GC-MS analysis of essential oil extracted from Chimonanthus praecox flowers and exploration of its anti-inflammatory effect / 西安交通大学学报(医学版)

【Objective】 To clarify the composition of essential oil extracted from Chimonanthus praecox flowers in Shiyan area and to explore its anti-inflammatory effects to provide support for further development of the resources of Chimonanthus praecox flowers. 【Methods】 SD rats were randomly divided into blank control group, model group, positive control group (aspirin 200 mg/kg), and low-, medium- and high-dose essential oil groups (0.1, 0.3 and 0.8g/kg). The blank control group and the model group were treated with distilled water by intragastric administration. The positive control group was treated with aspirin by intragastric administration. The low, medium, and high doses of the essential oil extracted from Chimonanthus praecox flowers were given 10, 30 and 80 mL/L at 10 mL/kg once a day. On day 5 of the experiment, 30 minutes after intragastric administration, 0.1 mL of Freund’s complete adjuvant was injected subcutaneously into the right foot plantar of each group of rats, and the blank control group was subcutaneously injected with 0.1 mL of normal saline. We observed and measured the toe’s volume of the rats before and 1, 2, 3, 5, 24, 48, and 72 h after injection by using drainage method. We then calculated the toe’s swelling rate in each group of rats at each time point, and used ELISA kit to measure the content of inflammatory factors in swollen foot tissue. 【Results】 In the medium- and high-dose essential oil groups, we observed significant inhibitory effects on the toe’s swelling rate in rats at 1, 2, 3, 5, 24, and 48 h after inflammation with Freund’s complete adjuvant (P<0.05). The essential oil extracted from Chimonanthus praecox flowers could significantly decrease the contents of TNF-α and IL-1β in the swollen foot tissue, and its anti-inflammatory effect was dose-dependent. 【Conclusion】 The essential oil extracted from Chimonanthus praecox flowers has obvious inhibitory effects on the rate of the toe’s swelling induced by Freund’s complete adjuvant. The anti-inflammatory effect may be related to the inhibition of TNF-α and IL-1β, but its anti-inflammatory effect is weaker than that of aspirin.

Emerging intervention of antidepressant with DMARD in non-cancerous nociceptive persistent pain associated depression in FCA induced rheumatoid arthritic rats

Rheumatoid arthritis (RA) is a chronic inflammatory disorder that causes pain, systemic complications and premature mortality. Depression has also been identified as a problem for persons with RA. This association remaining significant even after the degree of disease activity is controlled. In the present study, the efficacy of combination therapeutic effect of antidepressant (amitriptyline) with Disease Modifying Anti rheumatoid drug (leflunomide) was determined in rheumatoid arthritis pain associated depression in Freund's complete adjuvant (FCA) induced arthritic rats. Drug treatment was started 9 days after induction of FCA induced arthritis in rats. The antiarthritic activity was assessed by measuring paw volume, weight-bearing, hematological, biochemical, serological parameters, Radiographic analysis and Histopathology of tibiotarsal joints. The antidepressant activity was assessed by Forced swimming test, Rota-rod test and confirmed by estimation of brain neuro transmitters (serotonin and norepinephrine) level. Results of this study revealed that leflunomide and amitriptyline combination showed more significant (p<0.001) antiarthritic and antidepressant action and leflunomide alone treatment showed significant (p<0.001) antiarthritic activity only as compared to arthritic control. The leflunomide and low dose amitriptyline combination found to be more effective in pain associated depression in rheumatoid arthritic rats

Synthesis And Evaluation Of Phthalate Analogue Of Diclofenac Against Freund’s Complete Adjuvant Induced Arthritis In Rat

Objective: The objective of the present study is to evaluate the effect of Phthalate analogues of diclofenac in Freund’s complete adjuvant (FCA) induced Arthritis in the rat. Methods: Twenty four female albino wistar rats were enrolled in this study and are divided into 4 groups (six each). The groups were designed as follows: Group I: vehicle control, Group II: arthritic control, Group III: diclofenac treated, Group IV: phthalate analogue of diclofenac treated. Various assessments such as anti-arthritic activity, biochemical estimations, haematological parameters, ulcerogenesis, radiological and histopathological studies were evaluated. Results: Arthritic control group exhibited significant increase in the level of paw volume, arthritic score (p<0.0001), Serum glutamic pyruvic transaminase (SGPT) (p<0.001), Serum glutamic oxaloacetic transaminase (SGOT) p<0.01), rheumatoid arthritis factor, C-reactive protein (CRP), White Blood Cells (WBC), Creatinine and uric acid and a significant decrease in Red Blood Cells (RBC). Increased swelling of joints, bony destruction and profound ulceration were observed in the Arthritic control group. All these conditions were reversed in diclofenac and phthalate analogue of diclofenac groups. Conclusion: We conclude that phthalate analogue of diclofenac shows potent anti-arthritic activity with milder ulceration when compared to diclofenac treatment.

Evaluation of in-vivo antiarthritic potential of methanolic extract of Costus speciosus rhizome.

Objective: To evaluate anti-arthritic potential of methanolic extract of rhizome of Costus speciosus Koen. Methods: The powdered drug was extracted with 80% methanol. The crude extract was subjected to phytochemical investigation and was evaluated for its anti-arthritic potential by freund’s complete adjuvant induced arthritis model in adult wistar albino rats. Determination of different parameters like arthritic score, arthritic index, paw thickness, body weight, and pain, altered liver enzymes and biochemical estimation like, nitric oxide level and Plasma TNF-α level was done. Finally, radiological estimation and histopathology of tibio tarsal joints was performed. Statistical analysis was performed using one way ANOVA followed by Dunnet’s test at different p-values. Results: Phytochemical study revealed the presence of flavonoids, phenolic compound, saponins and carbohydrates. For different parameter mentioned above, anti-arthritic activity shown by prophylactic high dose test extract (200 mg/kg) was as potent as standard drug Indomethacin (10 mg/kg). The effect of Prophylactic low dose extract (100 mg/kg) and therapeutic high dose extract (200 mg/kg) was less than that of Indomethacin (10 mg/kg). Furthermore, Therapeutic low dose extract (100 mg/kg) was not effective. Conclusions: The obtained results indicate that Costus speciosus rhizome extract possess significant anti-arthrtic potential.

Roles of BCG in establishing collagen-induced arthritis mice / 中国免疫学杂志

Objective:To determine the correlation of BCG concentration in Freund complete adjuvant ( FCA) and severity of arthritis during arthritis establishment in DBA 1/j male mice induced by bovine type Ⅱcollagen.Methods:CIA was induced by the im-munization of DBA1/j mice with bovine type Ⅱcollagen and BCG of various concentrations dissolved in FCA.To ascertain the effects administering the collagen booster,CIA-related features(including body weight,clinical scoring of arthritis),TNF-αin serum and the histopathological changes in the spleen and the joints regions were measured.Results:4 mg/ml BCG induced more serious arthritis than 1 mg/ml in DBA1/j mice after collagen exposure.Conclusion:There is a positive correlation of arthritis severity with BCG.Which indicates that selection of adjuvant with suitable BCG concentration could determine pathological outcome in CIA mouse model .

Evaluation of anti-arthritic potential of the methanolic extract of the aerial parts of Costus speciosus.

Objective: Costus speciosus Koen. (Keu, Crape ginger), an ornamental plant, widely distributed in India is traditionally used as astringent, aphrodisiac, purgative, anthelmintic, depurative, febrifuge and expectorant. The plant is also used in rheumatism, dropsy, urinary diseases and jaundice. The purpose of this study is to evaluate the anti-arthritic activity of the methanolic extract of the aerial parts of Costus speciosus (CS) in experimental animal models. Materials and Methods: The powdered drug was subjected to successive solvent extraction, with solvents in increasing order of polarity to obtain the methanolic extract of the aerial parts of the plant. CS was evaluated for anti-arthritic action by Freund’s adjuvant induced arthritis test in adult Albino rats (150-200 gm). Rats were injected 0.1 ml of complete Freund’s adjuvant into the planter region of the left hind paw. Statistical analysis was performed using One way analysis of variance (ANOVA) followed by Bonferonni test. P<0.05 was considered statistically significant. Results: The methanolic extract of CS in doses of 400 and 800 mg/kg showed 75.50% and 68.33% protection against increase in paw edema, respectively. CS showed dose-dependent action in all the experimental models. Conclusion: The present study indicates that CS has significant anti-arthritic properties.

Comparison of Antiallodynic Effects between Intrathecal Cholinesterase Inhibitors and NMDA Antagonists on Two Neuropathic Pain Rat Models / 대한마취과학회지

BACKGOUND: Cholinesterase inhibitors and N-methyl-D-aspartate (NMDA) antagonists reduce the mechanical allodynia in neuropathic pain models. In this study our aim was to compare the antiallodynic effects between intrathecal cholinesterase inhibitors and NMDA antagonists on two well-characterized neuropathic pain rat models. METHODS: Male Sprague Dawley rats were anesthetized and either had the left L5 and L6 spinal nerves ligated (SNL group) or Freund complete adjuvant (FCA) administrated to the sciatic nerve (FCA group) in order to cause neuropathic pain. A catheter was implanted into the intrathecal space for drug administration. After obtaining baseline values, edrophonium (3-100microgram), neostigmine (0.3-10microgram), AP-5 (0.3-3microgram) and MK-801 (1-30microgram) were administered intrathecally to each group. The allodynic left hind paw withdrawal thresholds to von Frey hairs were assessed and converted to % MPE. Antiallodynic effects on the two groups were compared by analyzing dose-response curves and ED 50 values. Motor weakness was also checked. RESULTS: Intrathecal edrophonium, neostigmine, AP-5 and MK-801 had a dose-dependent antiallodynic effect on the two neuropathic pain models. Comparing the antiallodynic effect dose response curves, intrathecal cholinesterase inhibitors had lower ED 50 with steep slopes in the SNL model, whereas intrathecal NMDA antagonists had lower ED 50 in the FCA model, but there were no statistically significant differences between the two models. CONCLUSIONS: Intrathecal cholinesterase inhibitors and NMDA antagonists have relatively better antiallodynic effects on the SNL and FCA neuropathic pain rat models, respectively.

The Effect of alpha2 Adrenergic Agonists and Norepinephrine on Mechanical Allodynia by Freund's Complete Adjuvant Induced Inflammation in Rats / 대한마취과학회지

BACKGROUND: The Freund's complete adjuvant (FCA)-induced inflammation may produce allodynia against a touch stimulus. The antiallodynic effects of brimonidine, a new selective alpha2 receptor agonist, and of rilmenidine, a new more selective imidazoline receptor agonist, have not been evaluated in rats with FCA induced inflammation. Therefore, we investigated the sympathetic component of mechanical allodynia after the development of allodynia secondary to FCA-induced inflammation in rats. METHODS: A lumbar intrathecal catheter was implantated in male Sprague Dawley rats. Inflammation was induced by the intradermal injection of 0.15 ml FCA under enflurane anesthesia. Using Von Frey filaments, the antiallodynic effects of intrathecal (I.T.) brimonidine (1, 3 microgram), rilmenidine (30, 100 microgram) and saline were examined. In antagonistic study intrathecal yohimbine 30 microgram and rauwolscine 30 microgram were administered to investigate the reversal of the antiallodynic effect by each agonist. We also examined the effects of intradermal norepinephrine followed by I.T. brimonidin, rilmenidine or saline on the withdrawal threshold of rats secondary to allodynia induced by FCA. RESULTS: I.T. brimonidine or rilmenidine produced dose-dependent antiallodynic effect and which were moderately antagonized by I.T. yohimbine or rauwolscine. Intradermal norepinephrine produced a reduction in the withdrawal threshold in rats. CONCLUSIONS: Our results suggest that a sympathetic component is likely to be involved in the mechanism of allodynia secondary to FCA-induced inflammation.

Effect of Adrenal Gland Transplantation on Mechanical Hyperalgesia Induced by Freund's Complete Adjuvant in Rats / 대한마취과학회지

BACKGROUND: The transplantation of adrenal medullary tissue into the CNS of the rat can reduce pain. This results from the release of opioid peptides and catecholamines from the transplanted chromaffin cells. However, whether the adrenal gland transplanted in peripheral tissue can also reduce the pain is not well documented. The purpose of this study was to assess the potential for theadrenal gland implanted into the peripheral tissue of the rat to reduce mechanical hyperalgesia induced by inflammation. METHODS: Fifteen male Wistar rats were divided into three groups; one for implantation of the adrenal gland (AG), another for the kidney (Kd), and the other for a Sham operation (S). Tissues for transplantation were harvested from Sprague-Dawley rats. Two weeks after surgery, inflammation was induced by injecting 0.05 ml of Freund's Complete Adjuvant (FCA) into the dorsum of the rat's hind paw. Mechanical hyperalgesia was assessed using automated Randall-Sellito algesiometer at 6 hr, 1, 2, 3, 7 and 14 days after injecting FCA. RESULTS: Paw withdrawal thresholds against mechanical stimuli were 84.8 6.9%, 112.1 4.6%, 86.3 8.5%, 93.6 7.4%, 90.5 3.8%, 96.9 3.8%, respectively, in the AG group, and 71.3 10.8%, 93.3 11.3%, 79.1 10.3%, 73.7 8.1%, 62.6 6.4%, 77.7 6.2%, respectively, in Kd group, and 67.2 8.3%, 88.1 11.7%, 67.7 8.1%, 69.6 8.2%, 74.5 8.5%, 81.2 6.6%, respectively, in the S group. The AG group showed less pain sensitivity compared with that of the Kd and S groups especially 7 and 14 days after injecting the drug. CONCLUSIONS: This study indicates that the transplanted adrenal gland into the peripheral tissue may provide analgesic effect for a long time after it is transplanted in the rat.

Effect of Adrenal Gland Transplantation on Mechanical Hyperalgesia Induced by Freund's Complete Adjuvant in Rats / 대한마취과학회지

BACKGROUND: The transplantation of adrenal medullary tissue into the CNS of the rat can reduce pain. This results from the release of opioid peptides and catecholamines from the transplanted chromaffin cells. However, whether the adrenal gland transplanted in peripheral tissue can also reduce the pain is not well documented. The purpose of this study was to assess the potential for theadrenal gland implanted into the peripheral tissue of the rat to reduce mechanical hyperalgesia induced by inflammation. METHODS: Fifteen male Wistar rats were divided into three groups; one for implantation of the adrenal gland (AG), another for the kidney (Kd), and the other for a Sham operation (S). Tissues for transplantation were harvested from Sprague-Dawley rats. Two weeks after surgery, inflammation was induced by injecting 0.05 ml of Freund's Complete Adjuvant (FCA) into the dorsum of the rat's hind paw. Mechanical hyperalgesia was assessed using automated Randall-Sellito algesiometer at 6 hr, 1, 2, 3, 7 and 14 days after injecting FCA. RESULTS: Paw withdrawal thresholds against mechanical stimuli were 84.8 6.9%, 112.1 4.6%, 86.3 8.5%, 93.6 7.4%, 90.5 3.8%, 96.9 3.8%, respectively, in the AG group, and 71.3 10.8%, 93.3 11.3%, 79.1 10.3%, 73.7 8.1%, 62.6 6.4%, 77.7 6.2%, respectively, in Kd group, and 67.2 8.3%, 88.1 11.7%, 67.7 8.1%, 69.6 8.2%, 74.5 8.5%, 81.2 6.6%, respectively, in the S group. The AG group showed less pain sensitivity compared with that of the Kd and S groups especially 7 and 14 days after injecting the drug. CONCLUSIONS: This study indicates that the transplanted adrenal gland into the peripheral tissue may provide analgesic effect for a long time after it is transplanted in the rat.

Effects of Substance P Antagonist, D-Arg, D-Phe, D-Trp, Leu- Substance P on the Mechanical Hyperalgesia of Inflammation Induced by Freund's Complete Adjuvant in Rat Paw / 대한마취과학회지

BACKGROUND: The effect of substance P (SP) on the hyperalgesia induced by inflammation is controversial, and as SP remains in the periphery just for a short period of time after release from the nerve ending, the contribution of SP on the development of sustained mechanical hyperalgesia in rats with inflammation is questionable. The purpose of this experiment is to evaluate the effect of SP on the development of mechanical hyperalgesia induced by Freund's complete adjuvant (FCA) using SP antagonist [D-Arg, D-Phe, D-Trp, Leu]-substance P (SPA). METHODS: Male Sprague Dawley rats were divided into four groups; control (normal saline) and three different doses of SPA (0.25 microgram, 2.5 microgram, 25 microgram/0.1 ml). Inflammation was induced in rats by injecting 0.15 ml of FCA intraplantarly. Rats showed typical hyperalgesia within 12 hours after injection and maintained it for about one week. To test the effect of SPA on the developement of inflammation, either SPA or saline was injected at 1 h before and at the time of FCA injection under light halothane anesthesia after a baseline test. The effect of SPA on hyperalgesia was assessed by measuring mechanical hyperalgesia at 2, 6, 12, 24 hrs and 4 days after injection of the drug. To test the effect of SPA on fully developed inflammation, tests were done 2 days after injection of FCA. Mechanical hyperalgesias were assessed at 15, 30, 60, 90, 120 min after the drug injections. RESULTS: SPA injected to suppress the initial SP spill over decreased the mechanical hyperalgesia in a dose dependent manner. SPA injected after the full development of inflammation also decreased mechanical hyperalgesia. CONCLUSIONS: SP released at the initial phase of inflammation as well as SP released after the development of inflammation are all important for the maintainance of mechanical hyperalgesia.

The Role of Spinal NO on the Mechanical Allodynia in the Inflammation Model by Freund's Complete Adjuvant / 대한마취과학회지

BACKGROUND: The effect of spinal nitric oxide (NO) on mechanical allodynia brought about by Freund's complete adjuvant (FCA)-induced inflammation is not known. From our previous experiment nitric oxide synthase (NOS) inhibitor nitroG-L-arginine methyl ester (L-NAME) given intraplantarly during the induction period decreased a mechanical hyperalgesia occurring because of FCA-induced inflammation. Therefore, we investigated the spinal effect of NO on mechanical allodynia after the development of allodynia produced by FCA-induced inflammation in rats. METHODS: Male Sprague Dawley rats were prepared with lumbar intrathecal catheter implantation. Inflammation was induced in the rats by injecting 0.1 ml of FCA under halothane anesthesia. Behavioral tests were done 1, 3, 6, 24, and 48 hours after injection of FCA. In the other group, intrathecal L-NAME (10 microgram) was given prior to FCA injection to examine the effect of pretreatment. On postinjection day 2, either L-NAME (10 microgram) or methylene blue (10 and 30 microgram) was administered intrathecally after the baseline measurement. The withdrawal response on mechanical allodynia was assessed by applying von Frey filaments to the right lesioned hindpaw and contralateral paw (as control) at 15, 30, 45, 60, 90, and 120 minutes. Sodium nitroprusside was administered intrathecally to determine the reversal effect of increased threshold in the L-NAME group. RESULTS: Injection of FCA produced a significant mechanical allodynia over time. Pretreatment with L-NAME did not prevent such a mechanical allodynia. Intrathecal L-NAME, but not methylene blue, reduced the mechanical allodynia, which was reversed by sodium nitroprusside. CONCLUSIONS: Spinal NO is likely invloved in the mechanism of the development and maintenance of mechanical allodynia in a state of FCA-induced inflammation.