| OBJECTIVE:1.To observe the effect of different quantities of herb-cake-separated moxibustion on immunological function and cellular immunity of immunosuppressed rabbits induced by cyclophosphamide and analyze the differences of the effects of different quantities so as to explore the differences in efficacy and mechanism of different quantities.2.To observe effect of the different time sites of herb-cake-separated moxibustion on immunological function and cellular immunity of immunosuppressed rabbits induced by cyclophosphamide and analysize the latency,optimum and post-response time of efficacy occurrence and development of herb-cake-separated moxibustion in the optimum moxibustion quantities so as to clarify the effect of changes over time,providing thoughts and methods for future clinical research.METHODS:1.50 rabbits were randomly divided into blank group,model group,3 Zhuang,5 Zhuang and 7 Zhuang group of herb-cake-separated moxibustion with 10 rabbits in each group according the method of table of random number.Except the blank group,the other groups were given with intraperitoneal injection of cyclophosphamide(60 mg/kg)to establish immunosuppressive rabbit model.Second days after the end of model,except the blank group and model group,other groups were treated with 3 Zhuang,5 Zhuang and 7 Zhuang of herb-cake-separated moxibustion respectively which herb-cake was made with Liuwei Dihuang decoction in Shenque,Guan Yuan,Zusanli,Pishu and Shenshu,one time every other day,a total of 10 times.The blank group and model group were only tied and fixed to the rabbit platform,without any treatment.The day after the experiment,the experimental animals were weighed and anesthetized,and the celiac venous blood was taken and the spleen was extracted.(1)The contents of serum CD3+,CD4+,CD8+,NK and Bc were measured by ELISA.(2)Spleen quality was weigh and the spleen index was calculated.(3)The contents of tissue IL-2,IL-4 and IFN-γ were measured by ELISA.(4)The morphological changes of spleen in general microscope though the procedure of fixed in 10% formalin,paraffin embedded and HE stained.2.90 rabbits were randomly divided into blank group,model immediate group,model 5 days group,model 10 days group,model 15 days group,immediate after herb-cake-separated moxibustion group,5 days after herb-cake-separated moxibustion group,10 days after herb-cake-separated moxibustion group,15 days after herb-cake-separated moxibustion group with 10 rabbits in each group according the method of table of random number.Except the blank group,the other groups were given with intraperitoneal injection of cyclophosphamide(60 mg/kg)to establish immunosuppressive rabbit model.Second days after the end of model,except the blank group,model immediate group,model 5 days group,model 10 days group and model 15 days group,other groups were treated with herb-cake-separated moxibustion which herb-cake was made with Liuwei Dihuang decoction in Shenque,Guan Yuan,Zusanli,Pishu and Shenshu,one time every other day,a total of 10 times.The blank group and model group were only tied and fixed to the rabbit platform,without any treatment.The second day after the end of the experiment,the experimental animals of the blank group,model immediate group,immediate after herb-cake separated moxibustion group were weighed and anesthetized,and the celiac venous blood was taken and the spleen was extracted.The second day after 5 days,10 days and 15 days of normal feeding of model 5 days group and 5 days after herb-cake separated moxibustion group,model 10 days group and 10 days after herb-cake separated moxibustion group,model 15 days group and 15 days after herb-cake separated moxibustion group respectively,the experimental animals were weighed and anesthetized,and the celiac venous blood was taken and the spleen was extracted.(1)The changes of peripheral blood cell series was detected by blood cell detector.(2)The contents of serum Ig M,Ig G C3 and C4 was measured by immunodetector.The contents of serum CD3+,CD4+,CD8+,NK and Bc were measured by ELISA.(3)Spleen quality was weigh and the spleen index was calculated.(4)The contents of tissue IL-2,IL-4 and IFN-γ were measured by ELISA.(5)The morphological changes of spleen in general microscope though the procedure of fixed in 10% formalin,paraffin embedded and HE stained.RESULTS:1.Compared with the blank group,the weigh D-value after and before experiment was significant decreased;the content of serum CD3+ was significant increased;the content of spleen tissue IFN-γ、IL-4 was significant decreased;the spleen index of model group was significant increased.The results of spleen form metrology shows that compared with the blank group,the average size of white pulp and radius of splenic corpuscle of model group were obvious reduced and lymphatic cells of lymphatic sheath around the arteries of model group were also obvious decreased.The difference was significant statistically.After the treatment of herb-cake-separated moxibustion with different quantities,compared with the model group,the weigh D-value of 3 Zhuang group,5 Zhuang group and 7 Zhuang group after and before experiment was significant increased;the content of serum CD3+ of 3 Zhuang group,5 Zhuang group and 7 Zhuang group was significant decreased;the content of spleen tissue IL-4 of 3 Zhuang group,5 Zhuang group and 7 Zhuang group was significant increased;the spleen index of 3 Zhuang group,5 Zhuang group and 7 Zhuang group was significant increased.The results of spleen form metrology shows that compared with the model group,the average size of white pulp and radius of splenic corpuscle of 3 Zhuang group,5 Zhuang group and 7 Zhuang group were obvious enlarged and lymphatic cells of lymphatic sheath around the arteries of 3 Zhuang group,5 Zhuang group and 7 Zhuang group were also obvious increased.Compared with model group,the content of spleen tissue IFN-γ of 5 Zhuang group and 7 Zhuang group was significant increased.The difference was significant statistically.The effect of herb-cake-separated moxibustion with different quantities on the immune function of immunosuppressed rabbits was different.Compared with the 3 Zhuang group,the weigh D-value of 5 Zhang group and 7 Zhuang group after and before experiment was significant increased;the content of serum CD8+ of 5 Zhuang group was decreased;the content of serum CD3+ of 7 Zhuang group was increased;the content of serum CD8+ of 7 Zhuang group was decreased;the content of tissue IL-4、IFN-γ of 5 Zhuang group and 7 Zhuang group was increased;the average size of white pulp and radius of splenic corpuscle of 5 Zhuang group and 7 Zhuang group were obvious enlarged and lymphatic cells of lymphatic sheath around the arteries of 5 Zhuang group and 7 Zhuang group was increased.The difference was significant statistically.The effect of herb-cake-separated moxibustion with different quantities on the immune function of immunosuppressed rabbits was different.Compared with 5 Zhuang group,the content of serum CD3+ of 7 Zhuang group was significantly increased.The different was no statistically significant between 5 Zhuang group and 7 Zhuang group in the weigh D-value after and before experiment,the content of serum CD8+ and the tissue IFN-γ and IL-4,the average size of white pulp,radius of splenic corpuscle and lymphatic cells of lymphatic sheath around the arteries.The different was no statistically significant among 3 Zhuang group,5 Zhuang group and 7 Zhuang group in spleen index.2.Compared with blank group,the WBC,RBC,HGB,HCT,PLT,NEU percentage,Ig G,Ig M and C3 of model immediate group,model 5 days group,model 10 days group and model 15 days group were significantly decreased(P﹤0.01),while the LYM percentage decreased(P﹤0.01);Compared with blank group,the C4 of model immediate group and model 5 days group was significantly increased(P﹤0.05).Compared with blank group,the content serum CD3+ of model group was decreased and CD8+ was increased;the content of tissue IFN-γ and IL-4 of model group were decreased;the average size of white pulp and radius of splenic corpuscle of model group were induced and lymphatic cells of lymphatic sheath around the arteries of model group was decreased.The difference of WBC,RBC,HGB,HCT,PLT,NEU percentage,LYM percentage,MONO percentage,EOS percentage,BASO percentage,Ig G,Ig M,C3,C4 was no statistically significant among model immediate group,model 5 days group,model 10 days group and model 15 days group.After treatment,compared with model groups at the corresponding time period,the WBC、RBC、HGB、HCT of immediate group,5 days group,10 days group and 15 days group after herb-cake-separated moxibustion were obviously increased and the LYM percentage was decreased(P﹤0.01);the PLT of immediate group,5 days group and 10 days group after herb-cake-separated moxibustion was obviously increased;the NEU percentage of immediate group and 5 days group after herb-cake-separated moxibustion was increased;the content of Ig G 、 Ig M of immediate group,5 days group and 10 days group after herb-cake-separated moxibustion was increased;the content of Ig M of 15 days group after herb-cake-separated moxibustion was increased;the content C3 of immediate group and 5 days group after herb-cake-separated moxibustion was increased;the content C4 of 5 days group after herb-cake-separated moxibustion was increased;the weigh D-value of immediate group,5 days group,10 days group and 15 days group after herb-cake-separated moxibustion after and before experiment was obviously increased and the spleen index was decreased.Compared with model group,the content of serum CD3+ of immediate group,5 days group,10 days group and 15 days group after herb-cake-separated moxibustion was obviously increased and CD8+ of immediate group,5 days group,10 days group and 15 days group after herb-cake-separated moxibustion was obviously decreased;the content of tissue IFN-γ、IL-4 of immediate group,5 days group,10 days group and 15 days group after herb-cake-separated moxibustion was obviously increased;the average size of white pulp and radius of splenic corpuscle of immediate group,5 days group,10 days group and 15 days group after herb-cake-separated moxibustion were enlarged and lymphatic cells of lymphatic sheath around the arteries was increased.Compared with the immediate group and 5 days group after herb-cake-separated moxibustion,WBC、RBC、HGB、HCT of 10 days after herb-cake-separated moxibustion was obviously increased and LYM percentage was decreased;the content of Ig G,Ig M of 10 days after herb-cake-separated moxibustion was obviously increased;the weigh D-value of 10 days after herb-cake-separated moxibustion after and before treatment was obviously increased;the spleen index of 10 days after herb-cake-separated moxibustion was obviously decreased;the content of serum CD3+ of 10 days after herb-cake-separated moxibustion was obviously increased and CD8+ of 10 days after herb-cake-separated moxibustion was decreased;the content of tissue IFN-γ and IL-4 of 10 days after herb-cake-separated moxibustion was obviously increased;the average size of white pulp and radius of splenic corpuscle of 10 days after herb-cake-separated moxibustion were enlarged.Compared with the immediate group,the WBC of 5 days and 10 days after herb-cake-separated moxibustion was obviously increased and the NEU percentage was obviously decreased;the content of C3 of 5 days after herb-cake-separated moxibustion was obviously increased,while the content of C3 of 10 days and 15 days after herb-cake-separated moxibustion decreased;the lymphatic cells of lymphatic sheath around the arteries of 15 days group after herb-cake-separated moxibustion was increased;the weigh D-value of 15 days group after herb-cake-separated moxibustion after and before treatment was obviously increased.Compared with 5 days group after herb-cake-separated moxibustion,the content of C3 of 10 days group and 15 days after herb-cake-separated moxibustion was decreased;the radius of splenic corpuscle of 15 days group after herb-cake-separated moxibustion was induced.Compared with 10 days group,the WBC、RBC、HGB、HCT、PLT of 15 days after herb-cake-separated moxibustion were decreased,while the LYM percentage of 15 days after herb-cake-separated moxibustion was increased;the Ig G and Ig M of 15 days after herb-cake-separated moxibustion were obviously decreased;the spleen index of 15 days after herb-cake-separated moxibustion was obviously increased;the content of serum CD3+ was decreased,while the content of serum CD8+ increased;the content of tissue IFN-γ and IL-4 of 15 days after herb-cake-separated moxibustion were decreased;the average size of white pulp and radius of splenic corpuscle of 15 days after herb-cake-separated moxibustion were induced.CONCLUSIONS:1.Herb-cake-separated moxibustion with different quantities on on immune cells,immune molecules and immune organs of immunosuppressed rabbits induced by cyclophosphamide has obviously regulatory effects.2.The effect of herb-cake-separated moxibustion with different quantities on immune cells,immune molecules and immune organs of immunosuppressed rabbits induced by cyclophosphamide was different,and there is a significant dose-effect relationship.5 Zhuang and 7 Zhuang moxibustion amount on the immune function of immunosuppressed rabbits was more obvious than that of 3 Zhuang.The best moxibustion amount in this project is 5 strong.3.The effect of herb-cake-separated moxibustion with 5 Zhuang on immune cells,immune molecules and immune organs of immunosuppressed rabbits induced by cyclophosphamide has time-effect relationship.From the analysis of the change trend of a number of indicators,taking effect immediately after herb-cake-separated moxibustion,reaching the optimum effect 10 days after herb-cake-separated moxibustion,efficacy began to decay subsequently.4.From the overall effect of herb-cake-separated moxibustion on on immune cells,immune molecules and immune organs of immunosuppressed rabbits induced by cyclophosphamide has obviously dose-effect and time-effect relationship.Herb-cake-separated moxibustion with 5 Zhuang is the optimum moxibustion quantitiy and the optimum efficacy occurance on the tenth after herb-cake-separated moxibustion. |
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