| 1 Objective In order to discuss the effect of moxa smoking,and olfactory tract mediate the moxa smoking effect,and based on the clinical application of moxibustion,to observe the changes of injury of knee osteoarthritis model rats under different density of moxa smoke,whether the olfactory tract had role in regulating the functions of the body,and to discuss the effect of different density of moxa smoking on the olfactory bult,to offer experimental support for deeply discuss the mechanism of moxa smoke and the moxibustion.2 Methods Apply phormocological,neuroelectrophysiological and molecular biology methods to observe the ultra-structure changes of cartilage,TNF-a,IL-1b,BAX and Bal-2 m RNA expressions in tissue fluids of different density of moxa smoke on KOA model rats,to discuss the effectivness of moxa smoke.To observe the ultra-structure of olfactory bulb and the expression of c-Fos in OB of KOA model rats,to discuss the safety of moxa smoke.To observe the changes of 5-HT,DA,NE and Ach in brain after olfactory bulb ectomy,to discuss the mechanism of olfactory tract mediated moxa smoking effect.To compare the different active potentials in olfactory bulb with moxa oil,moxa smoke and smoke stimulation to discuss the possible mechanism of olfactory tract.3 Results 1)87 mass spector spike were detected by HS-GC-MS,and 54 chemical components were identified,which is 88.505% to totally identified materials.And in the moxa smoke,mainly volatile componts were alkanes,alkenes,aldehydes,acids,esters,phenols,terpenoids and heterocyclic chemicals.2)To simultate the clinical circumstance,and detect the density of moxa smoke with “open the windows” methods with environment detection,to make 0.11mg/m3,0.23mg/m3 and 0.45mg/m3 of moxa somke to be the lower,middle and higher density to make further experiments.3)Under transmission electron microscopy at a magnification of 2000 x,articular cartilage cells in the normal group were ovoid,cell membranes were complete and microvilli were noted.Nuclei were normal with smooth nuclear membranes and homogeneous chromatin.Mitochondria,Golgi complex and rough endoplasmic reticulum were intact.In the model control group,cartilage cells were irregular in shape.Lipid droplets noted.Nuclear disruption was present.,Organelles such as mitochondria and rough endoplasmic reticulum were missing.Free cytosol ribosomes were significantly decreased.In the low particle density exposure group,the morphology of cartilage cells was irregular.Cellular nuclei showed pyknosis.Mitochondria were decreased.Rough endoplasmic reticulum evidenced signs of fracturing and disintegration.In the middle density group,abnormal cartilage cells were also observed.Chromatin was found to be migrating to the periphery of the nuclei,Mitochondria were decreased and the rough endoplasmic reticulum showed signs of expansion and fracturing.In the high density group,cartilage cells were swollen.Microvilli were shortened and fractured.Mitochondria had signs of vacuolization.The rough endoplasmic reticulum showed expansion.4)Compare to the normal control group,the expression of TNF-a,IL-1b and BAX m RNA were significantly increased in model control group(P<0.01),and the expression of Bcl-2 m RNA decreased greatly(P<0.01).Compare to the model control group,after intervened in different density of moxa smoke,the expression of TNF-a,IL-1b and BAX m RNA in the joint fluid were decreased,while Bcl-2 increased.Under the middle and higher density of moxa smoke circumstance,the change of expression of TNF-a,IL-1b,BAX and Bcl-2 m RNA were significance.The ratio of Bcl-2 m RNA/BAX m RNA in model control group were also decreased greatly(P<0.01),and in the different density of moxa smoke,the ratio increased gradually,and to compared to in the model control group,the ratio in middle and higher density of moxa smoke group were increased significantly.5)To observe the olfactory bulb with transmission electron microscopy at a magnification of 6000 x and 20000 x,in the normal control group,the complete cell,membrane,and nuclei were obviously,the membrane of nuclei was clearly,chromatin and ribosome were homogeneous,and lysosome,mitochondria,and plenty of rough endoplasmic reticulum were intact.In the model control group of olfactory bulb,the membrane of cell were completely,inflation of nuclear membrane and edema in cytoplasm,the mitochondria expensed and degeneration,and rough reticulum were missing.In the lower density of moxa smoke,completely cell and membrand were observed,inflated nuclear membrand and glycogen redused,and little mitochondria expensed and degeneration,rough reticulum were breakage.In the middle density group,the nuclear membrand were complete and clearly,a lot of rough reticulum without expension but a little were ruptured,a little mitochondrial crest fractured.In the higher density group,a clearly cell and membrane were seen,the nuclear were completely with homogeneous chromatin,clearly ribosome,lysosome,and mitochondrial in cytoplasm,and plenty of rough reticulum with little ruptured.6)A lot of c-Fos marked proteins in every granular layer of olfactory bulb,and no significance variation in each group.In different ambient density of moxa smoke,such conditions were observed,but to compare to in the model control group,varation were not significance.7)To compare with in normal control group,5-HT,DA,NE and Ach in brain inmodel control group were decreased,but not obviously(P>0.05).To compare to in the model control group,the content of 5-HT,DA and Ach in higher density of moxa smoke were increased significantly(P<0.01),and the content of NE increased without significance difference.After olfactory bulbectomy,the content of 5-HT,DA,NE and Ach in the brain increased.Compare to in the moxa smoking group,the content of DA and Ach decreased obviously(P<0.01),but no significance varations of the content of 5-HT and NE(P>0.05).8)At the time of smell moxa oil,the cone cells in olfactory bulb discharged 22.0±1.3 count/bin,and single neuron firing were 0.36±0.01 spikes/sec.At the time of higher density of moxa smoke intervened,the cone cells in olfactory bulb discharged 17.67±0.82 count/bin,and single neuron firing were 0.28±0.01 spikes/sec,the number were 11.67±1.37 count/bin and 0.22±0.01spikes/sec respectively in the intervene of smoking.There were significantly varations in each group(P<0.05).4 Conclusion 4.1 Different density of moxa smoke could change the ultra-structure of cartilage in KOA model rats,and improve the inflammatory conditions and protect the cell above the apoptosis.4.2 Moxa smoke could improve the active of OB,and odor moleculos were the main gredients.4.3 Moxa smoke could help to release aminoamine neurotransmitters and acetyl choline in the brain,which could have role in regulating the brain functions,and the olfactory tract,that based on the olfactory bulb,take part in the regulatin effect of moxa smoke. |
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