Effect of Lindane (δ- isomer) on Serum cholesterol and Liver enzyme markers in mice (Mus musculus)

 

Pimpi Sahu¹, Kumar Kritartha Kaushik^1,2, Sanjeeb Kumar Nath³

¹Department of Zoology, Gauhati University, Gopinath Bordoloi Nagar, Guwahati, Assam, India - 781014.

²Assistant Professor, Department of Zoology, Sipajhar College, Darrang, Assam, India - 784145.

³Associate Professor, Department of Botany, Dhing College, Nagaon, Assam, India - 782123.

 

ABSTRACT:

In the present study, we investigate the serum cholesterol level and the liver enzymes (alkaline phosphatase, alanine transaminase, and aspartate aminotransferase) in mice. Lindane (δ- isomer), 100mg/kg body weight, injected subcutaneously for 30 days. Our present study found that after 30 days of lindane administration, after sacrificing the mice, a significant increase in serum cholesterol levels observed. Alkaline phosphatase levels significantly decreased, whereas the alanine transaminase and aspartate aminotransferase levels significantly increased.

 

 

INTRODUCTION:

Agriculture is an integral part of India. Pesticides used in the agricultural sector to kill pests on an excellent harvest attaining popularity¹. Use of pesticides can lessen the disease caused by vectors in plants². The toxicity levels in animals increase due to extensive use of pesticides and it is reported that most of the insects become resistant³.

 

Lindane, an organochlorine pesticide used in the agricultural sector⁴. Lindane is lipophilic, and it gets assembled in animals comprising human beings through the food chain⁵. The previous experiment on rodents shows that lindane is hepatotoxic⁶, neurotoxic⁷, and also affects the reproductive system⁸. According to the World Health Organization, lindane is reasonably toxic. Humans are exposed to lindane by eating foods that contain pesticides, whereas another point of supply is drinking water. Lindane acquaintance diarrhea, convulsions along with some other physiological conditions and finally may lead to death^9,10.

 

Inflammation of the digestive tract may occur if poisoned by lindane. The toxicity of lindane starts by exciting the nervous system. Lindane hinders the GABA channels^11-13. Lindane hinders the contraction of the uterus by restricting the myometrial gap junction¹⁴. Lindane alters the homeostasis of glutathione. Oxidative stress increases due to lindane exposure and leads to physiological malfunctioning^7,15,16. Serum cholesterol level helps to detect different underlying physiological conditions¹⁷. There is a considerable probability that high serum cholesterol level leads to cardiac diseases. Therefore it is necessary to lower the increased level of serum cholesterol. Previous experiments in rats have observed that Achyranthes aspera extract given in different doses has antihyperlipidemic properties. Plant extracts are a good alternative as their side effects are less than synthetic drugs¹⁸. Likewise, S. senegambica has hypocholesterolemic activity¹⁹. An experiment in rats showed that fish oil has a hypocholesterolemic functionary²⁰. The main sterol produced by animals is cholesterol present in the membrane of cells²¹, Lipovedic drug found to be anti-hypercholesterolemic that is genotoxicity free²². Liver disease is a worldwide issue. To determine abnormality in liver functions ALT, AST and ALP tests are done. Generally, ALT and AST are most frequently performed to check the level of liver enzymes. AST is found in the cytosol and as well as in mitochondria. On the other hand, ALT is only present in cytosol^23-26.

 

MATERIALS AND METHOD:

Animals: Adult albino mice weighing 28-35gm provided by Zoology Department, Gauhati University, Assam, India.  Mice kept in 12 hr light and 12 hr dark period, relative humidity, 75%-87% and temperature, 27-30·C. Animals acclimatized to normal environmental conditions in the laboratory for two weeks before use. Standard diet (pellet diet) and water ad libitum supplied regularly.

 

Chemicals:

Lindane (δ-HCH) bought from Zenith India Guwahati, Assam, India. Alcohol and chloroform supplied by the Department of Zoology, Gauhati University, Assam, India.

 

Doses:

100mg/kg body weight dose prepared in 1ml of ethanol and 9ml of distilled water. 0.1ml of the dose was injected subcutaneously for 30 days.

 

Grouping of animals: Eighteen adult mice divided into three groups- control, vehicle control, and lindane injected mice. The weight of the mice was 28-35grams.

 

Blood collection:

Serum separated from blood by centrifugation method, blood is taken out for estimation by cardiac puncture.

Estimation of cholesterol, ALP, AST, and ALT: Using coral, tulip kits all the parameters estimated.

 

RESULTS:

The cholesterol level significantly increases in lindane induced mice compared to control. However, the rise of cholesterol level in-vehicle control is not significant. The ALP level significantly decreased compared to control, and AST and ALT levels significantly increased compared to control. The values of the cholesterol, ALP, AST, and ALT shown in the table below.

 

Table: Cholesterol, ALP, AST, and ALT values of lindane induced mice.

The values expressed in Mean ± SEM. Values are significant at P<0.05 level compared to control values determined by one way ANOVA analysis.

 

DISCUSSION:

Lindane hinders GABA receptors which surge cardiotoxicity²⁷. Liver microsomes, mitochondria, fluidity of the plasma membrane²⁸ and sarcoplasmic reticulum in rat decreased in previous experiments²⁹. In testes, liver and brain oxidative breakdown of lipids take place due to lindane^6,7,30. Lindane incites cell damages by creating ROS³¹. In a study in female Wistar rats it is found that the cholesterol level significantly increases, ALP level significantly decreases and AST and ALT level significantly increases³¹. Lindane causes hepatotoxicity previously documented³². In earlier experiments serum cholesterol level found to be increased in mice as well as in rats due to lindane impaired food^33-35.

 

Our present study shows that lindane alters the cholesterol level as well as alters the liver enzyme markers. Our study accords with previously documented results.

 

Pesticides are useful to kill the pests in the agricultural field. It indeed increases the yield of agricultural products, but in the long term, it harms the animal world, including human beings, by entering into the food chain. In humans the accumulation of lindane affects the vital organs. So, it is high time to switch to and search for a plant-based product that will resist pests in agriculture and the long-run without harming the health.

 

ACKNOWLEDGEMENTS:

We would take the opportunity to thank the Head of the Department of Zoology, Gauhati University for providing all the facilities including animal house to take proper care of mice during entire course of experiment to perform the experimental work. Kumar Kritartha Kaushik is thankful to the Department of Science and Technology, India. Kumar Kritartha Kaushik is a recipient of DST-INSPIRE Fellowship.

 

CONFLICT OF INTEREST:

Authors declared that there is no conflict of interest.

 

PLAGIARISM REPORT:

Manuscript has no (0%) plagiarism while checked on Grammarly.

 

REFERENCES:

1.     Abhilash PC. Singh N. Pesticide use and application: an Indian scenario. Journal of Hazardous Materials. 2009; 165 (1): doi.org/10.1016/j.jhazmat.2008.10.061

2.     Templeton DJ. Jamora N. Economic Assessment of a Change in Pesticide Regulatory Policy in the Philippines. World Development.2010; 38(10): doi.org/10.1016/j.worlddev.2010.06.009

3.     Goudey-Perrière F. Lemonnier F. Bergougnoux V. Perrière C. Low doses of the pesticide lindane induce protein release by the fat body of female cockroach Blaberus craniifer (Dictyoptera). Comparative Biochemistry and Physiology Part C: Toxicology and Pharmacology. 2007; 146 (4): doi.org/10.1016/j.cbpc.2007.06.001

4.     Solomon LM. Fahrner L. West DP. Gamma Benzene Hexachloride Toxicity: A Review. Archives of Dermatology. 1977; 113 (3): 10.1001/archderm.1977.01640030099018

5.     Report W. International programme on chemical safety. In: Health and Safety Guide-54 Lindane (WHO, Geneva). 1991; 14

6.     Videla LA. Barros SBM. Junqueira VBC. Lindane-induced liver oxidative stress. Free Radical Biology and Medicine. 1990; 9 (2): doi.org/10.1016/0891-5849(90)90120-8

7.     Bano M. Bhatt D. Neuroprotective role of a novel combination of certain antioxidants on lindane (g-HCH) induced toxicity in cerebrum of mice. J Res J Agric Biol Sci. 2007; 3 (6)

8.     Chitra K. Sujatha R. Latchoumycandane C. Mathur P. Effect of lindane on antioxidant enzymes in epididymis and epididymal sperm of adult rats. J Asian J Androl. 2001; 3(3)

9.     Hrnčić D. Rašić-Marković A. Djuric D. Šušić V. Stanojlović O. The Role of nitric oxide in convulsions induced by lindane in rats. Food and Chemical Toxicology. 2011; 49(4): doi.org/10.1016/j.fct.2010.12.019

10.  Zucchini-Pascal N. de Sousa G. Rahmani R. Lindane and cell death: At the crossroads between apoptosis, necrosis and autophagy. Toxicology. 2009; 256(1): doi.org/10.1016/j.tox.2008.11.004

11.  Hfaiedh N. Murat J-C. Elfeki A.Protective effects of garlic (Allium sativum) extract upon lindane-induced oxidative stress and related damages in testes and brain of male rats. Pesticide Biochemistry and Physiology. 2011; 100 (2): doi.org/10.1016/j.pestbp.2011.03.009

12.  Bist R. Bhatt DK. The evaluation of effect of alpha-lipoic acid and vitamin E on the lipid peroxidation, gamma-amino butyric acid and serotonin level in the brain of mice (Mus musculus) acutely intoxicated with lindane. Journal of the Neurological Sciences. 2009; 276(1): doi.org/10.1016/j.jns.2008.09.008

13.  Ogata N. Vogel SM. Narahashi T. Lindane but not deltamethrin blocks a component of GABA-activated chloride channels. The FASEB Journal. 1988; 2(13): doi.org/10.1096/fasebj.2.13.2458984

14.  Maranghi F. Rescia M. Macrì C. Di Consiglio E. De Angelis G. Testai E. Farini D. et al. Lindane may modulate the female reproductive development through the interaction with ER-β: an in vivo–in vitro approach. Chemico-Biological Interactions. 2007; 169(1): doi.org/10.1016/j.cbi.2007.04.008

15.  Saravanan M. Prabhu Kumar K. Ramesh M. Protective effect of gallic acid against lindane induced toxicity in experimental rats. Food and Chemical Toxicology. 2011; 49(4): doi.org/10.1016/j.fct.2011.01.005

16.  Vijaya Padma V. Sowmya P. Arun Felix T. Baskaran R. Poornima P. Association of Height with Total Cholesterol Level in Young Adults in Assessment of Coronary Heart Disease Risk. Research Journal of Pharmacy Technology. 2018; 11(3): 10.5958/0974-360X.2018.00155.5

17.  Latif NA. Firdous J. Mamat SNZ. Mona R. Haque A. Muhamad N. Antihyperlipidemic activity of Achyranthes aspera Linn leaves on cholesterol induced hyperlipidemia in rats. Research Journal of Pharmacy Technology. 2017; 10(1):10.5958/0974-360X.2017.00043.9

18.  Sarvesh C. Fernandes J. Janadri S. Yogesh H. Swamy S. Weight Reducing and Hypocholesterolaemic Effect of Aqueous Leaf Extract of Sansevieria senegambica Baker on Sub-Chronic Salt-Loaded Rats: Implication for the Reduction of Cardiovascular Risk. Research Journal of Pharmacy Technology. 2011; 4(5).

19.   Ikewuchi CC. Ikewuchi JC. Ayalogu EO. Onyeike EN. Weight Reducing and Hypocholesterolaemic Effect of Aqueous Leaf Extract of Sansevieria senegambica Baker on Sub-Chronic Salt-Loaded Rats: Implication for the Reduction of Cardiovascular Risk. Research Journal of Pharmacy Technology. 2011; 4(5)

20.  Arunagiri T. Pandian A. Ramesh S. Hypocholesterolemic Effect of Dietary Fish Oil in Albino Wistar Rats. Research Journal of Pharmacy Technology. 2019; 12(9): 10.5958/0974-360X.2019.00718.2

21.  Mallikarjuna Rao N. Bagyalakshmi J. Ravi T. Development and Validation of an Analytical Method for the Estimation of Standard Cholesterol and Egg Cholesterol by High Performance Thin Layer Chromatography (HPTLC). Research Journal of Pharmacy and Technology. 2011; 4(1)

22.  Sumanth M. Swetha S. Narasimharaju K. Natesh T. Genotoxicity Testing of Lipovedic-A Polyherbal Anti-hypercholesterolemic Drug. Research Journal of Pharmacy Technology. 2011; 4(8)

23.  Edokwe Chinelo OO. Joshua Parker Elijah. Biochemical Effects of Persea americana (Avocado) and Dacryodes edulis (African Pear) Extracts on Liver Function Test of Albino Wistar Rats. Research Journal of Science and Technology. 2010; 2(2)

24.  Sawarkar D. Vijaya C. Turaskar A. Shende V. Chatap V. Sawant V. Borkar SN. Hepatoprotective Activity of Ethanolic and Ethyl Acetate Extract of Ziziphus mauriatiana on Liver Damaged Caused by Paracetamol in Rats. Research Journal of Pharmacognosy Phytochemistry. 2009; 1(3)

25.  Rajesh M. Latha M. Hepatoprotective Activity of Glycyrrhiza glabra Linn. on Experimental Liver Damage in Albino Rats. Research Journal of Pharmacognosy Phytochemistry. 2010; 2(4)

26.  Ghawate V. Shrivastava P. Bhambar R. Hepatoprotective activity of Bridelia retusa bark extracts against carbon tetrachloride-induced liver damage rats. Research Journal of Pharmacognosy Phytochemistry. 2017; 9(2): 10.5958/0975-4385.2017.00022.X

27.  Sauviat MP PN. Cardiotoxicity of Lindane- a gamma isomer of hexachlorocyclohexane. J SocBiol .2002;196

28.  Bagchi M. Stohs SJ. In vitro induction of reactive oxygen species by 2,3,7,8-tetrachlorodibenzo-p-dioxin, endrin, and lindane in rat peritoneal macrophages, and hepatic mitochondria and microsomes. Free Radical Biology and Medicine.1993;14(1): doi.org/10.1016/0891-5849(93)90504-N

29.  Antunes-Madeira MC. Almeida LM. Madeira VMC. Depth-dependent effects of DDT and lindane on the fluidity of native membranes and extracted lipids. Implications for mechanisms of toxicity. Bulletin of Environmental Contamination and Toxicology. 1993; 51(6): 10.1007/BF00198271

30.  Samanta L. Sahoo A. Chainy G. Age-related changes in rat testicular oxidative stress parameters by hexachlorocyclohexane. Archives of Toxicology. 1999; 73 (2)

31.  Sharma P. Shankar S. Agarwal A. Singh R. Variation in serum lipids and liver function markers in lindane exposed female wistar rats: attenuating effect of curcumin, vitamin C and vitamin E. Asian J Exp Biol Sci. 2010; 1(2)

32.  Ravinder P. Srinivasan K. Radhakrishnamurty R. Dietary hexachlorocyclohexane induced changes in blood and liver lipids in albino mice. Indian Journal of Experimental Biology New Delhi. 1990; 28(2)

33.  Boll M. Weber LW. Stampfl A. The effect of γ-hexachlorocyclohexane (lindane) on the activities of liver lipogenic enzymes and on serum lipids in rats. Zeitschrift für Naturforschung C. 1995; 50(1-2): doi.org/10.1515/znc-1995-1-220

34.  Boll M. Weber LW. Stampfl A. The response of rat serum lipids to diets of varying composition or contaminated with organochlorine pesticides. Zeitschrift für Naturforschung C. 1996; 51(1-2): doi.org/10.1515/znc-1996-1-216

35.  Ricardo KFS. Oliveira TTd. Nagem TJ. Pinto AdS. Oliveira MGA. Soares JF. Effect of flavonoids morin; quercetin and nicotinic acid on lipid metabolism of rats experimentally fed with triton. Brazilian Archives of Biology Technology. 2001; 44(3): doi.org/10.1590/S1516-89132001000300007

 

 

Accepted on 04.02.2022           © RJPT All right reserved

Research J. Pharm. and Tech 2022; 15(12):5721-5723.