| SLW was recorded in《experienced effective prescription》early,composed of Sparganium stoloniferum Buch-Ham and Curcuma Wenchowensis Y.H.Chen et C. Ling which both can activate blood circulation to dissipate blood stasis. SLW was used to cure either stagnant blood or amenia,and stomachache corresponding with menst.The study of our topic group showed that SLW could deflate ectoped endomembrane and the serum containing sparganii and curcumae could inhibit endothelial cell proliferation of blood vessel induced by VEGF.EMS is a diease correlated with inflammation and pain.At present we don not know if SLW have a analgesic and anti?inflammatory effect.Our test study the analgesic and anti-inflammatory effects and mechanism of SLW to offer experiment foundation of the future new drug used to therapy EMS.Objective:To study the analgesic and anti-inflammatory effects and mechanism of Sanlingwan (SLW).Methods:1 Analgesic effect test 1.1 Hot plate test: Female mouse of Kunming stain whose Algesia response latency were between 10s～30s were Selected and were distributed to five groups. Mouse in SLW groups were given SLW (1.43,0.48,0.16 mg·kg-1, p. o) and mouse in Tramadol group were administered Tramadol (20mg·kg-1, p.o) and mouse in model group were administered physiologic saline(0.2 mL·10 g-1, p. o).Tirty minutes after p. o , each mice was put on hot plate of 55℃respectively to determine algesia response latency of each mice.1.2 Writhing test: Mouse of Kunming stain were Selected and were distributed to five groups. Mouse in SLW groups were given SLW (1.43,0.48,0.16 mg·kg-1, p. o,daily) and mouse in celecoxib group were administered celecoxib (25mg·kg-1, p.o,daily) and mouse in model group were administered physiologic saline(0.2 mL·10 g-1, p. o,daily) for three days. Tirty minutes after the late p. o ,each mice was given 0.7%HAc(0.1 mL·10 g-1,i.p) respectively to determine writhing times of each mice in 20 minutes.2 Anti-inflammatory effect test2.1 The permeability increase of enterocoelia blood capillary induced by HAc in mice: Mouse of Kunming stain were Selected and were distributed to five groups. Mouse in SLW groups were given SLW (1.43,0.48,0.16 mg·kg-1, p. o,daily) and mouse in celecoxib group were administered celecoxib (25mg·kg-1, p.o,daily) and mouse in model group and normal group were administered physiologic saline(0.2 mL·10 g-1, p. o,daily) for three days. Tirty minutes after the late p. o ,each mice was given 0.5% Evans Blue(0.1 mL·10 g-1,ip)and given 0.7% HAc immediately(0.1 mL·10 g-1,i.p)except normol group respectively to determine enterocoelia Evans Blue concentration of solution of each mice .2.2 Ear swelling test: Mouse of Kunming stain were Selected and were distributed to five groups. Mouse in SLW groups were given SLW (1.43,0.48,0.16 mg·kg-1, p. o,daily) and mouse in celecoxib group were administered celecoxib (25mg·kg-1, p.o,daily) and mouse in model group were administered physiologic saline(0.2 mL·10 g-1, p. o,daily) for three days. Tirty minutes after the late p. o ,each mice was painted by 100%xylene (0.01 mL) respectively to determine the swelling of ear of each mice.2.3 Paw edema test in rat: Male rats of Wistar stain were Selected and were distributed to five groups. Rats in SLW groups were given SLW (1.02,0.34,0.11 mg·kg-1, p. o,daily) and rats in celecoxib group were administered celecoxib (20mg·kg-1, p.o,daily) and rats in model group were administered physiologic saline(0.1 mL·10 g-1, p. o,daily) for three days. After the late p. o ,each rat was given 1%CAR (0.1 mL,sc)in light hind paw respectively to determine the edema of paw of each rat.2.4 The cotton?pellet granloma of the rat: Male rats of Wistar stain were Selected and were distributed to five groups. The cotton?pellets (30 mg±0.5 mg)were implanted into fold inguen of two sides subcutaneously. Rats in SLW groups were given SLW (1.02,0.34,0.11 mg·kg-1, p. o,daily) and rats in celecoxib group were administered celecoxib (20mg·kg-1, p.o,daily) and rats in model group were administered physiologic saline(0.1 mL·10 g-1, p. o,daily) for seven days. Each cotton?pellet was taken out in the eighth day respectively to determine granloma weight of each rat .3 Anti?inflammatory mechanism test3.1 The amounts of PGE2 in the carrageenin-induced inflammatory diffusate: Male rats of Wistar stain were Selected and were distributed to six groups. Rats in SLW groups were given SLW (1.02,0.34,0.11 mg·kg-1, p. o,daily) and rats in celecoxib group were administered celecoxib (20mg·kg-1, p.o,daily) and rats in model group and normal group were administered physiologic saline(0.1 mL·10 g-1, p. o,daily) for three days. After the late p. o ,each rat was given 1%CAR (0.1 mL,sc)in light hind paw respectively to determine the amounts of prostaglandin E2 (PGE2) in the carrageenin-induced inflammatory diffusate of each rat.3.2 The amounts of Hist in the carrageenin-induced inflammatory diffusate: Male rats of SD stain were Selected and were distributed to six groups. Rats in SLW groups were given SLW (1.02,0.34,0.11 mg·kg-1, p. o,daily) and rats in celecoxib group were administered celecoxib (20mg·kg-1, p.o,daily) and rats in model group and normal group were administered physiologic saline(0.1 mL·10 g-1, p. o,daily) for three days. Thirty minutes after p. o ,each rat was given 1%CAR (0.1 mL,sc)in light hind paw respectively to determine the amounts of Hist(histamine) and in the carrageenin-induced inflammatory diffusate of each rat.3.3 The amounts of 5-HT in the carrageenin-induced inflammatory diffusate: Male rats of SD stain were Selected and were distributed to six groups. Rats in SLW groups were given SLW (1.02,0.34,0.11 mg·kg-1, p. o,daily) and rats in celecoxib group were administered celecoxib (20mg·kg-1, p.o,daily) and rats in model group and normal group were administered physiologic saline(0.1 mL·10 g-1, p. o,daily) for three days. Thirty minutes after p. o ,each rat was given 1%CAR (0.1 mL,sc)in light hind paw respectively to determine the amounts of 5-HT(5-hydroxytryptamine) in the carrageenin-induced inflammatory diffusate of each rat.3.4 Ear swelling test in adrenalectomized mice: Adrenalectomized mouse of Kunming stain were selected and were distributed to five groups. Mouse in SLW groups were given SLW (1.43,0.48,0.16 mg·kg-1, p. o,daily) and mouse in celecoxib group were administered celecoxib (25mg·kg-1, p.o,daily) and mouse in model group were administered physiologic saline(0.2 mL·10 g-1, p. o,daily) for three days. Tirty minutes after the late p. o ,each mice was painted by 100%xylene (0.01 mL) respectively to determine the swelling of ear of each mice. Results:1 Analgesic effect test1.1 Hot plate test: Compared with the model group , a 1.43 mg·kg-1·d-1 dose of SLW could remarkably prolong the algesia response latency (P<0.01);either 0.48 mg·kg-1·d-1 or 0.16 mg·kg-1·d-1 dose of SLW could not remarkably prolong the algesia response latency (P>0.05). The result showed that a 1.43 mg·kg-1·d-1 dose of SLW could remarkably have analgesic effect.1.2 Writhing test: Compared with the model group , a 1.43 mg·kg-1·d-1 dose of SLW could remarkably prolong the writhing response latency (P<0.01);either 1.43 mg·kg-1·d-1 or 0.48 mg·kg-1·d-1 dose of SLW could remarkably decrease the writhing times(P<0.01). The result showed that SLW could remarkably reduce writhing response.2 Anti-inflammatory effect test2.1 The permeability increase of enterocoelia blood capillary induced by HAc in mice: Compared with the normal group , the permeability content of the model group have remarkable increase (P<0.01).Compared with the model group ,a 0.16 mg·kg-1·d-1 dose of SLW could remarkably reduce the permeability increase (P<0.05);either 1.43 mg·kg-1·d-1 or 0.48 mg·kg-1·d-1 dose dose of SLW could remarkably reduce the permeability increase (P<0.01). The result showed that SLW could remarkably reduce the permeability increase of enterocoelia blood capillary induced by HAc. 2.2 Ear swelling test: Compared with the model group ,a 0.48 mg·kg-1·d-1 dose of SLW could remarkably reduce ear swelling (P<0.01). The result showed that a 0.48 mg·kg-1·d-1 dose of SLW could remarkably reduce the ear swelling of mice.2.3 Paw edema test in rat: Compared with the model group ,a 0.11 mg·kg-1·d-1 dose of SLW could remarkably reduce paw edema (3h after inducing inflammation, P<0.01;4h after inducing inflammation, P<0.05); a 1.02 mg·kg-1·d-1 dose could remarkably reduce paw edema (2h and 3h after inducing inflammation, P<0.01;4h after inducing inflammation, P<0.05); a 0.34 mg·kg-1·d-1 dose could remarkably reduce paw edema (2h and 3h after inducing inflammation, P<0.01;1h and 4h after inducing inflammation, P<0.05). The result showed that SLW could remarkably reduce the paw edema in rat.2.4 The cotton?pellet granloma of the rat: Compared with the model group ,either 0.34 mg·kg-1·d-1 or 0.11 mg·kg-1·d-1 dose of SLW could remarkably reduce the granloma of the rat (P<0.05); a 1.02 mg·kg-1·d-1 dose could remarkably reduce the granloma (P<0.01). The result showed that SLW could remarkably reduce the granloma of the rat.3 Anti?inflammatory mechanism test3.1 The amounts of PGE2 in the carrageenin-induced inflammatory diffusate: Compared with the normal group , the amounts of PGE2 of the model group in the inflammatory diffusate have remarkable increase (P <0.01). Compared with the model group ,either 1.02 mg·kg-1·d-1 or 0.11 mg·kg-1·d-1 dose of SLW could remarkably reduce the amounts of PGE2 (P<0.05);a 0.34 mg·kg-1·d-1 dose could remarkably reduce the amounts of PGE2 (P<0.01). The result showed that SLW could remarkably reduce the amounts of PGE2.3.2 The amounts of Hist in the carrageenin-induced inflammatory diffusate: Compared with the normal group , the amounts of Hist of the model group in the inflammatory diffusate have remarkable increase (P<0.01). Compared with the model group , the respective dose of SLW could remarkably reduce the amounts of Hist (P<0.05). The result showed that SLW could remarkably reduce the amounts of Hist.3.3 The amounts of 5-HT in the carrageenin-induced inflammatory diffusate: Compared with the normal group , the amounts of 5-HT of the model group in the inflammatory diffusate have remarkable increase (P<0.01). Compared with the model group ,either 1.02 mg·kg-1·d-1 or 0.34 mg·kg-1·d-1 dose of SLW could remarkably reduce the amounts of 5-HT (P<0.01). The result showed that SLW could remarkably reduce the amounts of 5-HT.3.4 Ear swelling test in adrenalectomized mice: Compared with the model group ,a 0.48 mg·kg-1·d-1 dose of SLW could remarkably reduce ear swelling (P<0.05). The result showed that a 0.48 mg·kg-1·d-1 dose of SLW could remarkably reduce the ear swelling of mice and the anti-inflammatory effects of SLW were not correlated with adrenal.Conclusion:SLW had a significant analgesic and antiffinflammatory effect which may correlate with the inhibition of PGE2,Hist,5-HT. |
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