| OBJECTIVE: Qinjiao(Gentianae macrophyllae Radix),a mild-natured traditional Chinese medicine(TCM)of expelling wind-damp,was taken as a fundamental medicine,and matched up with Weilingxian(Clematidis Radix et Rhizoma),Sangjisheng(Taxilli Herba)and Fangji(Stephanlae Tetrandrae Radix)respectively,constituting a mild-warm compatibility(Qinjiao+Weilingxian),mild-mild compatibility(Qinjiao+Sangjisheng)and mild-cold compatibility(Qinjiao+Fangji),the relation between medicine property and disease and the relation between medicine property and curative effect were explored through an observation of the effect of different compatibilities of Qinjiao on wind-damp-heat arthralgia RA rats so as to interpret the correlation between TCM property,syndrome and curative effect,and to explore the mechanism of TCM treating RA,explore and establish the thought and method of studying the property of TCM based on traditional function.METHODS: Eighty Sprague-Dawley rats were randomly divided into the blank group,collage?model group,wind-damp-heat arthralgia model group,Tripterygium group,single Qinjiao group,Qinjiao-Weilingxian group(Qin-Wei group),Qinjiao-Sangjisheng group(Qin-Sang group),Qinjiao-Fangji group(Qin-Fang group),with 10 in each.Wind-Damp-Heat arthralgia model were induced by collage? and being exposed in wind-damp-heat environment:In the first day,except the blank group,the rats in the other groups were given 2 points(back spine on both sides),1 at the end of the tail,shaved and intradermally injected with 0.1mL C? emulsion for 0.3mL.Rats were injected with 0.1 m L of C? emulsion in the left hind paw of the rat to carry out secondary immunization on 12 th day.The blank group was injected with the same amount of saline at the same site.From the second day of the first injection of C? emulsion,except for the blank group and the C? group,the other groups were modeled wind-damp-heat arthralgia using the reconstructed artificial climate box.The wind-damp-heat environment conditions were as follows: temperature 36 ~ 38 ?,wind speed 5 m/s,relative humidity ?95%.In the experiment,the paw was soaked in 37 ? hot water,the rats were fumigated at 37 ?,30 minutes every day,for 18 consecutive days.After modeling,each administration group was gavaged with corresponding drug solution.and the rats in the blank group and collage?model group were given the same amount of normal saline for 21 days.During the experiment,the general state of the rats was observed and recorded.The body weight,the thickness of the paw were measured every 3 days,and calculate Swelling.The arthritis index(AI)was scored on a regular basis.The pain threshold,cold pain tolerance time and pressure pain tolerance,and paw temperature,rectal temperature was measured at 18 d,28 d and 38 d respectively.After the animals were sacrificed,2 rats were sacrificed in the left hind leg,fixed,decalcified,paraffin-embedded and HE stained.The pathological changes were observed under light microscope.The levels of serum tumor necrosis factor-?(TNF-?),interleukin-1?(IL-1?)and prostaglandin E2(PGE2)were measured by ELISA.Immunohisto-chemistry was used to detect the expression of matrix metalloproteinase-3(MMP-3),tissue metalloproteinase inhibitor-1(TIMP-1)in rat ankle joint,the expression was detected by semi-quantitative analysis.RESULTS:1 From the body weight,paw swelling,AI,compared with wind-damp-heat arthralgia model group,the paw swelling of treatment groups were significantly reduced(P<0.05 or P<0.01),the Qin-Fang group were lower most significant(P<0.01),and significantly lower than the Qin-Sang group(P<0.05);the AI of Qin-Fang group,single Qinjiao group,Qin-Sang group were significantly reduced(P<0.05 or P<0.01),the AI of Qin-Fang group was lowest,and significantly lower than Tripterygium group and Qin-Wei group(P<0.05).There was no significant difference in body weight during eight groups.2 From pain test and temperature test,compared with the wind-damp-heat arthralgia model group,the pain threshold of Qinjiao compatibility groups was significantly rised(P<0.01);cold pain tolerance time of treatment groups were significantly shorter(P<0.05 or P<0.01),the reduction of Qin-Fang group was most significant;the pressure pain tolerance of treatment groups were significantly increased(P<0.01),in which the Qin-Fang group was significantly increased than Tripterygium group,Qin-Sang group,and single Qinjiao group(P<0.01).The paw temperature of single Qinjiao group Qin-Fang group,Tripterygium group were significantly lower than wind-damp-heat arthralgia model group,Qin-Wei group,Qin-Sang group(P<0.05 or P<0.01).There was no significant difference in rectal temperature during eight groups.3 HE staining results showed,there were obvious articular surface damages,a large number of inflammatory cell infiltration and granulation tissue hyperplasia in collage?model group and wind-damp-heat arthralgia model group.Wind-damp-heat arthralgia model group got worse,and existed a wealth of new blood vessels.inflammatory cell infiltration,granulation tissue hyperplasia,and vascular proliferation of ankle in treatment group were reduced,the scar tissue increased,in which Qin-Fang group improved better,Qin-Sang group was poor.4 Serological test results shows,compared with the wind-damp-heat arthralgia model group,the content of TNF-? in the serum of Qin-Fang group was significantly lowest(P<0.01),followed by Tripterygium group,the Qin-Sang group,the Qin-Wei group(P<0.05),and the Qin-Fang group was significantly lower than single Qinjiao group(P<0.05);the content of IL-1? in the serum of Qin-Fang group was significantly lowest(P<0.01),followed by other groups(P<0.05),and the Qin-Fang group was significantly lower than Tripterygium group,single Qinjiao group,the Qin-Sang group(P<0.05 or P<0.01);the content of PGE2 in the serum of the Qin-Wei group,Qin-Fang group,single Qinjiao group were significantly lower(P<0.01),followed by the Qin-Sang group(P<0.05),the content of PGE2 in the serum of treatment groups were no significant difference5 Immunohistochemistry showed,compared with wind-damp-heat arthralgia model group,the expression of MMP-3 in all treatment groups were significantly lower(P<0.01),especially in the Qin-Fang group,the MMP-3 of Qin-Fang group was significantly lower than other administration groups;the expression of TIMP-1 were increased in varying degrees in all administration groups,in which the Tripterygium group,the Qin-Fang group,the Qin-Wei group were significantly increased(P<0.05 or P<0.01),and the expression of TIMP-1 in Qin-Fang group and Qin-Wei group were significantly increased than Tripterygium group,single Qinjiao group and the Qin-Sang group(P<0.05 or P<0.01).CONCLUSION:1 The model effect of wind-damp-heat arthralgia model we established is better than CIA,its condition is more serious and course is longer.To a certain extent,this model can simulate the features of human rheumatism.2 For the wind-damp-heat arthralgia RA,different compatibility of Qinjiao and single Qinjiao all have some treatment effect,the combined effect of mild and cold traditional Chinese medicine(TCM)is outstanding,better than mild-warm,mild-mild,and single.Experimental results are consistent with the principles of traditional Chinese medicine clinical treatment of "treating the hot diseases should use the cold medicine ".the mechanism of this compatibility drug relieving wind-damp-heat arthralgia RA may be related to its ability,which can reduce the content of TNF-??IL-1?PGE2 and the expression of MMP-3,increase the expression of TIMP-1,reduce articular cartilage destruction,and reduce inflammatory cell infiltration and vascular proliferation.3 From the scope of this study,TCM on the role of RA is comprehensive,comprehend-sive,multilayered,which not only can play the effect of anti-inflammatory analgesic,relieve RA,but also relieve or prevent the erosion and destruction of RA joints,repair damaged tissue,its complete mechanism needs further study. |
PDF Full Text Request