| Objective:1.Preliminary study on the anti-inflammatory effect of alcohol extract and water extract of Cratoxylon ligustrinum（Spach）Blume.,and compare the difference of the effects of the two extracts;2.Observe the analgesic,anti-inflammatory effects of Qingniu Jiedu Decoction;3.Preliminary study the anti-gout arthritis effect and mechanism of Qingniu Jiedu Decoction.Methods:1.Anti-inflammatory effect of Cratoxylon ligustrinum（Spach）Blume.:Kunming mice were divided into（blank group）,model group,positive drug group(dexamethasone acetate 5·10-3 g/kg),low and high dose group of alcohol extract of Cratoxylon ligustrinum（Spach）Blume.（13 g/kg,26 g/kg）,low and high dose group of water extract of Cratoxylon ligustrinum（Spach）Blume.（13 g/kg,26 g/kg）.The experiment used the tests of dimethylbenzene-induced ear swelling,cotton ball-induced mouse granuloma and 1%carrageenan-induced paw edema to study on anti-inflammatory effects of Cratoxylon ligustrinum（Spach）Blume.The content of TNF-αin serum of mice with toe swelling induced by carrageenan was determined by ELISA.2.Analgesic and anti-inflammatory effects of Qingniu Jiedu Decoction:Kunming mice were divided into（blank group）,model group,positive drug group(aspirin enteric-coated tablets 5·10-3 g/kg or dexamethasone acetate 5·10-3 g/kg),Qingniu Jiedu Decoction low,medium and high dose groups（34.125g/kg,68.25g/kg,136.5 g/kg）,to observe on the analgesic effect of Qingniu Jiedu Decoction by method method acetic acid writhing method,and used the tests of dimethylbenzene-induced ear swelling,cotton ball-induced mouse granuloma and 1%carrageenan-induced paw edema to study on anti-inflammatory effects of Qingniu Jiedu Decoction.The levels of IL-1βin serum and PGE2 in inflammation tissue of toe swelling mice were determined by ELISA.3.Anti-gouty arthritis effect of Qingniu Jiedu Decoction:SD rats were divided into blank group,model group,positive drug group(Colchicine 0.5·10-3 g/kg),Qingniu Jiedu Decoction low,medium and high dose groups（23.625 g/kg、47.25 g/kg、94.5 g/kg）,the rat joint swelling was measured before modeling and at 6,12,24,48 h,and gait and arthritis index were evaluated after 24 h.At 48 h after modeling,the levels of leukocytes,neutrophils and lymphocytes in the blood were measured,comparing the number of kidney system and spleen system in each group.By observing HE staining sections of the ankle joints of rats,the pathological changes of the tissues were analyzed.The DNA sequences of NLRP3,ASC,and Caspase-1 in the synovial tissue of the ankle joint were quantitatively analyzed by Real-time PCR.Meanwhile,the protein expressions of NLRP3,ASC and caspase-1 were qualitatively and quantitatively analyzed by immunohistochemical analysis of the ankle joint.The serum level of IL-1β,IL-2,IL-4,IL-6,IL-10 and TNF-αwere detected by Luminex multi-factor detection technique.Results:1.Anti-inflammatory effect of Cratoxylon ligustrinum（Spach）Blume.（1）The high dose group of alcohol extract and water extract of Cratoxylon ligustrinum（Spach）Blume.could significantly alleviate the occurrence of ear swelling caused by xylene（P<0.05）;（2）The high and low dose groups of Cratoxylon ligustrinum（Spach）Blume alcohol extract and water extract all had significant inhibition on cotton granuloma in mice（P<0.01）;（3）The high and low dose groups of Cratoxylon ligustrinum（Spach）Blume aqueous extract and alcohol extract had significant inhibitory effect on mouse toe swelling（P<0.05,P<0.01）.2.Analgesic and anti-inflammatory effects of Qingniu Jiedu Decoction（1）The medium and high dose group of Qingniu Jiedu Decoction could significantly reduce the number of writhing in mice（P<0.05,P<0.01）;（2）The Qingniu Jiedu Decoction could significantly reduce the occurrence of xylene-induced ear swelling in mice（P<0.05）;（3）The Qingniu Jiedu Decoction had a significant inhibitory effect on cotton granuloma in mice（P<0.01）;（4）The Qingniu Jiedu Decoction had a significant inhibitory effect on the swelling of the toes of mice（P<0.01）;（5）The Qingniujiedu decoction had significant inhibitory effects on PGE2 in the inflammatory tissue of swollen toe mice（P<0.05,P<0.01）.3.Anti-gouty arthritis effect of Qingniu Jiedu Decoction（1）After modeling,at 6 h,12h,24h and 48 h,the medium,high groups of Qingniu Jiedu Decoction could significantly inhibit the joint swelling of the ankle joint（P<0.05,P<0.01）;（2）Qingniu jiedu decoction could significantly improve the gait and the state of joint inflammation of rats with acute gouty arthritis 24h after the establishment of the model（P<0.05,P<0.01）.（3）Qingniu jiedu decoction could significantly inhibit the increase of white blood cells and neutrophils（P<0.01,P<0.05）,and its high-dose group could significantly inhibit the increase of lymphocytes（P<0.05）.（4）For the spleen coefficient,the difference between the high dose group of Qingniu Jiedu Decoction was statistically significant（P<0.01）.（5）For IL-1β,IL-10,the medium dose group of Qingniu Jiedu Decoction had significant difference（P<0.05）;For IL-4,there was a significant difference between the low and high dose groups of Qingniu Detoxification Decoction（P<0.01）;For IL-6,there was a significant difference in the high-dose group of Qingniu Jiedu Decoction（P<0.05）;（6）Qingniu jiedu decoction could significantly improve the inflammatory infiltration and edema of synovial tissue in rats with gout arthritis.（7）Qingniu Jiedu Decoction could significantly inhibit the m RNA expression of NLRP3（P<0.01,P<0.05）.The middle dose group of Qingniu Jiedu Decoction could significantly inhibit the m RNA expression of ASC（P<0.01）,and the high dose group of Qingniu Jiedu Decoction could significantly inhibit the m RNA expression of Caspase-1（P<0.01）.（8）The protein expression of NLRP3 was significantly inhibited in the medium and high dose group of qingniu jiedu decoction（P<0.01,P<0.05）,the protein expression of ASC was significantly inhibited in the low,medium and high dose group of qingniu jiedu decoction（P<0.01,P<0.05）,and the protein expression of caspase-1 was significantly inhibited in the qingniu jiedu decoction group（P<0.01,P<0.05）.Conclusion:1.Cratoxylon ligustrinum（Spach）Blume.has obvious anti-inflammatory effects about the tests of dimethylbenzene-induced ear swelling,1%carrageenan-induced paw edema and cotton ball-induced mouse granuloma;2.Qingniu Jiedu Decoction has a good inhibitory effect on the writhing reaction induced by acetic acid in mice,and the reaction of xylene-induced mouse ear swelling,cotton ball-induced mouse granuloma and carrageenan-induced toe swelling in mice good anti-inflammatory effect,while reducing the production of PGE2 inflammatory factors induced by carrageenan;3.Qingniu Jiedu Decoction has anti-gout arthritis effect,and can reduce the content of IL-1βin serum,increase the content of IL-4 and IL-10,inhibit the m RNA expression of NLRP3,ASC and Caspase-1,and regulate the NLRP3/ASC/Caspase-1 axis for the role of gouty arthritis. |
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