syndrome, chronic obstructive pulmonary disease and allergic asthma and also have proven antiviral properties against para-influenza type 3, respiratory 4,5,6 syncytial virus and herpes simplex virus, its therapeutic action is based on the inflection or prevention of oxidative stress and inflammation confirmed with the studies done at molecular & 7,8 cellular levels. The findings in these researches give substance to the possibility of testing turmeric for the prevention of pulmonary fibrosis caused by allethrins released from burning anti-mosquito coils. Mosquitoes and the transmission of the mosquito borne diseases have been historically controlled by the use of chemical insecticides and mosquito repelling agents like anti-mosquito coil containing Pyrethroids (allethrins), which play a central role in Introduction Modern medicine to a large extent has failed in its ambition to control both acute and chronic diseases. Traditional herbal medicines have been found effective for the treatment and prevention of several 1 diseases. Turmeric is a traditional herbal medicine. It's broad reaching mechanism of action and lack of systemic toxicity may make it best suited as an adjuvant therapy for various viral and other respiratory disorders like tobacco smoke induced epithelial damage and acute lung injury that are 2 resistant to currently available therapies. Several diseases have been found to have inflammatory origin. Turmeric is safe, affordable and efficacious Preventive Effect of Turmeric against Anti-Mosquito Smoke Induced Pulmonary Fibrosis in Laboratory Rat Saira Jawed, Wafa Omer 1 2 Correspondence: Dr. Saira Jawed Assistant Professor Department of Anatomy HBS Medical and Dental College, Islamabad E-mail: sairajawed371@gmail.com 1 2 Department of Anatomy /Pathology HBS Medical and Dental College, Islamabad Funding Source: NIL; Conflict of Interest: NIL Received: February 11, 2020; Revised: October 01, 2020 Accepted: October 10, 2020 Preventive Effect of TurmericJIIMC 2020 Vol. 15, No.4 222 9 mosquito control programs. More toxic products having higher concentration of allethrins are predominating the market due to the rapid spread of 10 resistance worldwide. The use of allethrins in higher concentrations and a complicated mixture of metallic vapors to tackle the resistant mosquitoes, 11 exposes the consumer to a severe health risk. It has already been addressed in the literature that exposure to mosquito coil smoke can have toxic effects on the lungs and can induce histopathological changes like severe emphysema, hyperplasia, chronic obstructive pulmonary disease, edema, hemorrhage & fibrosis, necrosis, hyperemia, connective tissue infiltration by inflammatory cells and obstruction by hyaline material within the lung 12,13 tissue. The objective of the current research was to demonstrate the efficacy of turmeric in preventing pulmonary fibrosis as there is insufficient literature on the preventive characteristics of turmeric against anti-mosquito allethrins present in coil smoke. Material and Methods This randomized control trial conducted at Islamic International Medical College, Rawalpindi and National Institute of Health (NIH), Islamabad and was completed in 10 months. A total of 21 Sprague Dawley male rats weighing 250-300g were purchased from the animal house of National Institute of Health (NIH) Islamabad and were randomly divided into three groups; control group X and experimental groups M & T. The study was performed after an ethical approval from the Institutional Review Committee of Riphah International University. Female rats and the rats having a weight of less than 250 g or having a physical disability were excluded from the study. Groups M & T rats were placed in smoke exposure cabins at a temperature of 27±3ºC with a 12hr light/dark cycle along with standard laboratory diet and water. Experimental groups were exposed to mosquito coil smoke for 7 hours per day for 12 weeks; group T received 300mg/kg body weight oral turmeric as well. All rats were sacrificed at the end of 12 weeks and left lungs of all were dissected out and preserved in 10% formalin. After tissue processing slides were prepared and stained with Masson's Trichrome to be studied microscopically first at low power, x10, and then at high power, x40 objective. The histological slides were interpreted by the pathologist. Slides were studied for presence of interstitial fibrosis. Masson's Trichrome stained collagen fibers causing thickening of alveolar septa & alveolar spaces filled with proteinaceous fluid and inflammatory cell infiltrate causing reduction in alveolar spaces was taken as a criterion for interstitial fibrosis. Results The lung tissue of all rats (100%) in group M (Fig 2A) had a Masson's trichrome stained dense fibrous connective tissue with collagen fibrils & thickening of alveolar walls with cellular infiltration, proliferation & airspaces filled with proteinaceous fluid. The interstitial infiltrate consisted of lymphocytes, plasma cells, mast cells and eosinophils resulting in interstitial fibrosis. Interstitial pulmonary fibrosis was present in 2 out of 7 rats in group T; thus rats in group T showed a remarkable protection from pulmonary fibrosis with only 28.5% of animals developing fibrosis (Fig 2B). The result is shown in table (Fig 1). The Masson's Trichrome stained histological slides from the rats in control group X had normal honeycomb like alveolar structure with well-formed alveolar walls (Fig 2C). Table I: Table represen�ng percentage of inters��al lung fibrosis in control and Experimental Groups JIIMC 2020 Vol. 15, No.4 223 Preventive Effect of Turmeric Discussion Pulmonary fibrosis is a pattern of reaction to parenchymal damage in which lung has unusually high collagen fiber content per unit volume of 14 tissue. In the present study histological sections of the lungs of rats are stained with Masson's Trichrome in order to observe the fibrotic thickening of alveolar septa, reduced air spaces and presence of proteinaceous fluid. 100% of rats in Group M showed thickened alveolar walls & proteinaceous fluid accumulated in interstitial space along with interstitial fibrosis. In Group T, only 28.5% of rats showed the said histopathological parameters. Inflammation is one of the etiological factor for the 14 development of fibrosis. As inflammatory reaction is observed in this case, it can be correlated to the development of fibrosis. Fibrosis in the pulmonary interstitium is usually a result of alveolar damage due 14 to physical, chemical or microbial agents. Allethrin in the coil smoke can be linked to the interstitial fibrosis observed in this study. The process of interstitial fibrosis is characterized by thickened alveolar walls that are fibrotic and contain an inflammatory cell infiltrate with reduced air spaces. Macrophages are thought to be responsible for expressing fibroblast growth factor, resulting in the proliferation of fibroblasts and subsequent development of fibrosis. The excessive deposition of collagen fibers reflects reduced lung capacity leading to respiratory distress which is the main effect of air pollutant exposure. Earlier study mentions that smoke induces lung destruction with inflammatory reaction, more fibroblasts are brought to the irritated area leading to more collagen fiber 15,16 deposition. In another study, fibrotic change was observed in lung parenchyma of rats after 6 weeks of exposure to mosquito coil smoke. Similar histological appearance was seen in the lungs exposed to 17 asbestos and cigarette smoke. Excessive deposition of collagen fiber has also been observed in the lungs 18 of rats in similar inhalational studies. Turmeric is one of the herbal compounds that has been 19,20 investigated in fibrosis research. In the present research only 28.5 % of histological sections of rat lungs in the Group T showed fibrous reaction. The result of this study is consistent with the previous studies. Turmeric administration can inhibit 21 abnormal lung collagen formation. Turmeric supplementation suppresses alveolar macrophages, expression of fibroblast growth factor by the macrophages and other inflammatory cells thereby ameliorating the inflammatory responses in the 22 lungs which prevents fibrosis in the lung tissue. The histopathological findings of the current research can further be utilized to test turmeric for the prophylaxis and treatment of Covid-19 induced pulmonary fibrosis; which is considered to be the 23 most devastating outcome of this viral infection. Conclusion Interstitial lung fibrosis caused by anti-mosquito coil smoke can be protected with the use of turmeric. Fig 2: (A) Histological Sec�on of a Rat Lung from Group M Showing Thickened Fibro�c Alveolar Septa (Arrow), Proteinaceous Fluid (Star) With Reduced Air Space (Double Headed Arrow) (B) Histological Sec�on of a Rat Lung from Group T Showing Less Thickened Alveolar Septa (Arrow) With A Few Cellular Infiltrates (Star), Well-Formed Alveoli and Preserved Air Space (Double Headed Arrow) (C) Histological Sec�on of A Rat Lung from Group X Showing Normal Alveolar Septa (Arrow) and Alveoli Showing Honeycomb Like Pa�ern (Double Headed Arrow) JIIMC 2020 Vol. 15, No.4 224 Preventive Effect of Turmeric REFERENCES 1. Kant S, Lohiya A, Ahamed F, Abdulkader RS, Singh AK, Silan V. Comparative morbidity profile of patients attending an Ayurveda clinic and a modern medicine clinic of a primary health center in rural Haryana, India. J Fam Med Prim care. 2018;7(2):374. 2. 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