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Effect of Persicaria Odorata Leaves Extract on Symptomatic Alterations

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Effect of Persicaria Odorata Leaves Extract on Symptomatic Alterations Original Article Effect of Persicaria Odorata Leaves Extract on Symptomatic Alterations in Diabetic Neuropathic Rats Induced by Streptozotocin and Chronic Constriction Injury Nataya Sritawan1,3, Sittichai Iamsaard1,3, Kowit Chaiciwamongkol1,3, Jintanaporn Wattanathorn2,3, Nongnut Uabundit1,3* 1Department of Anatomy, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand 2Department of Physiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand 3Integrative Complementary Alternative Medicine (ICAM) Research and Development Group, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand *Corresponding author, e-mail: [email protected] Abstract 1) normal control, 2) DM + Vehicle, 3) DM + Diabetic neuropathy is a serious CCI + Vehicle, 4) DM + Sham + Vehicle, complication of diabetes. It caused by 5) DM + CCI + Ascorbic acid, 6) DM + CCI increase of reactive oxygen species + Gabapentin, 7), 8), and 9) DM + CCI + PO resulting in damaging of nerve fiber 3, 30, or 120 mg/kg BW groups, respectively. functions. This condition affects the life The neurobehavioral examinations were quality of diabetic patients. P. odorata (PO) measured on day 1, 7, 14 and 21 whereas is a Thai-local vegetable and proven to have nerve conduction velocity (NCV) was high antioxidant activity. Therefore, this measured on day 21 only. The results showed study aimed to investigate the effects of PO that PO leaves extract could recover sensory leaves extract on neuropathic symptoms in and motor functions and increased NCV rat model induced by streptozotocin (STZ) value. In conclusion, P. odorata leaves extract and chronic constriction injury (CCI). In is able to attenuate the neuropathic symp- methodology, the experiment male rats toms of sciatic nerve in diabetic rats. were induced to be DM by STZ before CCI Keywords: Diabetic neuropathy, of sciatic nerve. Then the animals were Persicaria odorata, Neuropathic symptoms, divided into 9 groups (n=6). There were Chronic constriction injury, Streptozotocin 76 Vol.11 No.4 1. Introduction oxygen species, especially natural plants Diabetic neuropathy is a common that have antioxidant capacity, has gained complication of diabetes mellitus, affecting extensive attention.7-12 as many as 50% of diabetic patients.1, 2 It is Persicaria odorata (or Polyganum characterized by a progressive loss of pe- odoratum) is a local plant of southeastern ripheral nerve fibers involved in sensory, Asia. P. odorata is classified in family of motor, autonomic or combined neuropa- Polygonaceae and genus of Persicaria. This thies.3 Neuropathic symptoms were also plant can grow up15 to 30 cm. Its leaf is dark described as an abnormality of sensations green and the stem jointed off each leaf. such as dysesthesia (unpleasant abnormal The flavor of P. odorata is described as be- sensation), hyperalgesia (an increased ing pungent and spicy and is commonly response to painful stimuli), and allodynia used in cuisines.13 The previous studies (pain in response to a stimulus that does identified compounds in P. odorata includ- not normally provoke pain).4,5 The ing quercitol, flavonoids, azetidine 2-car- pathophysiology of the neuropathic boxylic acid, mucous polysaccharides and condition remains unclear, although it has steroidal compounds. 14-16 Moreover, P. odo- been associated with the degeneration of rata leaves have been shown to possess myelinated and unmyelinated sensory strong antioxidant activity and there are fibers, and reducing the peripheral nerve many beneficial effects such as antibacte- conduction.2 rial, antifungal and hepatoprotective Previous studies have described that effect.13,15,17,18 However, there is no report hyperglycemia leads to excess formation of about the effect of P. odorata on diabetic reactive oxygen species, induces oxidative neuropathy. Therefore, the purpose of this stress in neurons and activates multiple study was to assess the effect of P. odorata biochemical pathways which these factors leaves extract on diabetic neuropathic play an important role in the development symptoms in rat model induced by strepto- of diabetic neuropathy.6 Therefore, search- zotocin (STZ) and chronic constriction in- ing for effective drugs for treatment dia- jury (CCI) model. betic neuropathy by eliminating reactive 77 Vol.11 No.4 2. Materials and methods induce diabetes mellitus in experimental 2.1 Plant material and extraction animals. 2,19-21 Fresh leaves of P. odorata (PO) were Diabetes mellitus (DM) was induced collected from Khon Kaen province of by a single dose of 55 mg/kg intraperitoneal Thailand. The fresh leaves of PO were dried (i.p.) streptozotocin (STZ) dissolved in 0.1 at 40 oC for 24 hours. The dried plants M sodium citrate buffer, pH 4.5. Then, blood materials were crushed and boiled in glucose levels (BGL) were obtained via tail distilled water for 30 minutes, filtered clip under anesthesia 3 days after STZ in- through nylon cloth and made to crude jection by using Accu-Check Performa test powder by using a lyophilizer. strips and meter. The BGL that higher than 2.2 Animals 250 mg/dl were considered as diabetic rats Male Wistar albino rats weighing and used for this study.1 180-220 grams were employed in this study. 2.4 Induction of peripheral neuropathy They were housed at the Animal Care Unit Peripheral neuropathy was induced of Faculty of Medicine, Khon Kaen by chronic constriction injury (CCI) after University, exposed to 12 hours light and induced diabetes.22 Each rat was anesthe- dark cycles, maintained in the animal tized with thiopental sodium (i.p.) 60 mg/ cages with free access to water and kg BW. At first, the hair of the rat’s lower standard laboratory diet (Charoen Pokphand back in the thigh region of right paw was Foods Public Company, Thailand). The shaved, and the skin was sterilized with experimental protocol was approved by the povidone-iodine. The skin of lateral surface Institutional Animal Ethics Committee of of right thigh was incised and sciatic nerve Khon Kaen University (AEKKU 16/2557). was exposed. Four ligations of 4-0 silk su- ture were ligated at proximal to the sciatic 2.3 Induction of diabetes mellitus trifurcation with about 1-mm spacing. Fi- STZ is a broad-spectrum antibiotic nally, skin was immediately sutured with extracted from Streptomyces acromogenes. 4-0 silk suture. The diabetogenic action of STZ was found 2.5 Experimental design to be a selective destruction of pancreatic The rats were divided into 9 groups beta cells and has been widely used to of 6 animals in each group as follow; 78 Vol.11 No.4 Group 1 (Normal control): Rats were Group 9 (DM + CCI + PO 120 mg/kg BW): not subjected to any surgical procedure. Diabetic rats with CCI model were adminis- Group 2 (DM + Vehicle): STZ-induced tered PO leaves extract dose 120 mg/kg BW. diabetic control rats were subjected and All rats were administered per oral distilled water was administered. (p.o.) for 21 consecutive days that starting Group 3 (DM + CCI + Vehicle): Dia- from the 1st day. During experiment, the betic rats were subjected to surgical proce- behavioral examinations including hot dure, ligated the sciatic nerve by CCI model plate, Von Frey hair, and sciatic functional and distilled water was administered. index tests were measured on day 1, 7, 14 Group 4 (DM + Sham + Vehicle): Dia- and 21 whereas nerve conduction velocity betic rats were subjected to surgical proce- test was measured on day 21 after CCI dure to expose the sciatic nerve without any procedure. ligation and distilled water was administered. 2.6 Behavioral examinations Group 5 (DM + CCI + Ascorbic acid): 2.6.1 Hot plate test Diabetic rats were subjected to a surgical Hot plate test was used to inves- procedure to ligate the sciatic nerve by CCI tigate sensory functional recovery. The rats model and ascorbic acid 100 mg/kg BW was were placed on a hot-plate surface with the administered. temperature adjust to 55 + 1 oC. The latency Group 6 (DM + CCI + Gabapentin): of the first sign of paw licking or jumping Diabetic rats were subjected to surgical (withdrawal response) to avoid the heat was procedure to ligate the sciatic nerve by CCI taken as an index of the pain threshold. The model and gabapentin 50 mg/kg BW was cut-off time of 10 seconds was maintained administered. to avoid damage to the paw.10 Group 7 (DM + CCI + PO 3 mg/kg BW): 2.6.2 Von Frey hair test Diabetic rats with CCI model were adminis- Von Frey hair test used for assess tered PO leaves extract dose 3 mg/kg BW. mechanical withdrawal threshold. Each rat Group 8 (DM + CCI + PO 30 mg/kg BW): was placed in a chamber with a wire grid Diabetic rats with CCI model were adminis- and the central region of the plantar surface tered PO leaves extract dose 30 mg/kg BW. of the hind paw was stimulated with a 79 Vol.11 No.4 series of ascending force von Frey mono- (ii) toe spread factor (TSF) = (ETS- filaments. A trial of each monofilament was NTS)/NTS; applied 10 times that in increasing force (iii) intermediary toe spread factor until the rat withdraw the paw. The with- (ITF) = (EIT-NIT)/NIT. drawal threshold was taken as the lowest After that, these factors were then in- force that cause at least 5 times out of the corporated into the Bain-Mackinnon-Hunter 10 stimuli.21,23 (BMH) sciatic function index-formula26: 2.6.3 Sciatic functional index SFI = - 38.3 x PLF + 109.5 x TSF + 13.3 The sciatic functional index was x ITF - 8. used to assess the motor functional recovery 2.7 Nerve conduction velocity (NCV) following experimentally-induced injury in The NCV test is a measurement of the the sciatic nerve.24,25 The trials were done in speed of conduction of an electrical impulse a wooden walking track darkened at one end through a nerve that can determine nerve and covered with a sheet of white paper.
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