| Osteoarthritis is a degenerative disease of articular cartilage.OA incidence is 15%,at present,NSAIDs are widely used to treat OA,which can relieve pain,but the toxic side effects were high,and not long-term to use.Due to the composition synergy of Traditional Chinese Medicine,it can relieve symptoms and improve governance to achieve the effect of functionality.Transdermal local medication can avoid the first effect,which make the drug arrive the pathological parts directly,with lasting efficacy and stability,little toxic side effects.This research is based on the clinical practice of Xiongnai prescription in the treatment of OA,which developing into a thermosensitive gel preparation for external use.Objective:To provide scientific basis for Xiongnai prescription,this research will screen the effective parts of anti-osteoarthritis,optimize the preparation process and carry out quality investigation,study the preliminary pharmacodynamics and skin irritation.Methods:1.To establish the analysis methods of Xiongnai extract,and determine the effective part of Xiongnai on anti-osteoarthritis by using the anti-inflammatory and analgesic experiments and rat model of papain-induced osteoarthritis and.2.To optimize and produce Xiongnai thermosensitive gels by using Box-Behnken Design-Response Surface methodology and Z-Comprehensive evaluation method,and determine its drug release model in vitro.3.To investigate the physical and chemical properties of Xiongnai thermosensitive gels.TLC methods and HPLC methods were used to investigate qualitative identification and content determination,and inspect the stability of Xiongnai thermosensitive gels.4.To investigate the anti-osteoarthritis effect and skin irritation by using anti-inflammatory and analgesic experiments,anti-osteoarthritis experiments and New Zealand rabbits skin irritation experiments.Results:1.The average content of FA and EPMC in the alcohol extract were 0.787 mg·g-1 and 8.144 mg·g-1,and in the water extract were 0.607 mg·g-1 and 4.157 mg·g-1.Compared with the blank group,the ear swelling degree and twisting level of positive control group,Xiongnai water extract and alcohol extract group were obviously decreased(P<0.01),and the pain threshold increased percentage of Xiongnai alcohol extract group and Xiongnai water extract high dose group were significantly increased(P<0.01).Besides,the inhibition rate of ear swelling degree,twisting level and pain threshold increasing percentage of Xiongnai alcohol extract were higher than the water extract in the same dose.Compared with the model group,the ankle swelling,toes swelling and the content of IL-1β and NO in serum of EAE group and NBAE group were decreased significantly(P<0.01)after dosing two weeks,and EAE group better than NBAE(P<0.05).According to the pathological section of ankle,there were some inflammatory cells in the left ankle synovial tissue and the articular cartilage damaged no obvious in EAE group and NBAE group.2.The optimal prescription of Xiongnai thermosensitive gels was 18.04% poloxamer407,0.57% poloxamer188,1.25% azone:peppermint oil(1:1),the cold cut temperature was 3.96 ℃,and the normalized index prediction was 4.605.Acorrding to validation test,the gel temperature was 32.4 ℃,viscosity was 15.2 Pa·s,and the accumulated transdermal flux(12h)was 144.848 μg·cm-2 in average.FA release was conformed to zero order equation,and the penetration constant was 0.9910.3.Xiongnai thermosensitive gels was dark brown,exquisite and no mass,and the average pH was 5.66,the average viscosity was 15.20 Pa·s,the average gel temperature was 32.2 ℃.According to the TLC,FA and EPMC in Xiongnai thermosensitive gels revealed the bright spot in the corresponding positions.The average content of FA and EPMC in Xiongnai thermosensitive gels was 7.6955 mg·g-1 and 2.5440 mg·g-1.Xiongnai thermosensitive gels was uniform,but not hierarchical after centrifugation at 3000r·min-1.There was no hierarchical and obvious changes in appearance,content of FA and EPMC in conditions of(25±2)℃,(40±2)℃,the humidity of(90±5)%,the illumination of(4500±500)lx and avoid light for six months at(4±2)℃.In conditions of(60±2)℃,the content of FA was decreased,and in conditions of the humidity of(75±5)%,the content of FA and EPMC were increased.4.Compared with the blank group,the level of ear swelling degree,absorbance and twisting were significantly decreased in Xiongnai thermosensitive gels group(P<0.05),and the level of pain threshold increasing percentage was obviously increased(P<0.05).Compared with the model group,the ankle swelling,toes swelling and the content of IL-1β and NO in serum were decreased(P<0.05)in Xiongnai thermosensitive gels group.According to pathological section,there were little of inflammatory cells in the left ankle synovial tissue and the articular cartilage damaged no obvious in Xiongnai thermosensitive gels group.The skin irritation integral in single time was 0,and in multiple times was 0.45 after removing Xiongnai thermosensitive gels.According to pathological section,the cutin cell layer was complete and continuous,and there was no inflammatory cells,and there was no different structure and organization in Xiongnai thermosensitive gels group.Conclusion:The content of FA and EPMC in Xiongnai alcohol extract are higher than the water extract,and the effect of anti-inflammatory and analgesic in Xiongnai alcohol extract is stronger than the water extract.The ethyl acetate extract of Xiongnai alcohol extract is the most effective part on anti-osteoarthritis.Xiongnai thermosensitive gels is good formability,uniform appearance and stable quality.The quality control method of Xiongnai thermosensitive gels is accurate,reliable and strong specificity.Xiongnai thermosensitive gels has the effect on anti-osteoarthritis and no skin irritation. |
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