| Objective:Three differently directional branches of Cinnamomi Ramulus and Cinnamomi Cortex were selected,and comparative study on their content of Cinnamaldehyde,chemical constituents of the volatile oil components and anti-inflammatory effect on adjuvant arthritis in rats were used,in order to evaluate their components and pharmacodynamic difference,objective to explore the diameter range of Cinnamomum Ramulus and whether there is a substantial difference between Cinnamomi Ramulus and Cinnamomi Cortex.Methods:Differently directional branches of Cinnamomi Ramulus were selected according to the branche cross-sectional diameter(A: 0.3 cm and below;B:0.3~1 cm;C: 1~4 cm).According to the content determination method of cinnamaldehyde in Cinnamomi Cortex and Cinnamomi Ramulus in Chinese Pharmacopoeia(2020 Edition),the content of Cinnamaldehyde in three Cinnamomi Ramulus branches and Cinnamomi Cortex was determined by HPLC.The volatile oil components of Cinnamomi Ramulus A,B,C and Cinnamomi Cortex were extracted by steam distillation method,and were analyzed by gas chromatography-mass spectrometry(GC-MS),the relative contents of these chemical constituents in four volatile oil components were calculated by using peak area normalization method.All the rats except the control group were treated by subcutaneously injecting freunds complete adjuvant(FCA)to establish adjuvant arthritis(AA)model.Five days later,ninety successful model rat were randomly divided into nine groups,including model group,Tripterygium wilfordii polyglycosides group(20 mg/kg),prednisone acetate group(10 mg/kg),three Cinnamomi Ramulus volatile oil groups A,B and C(0.1m L/kg)and three Cinnamomi Cortex volatile oil groups(0.05,0.1and 0.2 m L/kg),another ten rats without modeling were assigned into control group.Rats in control group and model group were received of olive oil(5 m L/kg),and rats in other groups were received different test drugs.all the rats were treated by gastric intubation for twenty-eight consecutive days.The bodyweight,hind paw swelling degree,hind paw swelling index,immune organ index,blood routine,hemorheology index were determined.the serum levels and tissue levels of IL-1β,IL-6,PGE2,TNF-α,VEGF-A,MDA,SOD and NO were analyzed by ELISA.Glass slide method and glass capillary method were used to evaluate the clotting time in each rat on day16.The expression of i NOS and COX-2 in right hind paw were analyzed by WB and the expression of IL-1β,TNF-α and IL-6m RNA in right hind paw were determined by RT-PCR.Results:1.Composition research(1)The content of cinnamaldehyde in Cinnamomi Ramulus A,B,C were1.10%,1.40% and 1.55% respectively,all of which could meet the requirement(cinnamaldehyde,1.0% and above)of Chinese Pharmacopoeia(2020 Edition).The content of Cinnamaldehyde in Cinnamomi Cortex was1.89%,which could meet the requirement(cinnamaldehyde,1.5% and above)of Chinese Pharmacopoeia(2020 Edition).(2)The extraction ratio of the volatile oil in Cinnamomi Ramulus A,B,C and Cinnamomi Cortex were 0.20%,0.30%,0.45% and 1.22% respectively,and forty-three,thirty-six,forty-six and forty-six compounds respectively were isolated and identified,accounting for which were 97.00%,98.49%,98.07%,and 97.65%,and the content of cinnamaldehyde in their volatile oil were 76.61%,77.96%,50.78% and 52.34%.(3)There are 7 components of the total content of Cinnamomi Ramulus C and Cinnamomi Cortex volatile oil with the total relative content of more than 1%,accounting for 83.01% and 82.44%,there was no significant difference in chemical composition between them.Except cinnamaldehyde,there were no other components in the volatile oil of Cinnamomi Cortex and Cinnamomi Ramulus A、B whose total relative content was more than 1%,there were significant differences in chemical constituents among them.(4)There are 4 components of the total content of Cinnamomi Ramulus C and Cinnamomi Cortex volatile oil with the total relative content of more than 1%,accounting for 81.21% and 83.68%,the content of volatile oil in Cinnamomi Ramulus B was slightly higher,and the content of cinnamyl acetate was significantly higher,there are some differences in chemical composition between them,there were significant differences in chemical constituents among them.2.Pharmacodynamic research results(1)Compared with the model group,four volatile oil could obviously increase the bodyweight,Cinnamomi Ramulus C and Cinnamomi Cortex 100μg/m L group were the best.(2)Three volatile oil could obviously decrease the joint swelling degree and secondary joint swelling degree,Cinnamomi Ramulus C group were the best.(3)Four volatile oil could obviously prolong the coagulation time by slide method and glass-tube method,Cinnamomi Ramulus B,C and Cinnamomi Cortex 100 μg/m L group were the best.Four volatile oil could obviously decrease the blood level of WBC,NEUT,LYMP,HPLT and PCT,could obviously improve the hemorheology index.(4)could obviously decrease the serum levels and tissue levels of IL-1β,IL-6,TNF-α,PGE2 and VEGF-A,could obviously decrease serum levels and tissue levels of MDA,NO and increase SOD,Cinnamomi Cortex 0.2 mg/m L group were the best;Among the three groups of Cinnamomi Ramulus volatile oil,Cinnamomi Ramulus C had the most significant effect.(5)Four volatile oil could obviously decrease the expression tissue levels of i NOS and COX-2,could obviously decrease the m RNA expression of IL-1β,TNF-α and IL-6,Cinnamomi Cortex 0.2mg/m L group were the best.Among the three groups of Cinnamomi Ramulus volatile oil,Cinnamomi Ramulus C had the most significant effect.Conclusions:The content of cinnamaldehyde in Cinnamomi Ramulus A,B,C and Cinnamomi Cortex could meet the requirement of Chinese Pharmacopoeia(2020 Edition).There were differences in the chemical components of the volatile oil of Ramulus Cinnamomi A,B and C.but there was no significant difference in the chemical components of between Cinnamomi Ramulus C and Cinnamomi Cortex.The anti-inflammatory effect of thevolatile oil Cinnamomi Cortex was better than that of the volatile oil Cinnamomi Ramulus in three groups.Among the three groups,the volatile oil Cinnamomi Ramulus C had the best anti-inflammatory effect and the volatile oil Cinnamomi Ramulus A had the worst anti-inflammatory effect.The results of hemorheology,anti-inflammatory factors,and the expression of inflammation related proteins and RNA also support the above conclusion,that is,the effect of the volatile oil of Cinnamomi Cortex is more significant than that of Cinnamomi Ramulus in each group,and group Cinnamomi Ramulus C(d = 1 ~ 4 cm)is the best among the three groups.The chemical composition and anti-inflammatory effect of Cinnamomi Ramulus were studied.The results showed that Cinnamomi Ramulus with diameter less than 1cm could be used as Cinnamomi Ramulus,and those with diameter more than 1cm were more similar to that of Cinnamomi Cortex. |
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