| Objective:Observe allergic rhinitis(Allergic rhinitis,AR)changes of tissue morphology,serum helper T cell 1(Th1)and helper T cell 2(Th2)related cytokines,key molecules of TLR4/p38MAPK/AP-1 signaling pathway and MUC5 AC expression in rat nasal mucosa,In order to explore the mechanism of acupoint injection to correct Th1/Th2 imbalance by affecting TLR4/p38MAPK/AP-1 signaling pathway,and to provide laboratory basis for clinical acupoint injection treatment of AR.Methods:Eight of the 32 SPF SD rats were randomly selected as the normal group,and the rest were sensitized by OVA to replicate the AR rat model.Model rats were randomly divided into model group,non-meridian group and acupoint injection group,with 8rats in each group.Acupoint injection group: “Yintang”(EX-HN3)and bilateral“Yingxiang”(LI20)were injected with dexamethasone and lidocaine equal proportion mixture,0.05 ml per point,once /4 days,4 times in total.Non-transacupuncture and non-acupoint injection group: Three points were selected as injection points at the midpoint of the connection between “Houhai” point and “Huantiao”(GB30)point on the left and right buttocks,5cm below the armpit of the left(or right)forelimb,and the injection drug;dosage;treatment frequency and course were the same as those in the acupoint injection group.The general behavior and nasal symptoms of AR rats were evaluated.The morphological changes of nasal mucosa were observed by HE staining;the serum HA;s Ig E;Ig E;IL-6;IL-8;TNF-α;IL-4 and IFN-γ were detected by ELISA and the nasal mucosa TLR4 and My D88 were detected by immunofluorescence.The expression of p38;p-p38;AP-1 and MUC5 AC proteins in nasal mucosa was detected by WB and the expression of MUC5 AC m RNA was detected by RT-PCR.Results:1.Behavior of rats: After modeling,symptomological scores of model group;non-meridian group and acupoint group were all higher than 5 points,and higher than that of normal group(P<0.05),indicating successful modeling.After treatment,compared with model group,symptomological scores in acupoint injection group and non-meridian and non-acupoint group were significantly decreased(P<0.05),but the difference was not statistically significant(P>0.05).Compared with the non-meridian and non-acupoint group,symptom score of the acupoint injection group was decreased,and the difference was statistically significant(P<0.05).2.HE staining of nasal mucosa tissues: the nasal mucosa structure of rats in the normal group was normal,the connections between columnar epithelial cells were normal,the cilia were arranged neatly,and no mucosal edema or inflammatory cell infiltration was observed.In the model group,nasal mucosa epithelial cells were destroyed and shed;mucosal edema;cilia were disorderly;there were a large number of inflammatory cells infiltrated and submucosal glands and blood vessels proliferated significantly.Compared with the model group,the rats in the non-meridian and non-acupoint group had little change.No obvious shedding of nasal mucosal epithelial cells was observed in the acupoint injection group,the cilia were arranged more neatly,the mucosa was slightly edema,and a few inflammatory cells were infiltrated.Compared with the model group and the non-acupoint injection group,the nasal mucosal epithelial barrier structure was significantly restored.3.ELISA detection results: Compared with normal group,serum HA;s Ig E;Ig E;IL-6;IL-8;TNF-α and IL-4 contents in model group were significantly increased;while serum IFN-γ content was significantly decreased,with statistical significance(P<0.01).Compared with model group,the contents of HA;s Ig E;Ig E;IL-6;IL-8;TNF-α and IL-4 in serum of acupoint injection group were significantly decreased,while the contents of serum IFN-γ were significantly increased,with statistical significance(P<0.01).The contents of HA;s Ig E;Ig E;IL-6;IL-8;TNF-α and IL-4 in the non-meridian and non-acupoint groups were lower than those in the model group,while the contents of IFN-γ in the serum were higher than those in the model group,but the difference was not statistically significant(P>0.05).Compared with the non-meridian and non-acupoint group,the contents of HA;s Ig E;Ig E;IL-6;IL-8;TNF-α and IL-4 in serum of the acupoint injection group were significantly decreased,while the contents of serum IFN-γ were significantly increased,with statistical significance(P<0.01).4.Immunofluorescence detection results: Compared with normal group,positive expressions of TLR4 and My D88 in nasal mucosa of model group were significantly increased,with statistical significance(P<0.01).Compared with model group,the positive expressions of TLR4 and My D88 in nasal mucosa of acupoint injection group were significantly decreased,with statistical significance(P<0.01).The positive expressions of TLR4 and My D88 in nasal mucosa of non-meridian and non-acupoint groups were lower than those of model group,but the difference was not statistically significant(P>0.05).Compared with the non-meridian and non-acupoint group,the positive expressions of TLR4 and My D88 in nasal mucosa of the acupoint injection group were significantly decreased,and the difference was statistically significant(P<0.05,P<0.01).5.Western blot detection results: Positive expressions of p38;p-p38;AP-1 and MUC5 AC were detected in nasal mucosa of rats in all groups.Compared with normal group,positive expressions of p38;p-p38;AP-1 and MUC5 AC in nasal mucosa of model group were significantly increased,with statistical significance(P<0.01).Compared with model group,the positive expressions of p38;p-p38;AP-1 and MUC5 AC in nasal mucosa of acupoint injection group were significantly decreased,with statistical significance(P<0.05,P<0.01).The positive expressions of p38;p-p38;AP-1 and MUC5 AC in nasal mucosa of non-meridian and non-acupoint groups were lower than those of model group,but the difference was not statistically significant(P>0.05).Compared with the non-meridian and non-acupoint group,the positive expressions of p38;p-p38;AP-1 and MUC5 AC in nasal mucosa of the acupoint injection group were significantly decreased,and the difference was statistically significant(P<0.05,P<0.01).6.RT-q PCR detection results: MUC5 AC m RNA was expressed in nasal mucosa of rats in all groups.Compared with normal group,m RNA expression of MUC5 AC in nasal mucosa of model group was significantly increased,with statistical significance(P<0.01).Compared with model group,m RNA expression of MUC5 AC in nasal mucosa of acupoint injection group was significantly decreased,with statistical significance(P<0.01).The m RNA expression of MUC5 AC in nasal mucosa of non-meridian group was lower than that of model group,but the difference was not statistically significant(P>0.05).Compared with the non-meridian and non-acupoint group,the m RNA expression of MUC5 AC in nasal mucosa of the acupoint injection group was significantly decreased,and the difference was statistically significant(P<0.05).Conclusions:1.Acupoint injection can effectively relieve nasal inflammation of AR rats and play a role in the treatment of AR.2.The effect of acupoint injection on AR may be related to down-regulating the expression of HA;Ig E;s Ig E;IL-4;IL-6;IL-8 and TNF-α in serum and up-regulating the expression of IFN-γ,thus correcting the imbalance of Th1/Th2.3.Acupoint injection may play a role in the treatment of AR by inhibiting the abnormal activation of TLR4/p38MAPK/AP-1 signaling pathway,correcting Th1/Th2 imbalance and restoring the structural and functional integrity of nasal mucosal epithelial barrier. |
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