| The main symptoms of arthralgia syndrome are joint swelling and pain.Pain of arthralgia syndrome mostly belongs to chronic pain,ma nifested as hyperalgesia.The main symptom of arthralgia syndrome is joint swelling with chronic pain,manifested as hyperalgesia.Aconitum is bitter;bitter and sexually hot,which is the main medicine for treating cold-arthralgia syndrome.Aconitum,pungent,bitter andhot is an essential medicine for treating cold-arthralgia syndrome.The purpose of this study is to establish rat models of cold and heat arthralgia syndrome,and to observe the difference of analgesic effect of Aconitum on rats of cold and heat arthralgia syndrome,and to analyze its mechanism.thereby observing the difference of the analgesic effects exerted by Aconitum on the two different types of modelrats and analyzingthe underlying mechanisms.Part I:Study on rat models of cold and heat arthralgia syndromeObjective:To establish a rat model of cold-heat arthralgia syndrome and find out the difference of the effect of cold-dampness and dampness-heat factors on hyperalgesia of model rats.To establish rat models of cold and heat arthralgia syndrome and find out the different effects of the cold-dampness and heat-dampness factors on hyperalgesia in the model rats.Methods:CFA 0.1ml was subcutaneously injected into the toes of the right hind limb of ratsthe toes of the model rats'right hind limb,and the model rats and then the ratswere put into the artificial climate chamber box with good conditions and constant temperature for 4 hours each day.The environmental conditions of the model group of cold arthralgiaconditions for the cold arthralgia model group are as follows:the temperature is set to 10?,the humidity is 70%,and the wind speed is 1.5m/s;and for the heat-arthralgia model group,the temperature is set to30?,the humidity is 70%,and the wind speed is 1.5m/s.The 50%mechanical pain response value,pain response time and axillary temperatureof the rats were measured before and on the 3rd,5th,7th,9th,11th and 13th days a fter modeling,the body weightwere measured before and on the 3rd,7th and 14th days after modeling,and the changes of in diet,drinking water and urine volume were monitored 24 hours.The expression of 67inflammatory factors in L4-6 segment of spinal cord was detected by protein chip method.Results:The 50%mechanical pain response value and pain response time of model group rats decreased significantly on the 3rd day(compared with the baseline value P<0.01),and reached the peak value but peaked on the 7th and 9th day respectively,and lasted until the end of the experiment(compared with the control group P<0.01).After introducing cold and dampness factors,the sensitivity of the rats affected foot to mechanical stimulation in early ratsin th early stage decreased in varying degrees,and the effect was obvious on the 7th day(compared with model group P<0.01),but the 50%mechanical pain response value reduced with the prolongation of the modeling time in later stagein the later stage of modeling,and the effect was evidenton the 13th day(compared with model group P<0.05).After the introduction of cold factors,the increased sensitivity of affected foot to temperature stimulation could be inhibited to varying degrees in the early stage,especially on the 3rd and 7th days afte r modeling(P<0.01 compared with model group),but in the later stage,the sensitivity of affected foot to temperature stimulation could be increased to varying degrees with the prolongation of modeling,especially on the 9th and 11th days after modeling(P<0.05 compared with model group).There was no significant change in body temperature and dietary water intake of model rats after modeling,but after introducing cold-dampness and heat-dampness factors,the body weight and 24-hour total urination volume of model rats with cold-dampness and heat-dampness of cold and heat arthralgia model rats were different from those of control rats and model rats in varying degrees,manifesting in the decrease of body weight and urine volume,among which the introduction of cold-dampness factors played an obvious role.Fractalkine,a chemokine detected by protein chip,is a differentially expressed protein in line with the directionpurpose of this study.Conclusion:Fourteen days after subcutaneous injection of Freund's complete adjuvant into the toes of the right hind limb of rats,the affected feet of rats are sensitive to mechanical and temperature stimulation,and hyperalgesia is formedoccurs.The introduction of cold-dampness and heat-dampness factors had certain effects on the painalgesthesia of model rats,and cold-dampness factors could aggravate the hyperalgesia of model rats.The expression of Fractalkine protein in rat spinal cord can be used to analyze the effect of cold-dampness and dampness-heat on pain sensitivity in rats with arthralgia syndrome.Part ?:Study on theintervention effect of Aconitum on hyperalgesia in rats with cold-arthralgia and heat-arthralgiaObjective:To observe the difference of analgesic effect ofexerted by Aconitum on the model rats with cold arthralgia and heat arthralgia,and to analyze its mechanism.Methods:Rats were randomly divided into 9 groups:control group,model group,ibuprofen group,heat-arthralgia model group,cold-arthralgia model group,aconite-heat-arthralgia 1.6,0.8g crude drug/kg group,aconite-cold-arthralgia 1.6,0.8g crude drug/kg group.Except for the control group,the rats in each group were subcutaneously injected with CFA 0.1ml on the toes of the right hind limbs,and the model rats were put into the artificial climate box with good conditions and constant temperature for4 hours each day.The environmental conditions of for the model group of cold arthralgia are as follows:the temperature is set to 10? the humidity is 70%,the wind speed is 1.5m/s;and for the heat-arthralgia model group,the temperature is set to 30?,the humidity is 70%,and the wind speed is 1.5m/s.The 50%mechanical pain response value and pain response time of rats in each group were measured before and on the 3rd,7th,10th and 14th days after modelingrespectively.The expressions of CDllb/c,FNK,CX3CR1 and NMDA in L4-6 segment of spinal cord and dorsal root ganglion were detected by Western Blot method and immunohistochermistry.Results:After the establishment of the models,the 50%mechanical pain response value and pain response time of the model rats were significantly reduced(P<0.01 compared with the basic value of the same group).The 50%mechanical pain response value and pain response time of the model group rats at each time point were significantly lower than thoseof the control group(P<0.01),and the 50%mechanical pain response value reached the lowest on the 7th day after the model establishment,and did not recover significantly onsharply untilthe 14th day.On the 3rd day after the introduction of cold-dampness factor,the sensitivity of the affected foot to mechanical stimulation was significantly reduced(P<0.01 compared with the model group),but on the 10th and 14th days,the 50%mechanical pain response value of the affected foot was significantly lower than that of the model group(P<0.05 compared with the model group).And the sensitivity of the affected foot to mechanical stimulation had little change at each time point after the introduction of dampness-heat factor(P<0.05 compared with the model group).However,there were significant differences between the cold-arthralgia model group and the heat-arthralgia model group on the 7th and 10th days after modeling(P<0.01,P<0.05 compared with the heat-arthralgia model group).Aconitum dosage groups could increase the 50%mechanical pain response value in different degrees.Aconitum 1.6g/kg had significant effects on the 10th and 14th days after administration,and Aconitum0.8g/kg had significant effects on the 10th and 14th days after administration(P<0.01,P<0.05 compared with cold-arthralgia model group).Aconitum dosage groups could improve the sensitivity of heat-arthralgia model rats to mechanical stimulation in varying degrees,and the greater the dosage,the greater the effectthe higher the dosage is,the stronger effect Aconitum will exert.Obviously,the 50%mechanical pain response value decreased in varying degrees.Aconitum 1.6g/kg had a significant effect on the 3rd,7th and 14th days after administration(P<0.01 compared with the heat-induced arthralgia model group),and Aconitum 0.8g/kg had a significant effect on the 3rd day after administration(P<0.05 compared with the heat-induced arthralgia model group).There were also differences between the same dose of Aconitum heat-arthralgiamodelgroup and cold-arthralgia model group.Aconitum Aconitum 1.6g/kg could significantly increase 50%mechanical pain response in rats with cold arthralgia at different time points after administration.Aconitum0.8g/kg on the 3rd,7th and 14th days after administration,the 50%mechanical pain response value of Aconitum cold-arthralgia ratsincreased significantly(P<0.01,P<0.05 compared with the same dose of Aconitum heat-arthralgiagroup and cold-arthralgia model group).The introduction of cold factors could inhibit the increase of the sensitivitythe increasing sensitivityof the affected foot to temperature stimulation in varying degrees.The effect was obvious on the 3rd and7th days after modeling(P<0.01 compared with the model group);the introduction of damp-heat factors on the3rd day could prolong the pain response time of the model rats(P<0.05compared with the model group);the effect ofat other time points was not obvious;the reaction time of the affected foot in the cold-arthralgia model group was significantly longer than that in the heat-arthralgia model group on the 7th and 10th days after the establishment of the model(P<0.01 compared with the heat-arthralgia model group).Aconitum dosage groups could prolong the pain response time ofcold-arthralgiarats'affectedfeetin varying degrees.Aconitum dosage groups could prolong the pain response time of feet in different degrees,and Aconitum 1.6g/kg had obvious effect on7th day(P<0.05 compared with cold-arthralgia model group);Aconitum dosage groups could improve the sensitivity of heat-arthralgia model rats to temperature stimulation in varying deg rees,which showed that the pain response time was shortened in varying degrees,and Aconitum 1.6g/kg had the most obvious effect on 3rd,7th and 10th day.Significant(P<0.05,P<0.01)compared with heat arthralgia model group.On the 7th day after administration of 0.8g/kg aconite,the pain response time of aconite cold Bi group was significantly higher than that of aconite heat Bi group at the same dose(P<0.05 compared with the same dose Aconitum heat arthralgia group and cold arthralgia model group).Western Blot and Immunohistochemistry were applied to detect the expression of NMDAR,CD11b/c,FNK and CX3CR1 in the affected spinal cord and dorsal root ganglion of the model grouprats(compared with the control group,P<0.01).The expressions of NMDAR,CD11b/c,FNK and CX3CR1 in the spinal cord of the model group rats with cold and heat arthralgia syndrome were higher than those in the model group,and the heat-arthralgia model group waseven higher than the cold-arthralgia model group(P<0.01,compared with the model group;P<0.01,compared with the heat-arthralgia model group).Aconitum can significantly inhibit the expression of NMDAR,CDllb/c,FNK and CX3CR1 in the spinal cord and dorsal root ganglion of the affected side of cold-arthralgia model rats(P<0.05,P<0.01 compared with cold arthralgia model group),but has the opposite effect on the heat-arthralgia rats(P<0.01 compared with heat-arthralgia model group).Conclusion:The introduction of cold and heat factors can aggravate the pain sensitivity of arthralgia model rats.Aconitum has an obvious analgesic effect on cold arthralgia,but it can aggravate heat-arthralgiapain.This effect of Aconitum is related to its regulation of the expression of FKN,CX3CR1,CD11b/c and NMDAR in the dorsal horn of spinal cord and dorsal root ganglion of rats. |
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