|Year : 2017 | Volume
| Issue : 6 | Page : 1500-1504
|Tanacetum parthenium relieves chronic constriction injury-induced neuropathic pain in male rats
Hossein Ali Safakhah1, Masoumeh Tatar2, Ali Ghanbari3
1 Department of Physiology, Faculty of Medicine; Research Center of Physiology, Semnan University of Medical Sciences, Semnan, Iran
2 Student Research Committee, Faculty of Medicine, Semnan University of Medical Sciences, Semnan, Iran
3 Research Center of Physiology, Semnan University of Medical Sciences, Semnan, Iran
Click here for correspondence address and email
|Date of Web Publication||11-Jan-2018|
| Abstract|| |
Background: Neuropathic pain is a chronic developing pain due to injury or disease-involving somatosensory systems such as vascular disorders, autoimmune diseases, inflammation, and traumatic injuries. Herbal medicines were utilized for the treatment of various diseases too late. The aim of this study was to evaluate the effect of Tanacetum parthenium (TP) extract on chronic constriction injury (CCI)-induced neuropathic pain. Materials and Methods: In the present study, 48 adult male Wistar rats weighing 200–250 g were used. After anesthetizing the animals, CCI was performed on the left sciatic nerve for inducing neuropathic pain. TP extracts were intragastrically administered daily for 2 weeks by a gavage tube. Mechanical allodynia and thermal hyperalgesia were assessed using Von Frey hairs and plantar test device, respectively. The data were analyzed using the one-way analysis of variance and Tukey's post hoc test. Results: CCI led to mechanical allodynia and thermal hyperalgesia. TP extract with higher doses used in this study significantly increased (P < 0.001) paw withdrawal threshold in response to mechanical stimulation. Further, it increased (P < 0.01) paw withdrawal latency in response to thermal stimulation. Conclusion: Chronic orally used TP extract alleviates CCI-induced neuropathic pain in male rats.
Keywords: Chronic constriction injury, neuropathic pain, rat, Tanacetum
|How to cite this article:|
Safakhah HA, Tatar M, Ghanbari A. Tanacetum parthenium relieves chronic constriction injury-induced neuropathic pain in male rats. Ann Trop Med Public Health 2017;10:1500-4
|How to cite this URL:|
Safakhah HA, Tatar M, Ghanbari A. Tanacetum parthenium relieves chronic constriction injury-induced neuropathic pain in male rats. Ann Trop Med Public Health [serial online] 2017 [cited 2019 Sep 23];10:1500-4. Available from: http://www.atmph.org/text.asp?2017/10/6/1500/222657
| Introduction|| |
Central and peripheral nerve injury causes neuropathic pain. Neuronal injury is followed by various disorders such as diabetic neuropathy, amputation, viral infection, spinal cord injury, and stroke. Different animal models have been presented for studying neuropathic pain in which chronic constriction injury (CCI) is one of the most utilized so far. Available data show that a combination of central and peripheral mechanisms including anatomical, neurochemical, and excitotoxic changes are involved in neuropathic pain due to sciatic nerve injury.,, Sciatic nerve injury not only causes peripheral dysfunction, but also leads to cortical and subcortical changes through the brain stem. Persistent interruption of peripheral nerve or nervous inflammation usually leads to abnormal processing of painful and nonpainful stimuli that result in phantom limb pain, allodynia, and hyperalgesia. Neuropathic pain affects different aspects of patient's life. Treating the pain is very complicated and it includes interventional and noninterventional approaches. Regardless of a considerable measure of drugs, neuropathic pain is a difficult situation to treat and no certain cure exists so far.
Plants have been used for the prevention and treatment of diseases from ancient time and still in use worldwide. Plants and natural sources are bases of new medicines and help us to provide commercial drugs. About 25% of prescribed drugs in the world are obtained from plants. Feverfew herb (Tanacetum parthenium L [TP]) belongs to the Asteraceae family, 30–100 cm in height, and has yellow-green flowers. Orally used or injection of extracts of feverfew is useful in several inflammatory diseases. It is also reported that feverfew is useful to treat constipation, fever, headache, menstrual disorders, stomach ache, tinnitus, toothache, and vertigo., Jain et al. have reported that orally used feverfew leaf extracts significantly increase tail withdrawal response to thermal stimulus in rats. Furthermore, it has been reported that orally used feverfew extracts from the flowers and the leaves reduce fever and inflammation. Past studies have revealed the effectiveness of the chronic use of feverfew extracts in different disorders such as migraine and inflammatory conditions including arthritis and asthma., A review of past studies suggests that feverfew has beneficial effects on pain, but there is no research on CCI neuropathic pain. The aim of this study was to evaluate the effect of feverfew leaf extract on CCI neuropathic pain in male rats.
| Materials and Methods|| |
In this study, adult male Wistar rats weighing 200–250 g were used. The rats were provided from animal house of Semnan University of Medical Sciences and housed in a room with controlled humidity, temperature (20°C ± 2°C), and 12 h light-dark cycle. All rats had free access to food and water. All experimental procedures were performed according to local ethical committees on animal research of the Semnan University of Medical Sciences, under permit number 93/584231.
Induction of neuropathy
Neuropathy was induced through the method introduced by Bennett and Xie. After anesthetizing the animals with intraperitoneal injection of ketamine (80 mg/kg) and xylazine (10 mg/kg) mixture, the fur on the left thigh was shaved, and using a surgical blade, a 2-cm incision was made. After exposing the sciatic nerve, the nerve was released from the surrounding connective tissues. Four loose ligations were made around the common sciatic nerve at 1-mm intervals using catgut chromic sutures 4.0. The ligations should not disrupt nerve blood flow. In the end, incised muscle and skin were sutured with silk 4.0 separately. In the sham group, muscle and skin were sutured without manipulation of the sciatic nerve.
Hydroalcoholic extract of TP was provided from Barij Essence Company (Kashan-Iran).
Experimental protocol and groups
In this study, 48 male rats were randomly divided into six groups (intact, sham, neuropathy, neuropathy with TP extract 10 mg/kg, neuropathy with TP extract 20 mg/kg, and neuropathy with TP extract 40 mg/kg). TP extract was orally given in a volume of 2 ml by a gavage tube.
The experimental design is shown in [Table 1].
Assessment of pain response to mechanical stimulus
Animals' pain behavior was evaluated using the following tests.
Paw withdrawal response was detected using the method described by Ren  with von Frey's filaments (Stoelting, Wood Dale, IL, USA). Calibrated filaments were applied with ascending style on the dorsal surface of the left hind paws. Each filament exerts a defined force (in grams) as a stimulus. The test began with the minimum force, and in case of no response, the greater force was examined. If animal pulls its foot off, between three and five stimuli are delivered, and that force was considered as a response. The stimulus of 60 g was considered as a cutoff force.
After habituating the animal in the plantar test device, infrared radiation with intensity of 60 was applied to the plantar surface of the injured paw in three consecutive times with 5-min interval. The latency of reaction to thermal stimulus was recorded and the average of three times was considered as a response. Cutoff point of the test was 60 s.
GraphPad Software, Inc.7825 Fay Avenue, and Suite 230 La Jolla, CA 92037 USA was used for analyzing the data. The data were analyzed using the one-way analysis of variance and Tukey's post hoc test. All data were expressed as mean ± standard error of the mean of response. P < 0.05 was considered statistically significant.
| Results|| |
In the present study, sensitivity of the injured paw to both of tactile and thermal stimuli was evaluated. Generally, to evaluate the development of neuropathic pain, tactile allodynia and thermal hyperalgesia were tested by measuring paw withdrawal response.
Mechanical allodynia was assessed using von Frey filaments applied on the paws' dorsal surface between the second and third toes. [Figure 1] shows response threshold of the lesioned paw to mechanical stimulus. Paw withdrawal response in the CCI group was reduced (P < 0.05) significantly against that in the intact group. TP extract (40 mg/kg) significantly increased (P < 0.001) paw withdrawal response compared to CCI group. Response to mechanical stimulus in the sham group was not different from those in the intact group.
|Figure 1: Effects of different doses of Tanacetum parthenium extract on mechanical allodynia. Response threshold to tactile stimulus was significantly reduced in chronic constriction injury group with respect to intact group. Tanacetum parthenium extract (40 mg/kg) significantly increased paw withdrawal response compared to chronic constriction injury group. Data were expressed as mean ± standard error of the mean of measured variable (n = 8)|
Click here to view
We evaluated paw withdrawal latency time to thermal stimuli using plantar test device.
The time between initiation of radiation and hind paw lifting or jumping was used as thermal hyperalgesia threshold. [Figure 2] shows response threshold of the lesioned paw to a thermal stimulus. Paw withdrawal latency time in the CCI group was significantly (P < 0.05) lower than those in the intact group. TP extract significantly increased (P < 0.01) paw withdrawal latency compared to CCI group. Paw withdrawal latency time does not make significant difference between sham and intact groups.
|Figure 2: Effects of different doses of Tanacetum parthenium extract on thermal hyperalgesia. Paw withdrawal latency to thermal stimulus was significantly reduced in chronic constriction injury group against that in the intact group. Tanacetum parthenium extract (40 mg/kg) significantly increased paw withdrawal response compared to chronic constriction injury group. Data were expressed as mean ± standard error of the mean of measured variable (n = 8)|
Click here to view
| Discussion|| |
TP is among the various herbs used in the remedy of different disorders such as migraine, inflammation, and rheumatism. In the present study, the effect of hydroalcoholic extract of TP on neuropathic pain due to CCI of the sciatic nerve in the male rat was evaluated. Results showed that CCI reduces the threshold of paw withdrawal to mechanical and thermal stimuli against that in the intact group but not in sham group. A possible explanation for this might be related to gavage procedure used in our experiments. Several reports have shown that orogastric gavage procedure is a stressful procedure , and it activates hypothalamic-pituitary-adrenal axis. It was also discovered that gavage-induced stress leads to apoptosis of liver cells  and even execution of gavage procedure in a sham group led to similar physiologic results. Therefore, in our study, a reasonable explanation for the nonsignificant difference between sham and treated groups may be related to gavage procedure that caused some pain in sham animals. Our results showed that 14-day gavage of TP extract (40 mg/kg) significantly decreases neuropathic pain. It has been reported that acute pretreatment with TP extract significantly alleviates acetic acid injection-induced visceral pain in mice that is inhibited using naloxone. Furthermore, reports have shown that TP reduces carrageenan-induced pain  and the observed effect with respect to the parthenolide content of the TP. There are several studies that parthenolide through IκB kinase complex (activator of nuclear factor kappa B [NF-κB]) has a prominent role in signaling of pro-inflammatory cytokines , so that it inhibits tumor necrosis factor (TNF)-alpha production. TNF-alpha leads to neuronal hyperexcitability and increases pain sensation through phosphorylation of N-methyl-D-aspartate (NMDA) receptors in pain circuits. Smolinski and Pestka reported that parthenolide significantly decreases interleukin-6 production in the lipopolysaccharide-treated mice. Previously, we showed that pro-inflammatory cytokine TNF-alpha increases significantly following neuropathic pain in CCI-induced male rats. Therefore, it is possible that reduced pain observed in the present study is a result of TNF-alpha inhibition by parthenolide content of TP.
It has been reported that 7-day intraperitoneal administration of parthenolide inhibited d-fenfluramine-induced neuronal serotonin release in the rat gastric fundus  and even orally administered TP powder was more effective in stimulated serotonin inhibition. Considering the effect of TP inhibition of serotonin release and serotonin involvement in the pain control (gate theory), in this study, involvement of serotonin in the effect of TP in reducing neuropathic pain is impossible.
Lipeng Dong in 2013 reported that parthenolide significantly reduces middle cerebral artery occlusion induced-brain ischemic injury and edema through suppressing the expression of Caspase-1, NF-kB, and p38MAPK. On the other hand, it has been reported that CCI leads to neuronal edema and injury., Therefore, decreased mechanical allodynia in our experiments is probably due to suppression effect of TP on CCI-induced edema on the sciatic nerve. A study showed that systemic injection of parthenolide prominently alleviates face and hind paw mechanical allodynia in the rats whose dura mater had stimulated with TRPA1 agonist. Experiments on the mice revealed that TRPA1 receptors have a role in CCI-induced neuropathic pain. Depending on these reports, another possible mechanism for reduced mechanical allodynia in our study may be through inhibition of TRPA1 receptors. TP flower extract which has higher parthenolide content than the TP leaf extract enabled in suppressing mechanical hyperalgesia in the diabetic male rats. Consistent with this result, our study showed that higher doses of TP (40 mg/kg) significantly increased paw withdrawal threshold response to mechanical stimulus. A study of Galeotti et al. revealed that single oral dose of TP extract attenuates mechanical hyperalgesia in the diabetic male rats and this effect was maintained for 30 min. This short-term result suggests possible direct and acute analgesic effects of TP extract. In our study, TP extract was used 2 weeks and the day after (day 15), we examined neuropathic pain behavior. We believe that the observed hypoalgesic effect of extract in our study was not an acute effect, but it is probably mediated through activation of long-term mechanisms such as modifying gene expression.
A study by Jain et al. in mice revealed that TP reduces lactic acid-induced pain and inflammation and also significantly increases tail withdrawal latency in the tail flick test. Further reports show that parthenolide prominently reduces thermal hyperalgesia in rats., Consistent with these researches, in our study, hydroalcoholic extract of TP suppressed CCI-induced thermal hyperalgesia. Parthenolide is the main sesquiterpene lactone content of TP., It has been revealed that anti-inflammatory effect of parthenolide depends on NF-kB inhibition  which plays an important role in the prevention of pro-inflammatory cytokine release. Previously, we reported that CCI led to significant increase of thermal hyperalgesia along with TNF-alpha. Accordingly, it is possible in the present study that reduced thermal hyperalgesia is due to inhibition of TNF-alpha production by the parthenolide content in the TP.
| Conclusion|| |
It can be concluded that chronic orally used TP attenuates CCI-induced mechanical allodynia and thermal hyperalgesia in male rats.
We would like to express our deep appreciation to Hossein Hosseini (Engineer in Barij Medicinal Plants Research Center) for gifting us hydroalcoholic extract of TP.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
Financial support and sponsorship
This study was supported by a grant from Deputy of the Research and Technology of Semnan University of Medical Sciences, and the authors hereby appreciate the Deputy.
Conflicts of interest
There are no conflicts of interest.
| References|| |
Zimmermann M. Pathobiology of neuropathic pain. Eur J Pharmacol 2001;429:23-37.
Ji RR, Strichartz G. Cell signaling and the genesis of neuropathic pain. Sci STKE 2004;2004:reE14.
Bennett GJ, Xie YK. A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain 1988;33:87-107.
Finnerup NB, Jensen TS. Spinal cord injury pain – Mechanisms and treatment. Eur J Neurol 2004;11:73-82.
Waxman SG, Hains BC. Fire and phantoms after spinal cord injury: Na+ channels and central pain. Trends Neurosci 2006;29:207-15.
Hains BC, Willis WD, Hulsebosch CE. Serotonin receptors 5-HT1A and 5-HT3 reduce hyperexcitability of dorsal horn neurons after chronic spinal cord hemisection injury in rat. Exp Brain Res 2003;149:174-86.
Brüggemann J, Galhardo V, Apkarian AV. Immediate reorganization of the rat somatosensory thalamus after partial ligation of sciatic nerve. J Pain 2001;2:220-8.
Demarin V, Basić-Kes V, Zavoreo I, Bosnar-Puretić M, Rotim K, Lupret V, et al.
Recommendations for neuropathic pain treatment. Acta Clin Croat 2008;47:181-91.
Wachtel-Galor S, Benzie Iris FF, editors. Herbal Medicine: Biomolecular and Clinical Aspects. 2nd
ed. Taylor & Francis; 2011.
Jain NK, Kulkarni SK. Antinociceptive and anti-inflammatory effects of Tanacetum parthenium
L. Extract in mice and rats. J Ethnopharmacol 1999;68:251-9.
Berry MI. Feverfew faces the future. Pharm J 1984;232:611-4.
Murphy JJ, Heptinstall S, Mitchell JR. Randomised double-blind placebo-controlled trial of feverfew in migraine prevention. Lancet 1988;2:189-92.
Setty AR, Sigal LH. Herbal medications commonly used in the practice of rheumatology: Mechanisms of action, efficacy, and side effects. Semin Arthritis Rheum 2005;34:773-84.
Pittler MH, Ernst E. Feverfew for preventing migraine. Cochrane Database Syst Rev 2004;4:CD002286.
Ren K. An improved method for assessing mechanical allodynia in the rat. Physiol Behav 1999;67:711-6.
Tal M, Bennett GJ. Extra-territorial pain in rats with a peripheral mononeuropathy: Mechano-hyperalgesia and mechano-allodynia in the territory of an uninjured nerve. Pain 1994;57:375-82.
Pareek A, Suthar M, Rathore GS, Bansal V. Feverfew (Tanacetum parthenium
L.): A systematic review. Pharmacogn Rev 2011;5:103-10.
Balcombe JP, Barnard ND, Sandusky C. Laboratory routines cause animal stress. Contemp Top Lab Anim Sci 2004;43:42-51.
Walker MK, Boberg JR, Walsh MT, Wolf V, Trujillo A, Duke MS, et al.
less stressful alternative to oral gavage for pharmacological and toxicological studies in mice. Toxicol Appl Pharmacol 2012;260:65-9.
Roberts RA, Soames AR, James NH, Gill JH, Wheeldon EB. Dosing-induced stress causes hepatocyte apoptosis in rats primed by the rodent nongenotoxic hepatocarcinogen cyproterone acetate. Toxicol Appl Pharmacol 1995;135:192-9.
Asgari A, Parvin N. The analgesic effect of ethanolic extract of Tanacetum parthenium
in acetic acid model. Zahedan J Res Med Sci 2013;15:22-5.
Yin MJ, Yamamoto Y, Gaynor RB. The anti-inflammatory agents aspirin and salicylate inhibit the activity of I (kappa) B kinase-beta. Nature 1998;396:77-80.
Mathema VB, Koh YS, Thakuri BC, Sillanpää M. Parthenolide, a sesquiterpene lactone, expresses multiple anti-cancer and anti-inflammatory activities. Inflammation 2012;35:560-5.
Wang XM, Lehky TJ, Brell JM, Dorsey SG. Discovering cytokines as targets for chemotherapy-induced painful peripheral neuropathy. Cytokine 2012;59:3-9.
Smolinski AT, Pestka JJ. Comparative effects of the herbal constituent parthenolide (Feverfew) on lipopolysaccharide-induced inflammatory gene expression in murine spleen and liver. J Inflamm (Lond) 2005;2:6.
Safakhah HA, Moradi Kor N, Bazargani A, Bandegi AR, Gholami Pourbadie H, Khoshkholgh-Sima B, et al.
Forced exercise attenuates neuropathic pain in chronic constriction injury of male rat: An investigation of oxidative stress and inflammation. J Pain Res 2017;10:1457-66.
Mittra S, Datta A, Singh SK, Singh A 5-hydroxytryptamine-inhibiting property of Feverfew: Role of parthenolide content. Acta Pharmacol Sin 2000;21:1106-14.
Dong L, Qiao H, Zhang X, Zhang X, Wang C, Wang L, et al.
Parthenolide is neuroprotective in rat experimental stroke model: Downregulating NF-κB, phospho-p38MAPK, and caspase-1 and ameliorating BBB permeability. Mediators Inflamm 2013;2013:370804.
Wu FX, Pan RR, Yu WF, Liu R. The anti-nociception effect of dezocine in a rat neuropathic pain model. Transl Perioper Pain Med 2014;1:5-8.
Materazzi S, Benemei S, Fusi C, Gualdani R, De Siena G, Vastani N, et al.
Parthenolide inhibits nociception and neurogenic vasodilatation in the trigeminovascular system by targeting the TRPA1 channel. Pain 2013;154:2750-8.
Pinheiro Fde V, Villarinho JG, Silva CR, Oliveira SM, Pinheiro Kde V, Petri D, et al.
The involvement of the TRPA1 receptor in a mouse model of sympathetically maintained neuropathic pain. Eur J Pharmacol 2015;747:105-13.
Galeotti N, Maidecchi A, Mattoli L, Burico M, Ghelardini C. St. John's Wort seed and feverfew flower extracts relieve painful diabetic neuropathy in a rat model of diabetes. Fitoterapia 2014;92:23-33.
Popiolek-Barczyk K, Kolosowska N, Piotrowska A, Makuch W, Rojewska E, Jurga AM, et al.
Parthenolide relieves pain and promotes M2 microglia/Macrophage polarization in rat model of neuropathy. Neural Plast 2015;2015:676473.
Popiolek-Barczyk K, Makuch W, Rojewska E, Pilat D, Mika J. Inhibition of intracellular signaling pathways NF-κB and MEK1/2 attenuates neuropathic pain development and enhances morphine analgesia. Pharmacol Rep 2014;66:845-51.
Rothwarf DM, Zandi E, Natoli G, Karin M. IKK-gamma is an essential regulatory subunit of the IkappaB kinase complex. Nature 1998;395:297-300.
Knight DW. Feverfew: Chemistry and biological activity. Nat Prod Rep 1995;12:271-6.
Research Center of Physiology, Semnan University of Medical Sciences, Semnan
Source of Support: None, Conflict of Interest: None
[Figure 1], [Figure 2]
| Article Access Statistics|
| Viewed||1113 |
| Printed||52 |
| Emailed||0 |
| PDF Downloaded||18 |
| Comments ||[Add] |