润燥解毒通络法对干燥综合征小鼠模型血清及颌下腺M3R表达的影响

发布时间：2018-04-02 14:26

本文选题：润燥解毒通络法　切入点：干燥综合征　出处：《浙江中医药大学》2017年硕士论文

【摘要】：目的研究润燥解毒通络法中药对干燥综合征(sjogren syndrome,SS)小鼠模型血清及颌下腺毒蕈碱样乙酰胆碱3型(muscarinic acethyleholine receptor 3,M3R)表达的影响,以探讨润燥解毒通络法中药(解毒通络生津汤)治疗SS的作用机理。方法选用BALB/c小鼠47只,随机分为正常对照组(正常组,n=10)和造模组(n=37)。取BALB/c小鼠颌下腺制备抗原进行造模。造模组采用背部皮内5个部位多点注射抗原的方法进行免疫,建立SS模型。造模成功后随机分为模型组(n=10)、中药组(n=10)和HCQ组(n=10)。分别灌胃给药:中药组按11.4g/kg/d剂量灌胃中药解毒通络生津汤,模型组灌胃等量生理盐水,HCQ组小鼠按0.06g/kg/d剂量灌胃HCQ混悬液(HCQ与生理盐水混合液),均每日1次,正常组正常饲养。同时测定小鼠唾液量、饮水量。连续用药30天后处死,取各组小鼠颌下腺、脾及血清。采用苏木素-伊红(HE)染色评价颌下腺淋巴细胞浸润情况,采用ELISA法测定标本血清M3R水平,免疫组化法检测颌下腺M3R表达水平。结果1、造模成功后治疗前,与正常组比较,模型组、中药组和HCQ组小鼠唾液量减少,差异均有统计学意义(P0.05);模型组、中药组、HCQ组之间比较,小鼠唾液量比较差异无统计学意义(P0.05)。治疗第30天,与正常组比较,模型组唾液量减少,差异有统计学意义(P0.05);与模型组比较,中药组、HCQ组小鼠唾液量增加,差异均有统计学意义(P0.05);中药组和HCQ组之间比较差异无统计学意义(P0.05)。与本组治疗前比较,治疗后中药组、HCQ组小鼠唾液量增加,差异均有统计学意义(P0.05)。2、与正常组比较,模型组小鼠血清M3R表达增加,差异有统计学意义(P0.05);与模型组比较,中药组、HCQ组M3R表达减少,差异有统计学意义(P0.05)。中药组和HCQ组之间比较,差异无统计学意义(P0.05)。3、与正常组比较,模型组小鼠颌下腺M3R表达增加,差异有统计学意义(P0.01);与模型组比较,中药组、HCQ组小鼠颌下腺M3R表达减少,差异有统计学意义(P0.05)。中药组和HCQ组之间比较,差异无统计学意义(P0.05)。4、与正常组比较,模型组小鼠颌下腺淋巴细胞浸润明显,差异有统计学意义(P0.01);与模型组比较,中药组和HCQ组小鼠颌下腺淋巴细胞浸润减轻,差异有统计学意义(P0.05);中药组和HCQ组之间比较,差异无统计学意义(P0.05)。5、各组之间脾指数、颌下腺指数比较,差异均无统计学意义(P0.05)。结论SS小鼠血清及颌下腺M3R表达水平升高。润燥解毒通络法中药可能通过下调M3R表达,从而抑制颌下腺淋巴细胞浸润,改善腺体分泌能力,阻止SS进展。
[Abstract]:Objective to study the effect of moistening and detoxifying and removing collaterals on the expression of muscarinic acethyleholine receptor 3 M 3R in serum and submandibular gland of Sjogren syndrome (Sjogren syndromes) mouse model.To explore the mechanism of moistening dryness and detoxification and dredging collaterals (detoxifying and releasing collaterals Shengjin decoction) in treating SS.Methods 47 BALB/c mice were randomly divided into normal control group (normal group) and model group.The submandibular glands of BALB/c mice were taken to prepare antigens for modeling.The model group was immunized by multiple injection of antigens in 5 parts of the back skin, and SS model was established.The models were randomly divided into two groups: model group (n = 10), Chinese medicine group (n = 10) and HCQ group (n = 10).The rats in the model group were treated with the same amount of normal saline. The mice in the model group were fed with the mixture of HCQ and normal saline at the same dose of 0.06g/kg/d, all of them were fed once a day, and the normal control group was fed normally.At the same time, the amount of saliva and drinking water were determined.After 30 days of continuous administration, the mice were killed and the submandibular gland, spleen and serum were taken.The submandibular gland lymphocytic infiltration was evaluated by hematoxylin and eosin (HEH) staining, the serum M3R level was measured by ELISA method, and the expression of M3R in submandibular gland was detected by immunohistochemical method.Results 1. Before treatment, the saliva quantity of model group, traditional Chinese medicine group and HCQ group decreased significantly compared with normal group, and there was no significant difference between model group and traditional Chinese medicine group (P 0.05).On the 30th day, compared with the normal group, the saliva quantity in the model group decreased, the difference was statistically significant (P 0.05), compared with the model group, the saliva quantity in the HCQ group increased.The difference was statistically significant (P 0.05), but there was no significant difference between traditional Chinese medicine group and HCQ group (P 0.05).Compared with the control group, the saliva content of the HCQ group increased after treatment, and the difference was statistically significant (P 0.05). Compared with the normal group, the serum M3R expression in the model group was higher than that in the normal group (P 0.05), and that in the model group was significantly higher than that in the model group.The expression of M3R in HCQ group was significantly lower than that in HCQ group (P 0.05).Compared with the normal group, the expression of M3R in the submandibular gland of the model group was increased, the difference was statistically significant (P 0.01), and the expression of M3R in the submandibular gland of the Chinese medicine group was lower than that of the model group, while the expression of M3R in the submandibular gland of the model group was significantly lower than that in the control group.The difference was statistically significant (P 0.05).There was no significant difference between the traditional Chinese medicine group and the HCQ group, and there was no significant difference between the two groups. Compared with the normal group, the submandibular gland lymphocytic infiltration in the model group was obvious, the difference was statistically significant (P 0.01), and that in the model group was higher than that in the model group.The lymphocyte infiltration of submandibular gland in Chinese medicine group and HCQ group was alleviated, the difference was statistically significant (P 0.05), but there was no significant difference between traditional Chinese medicine group and HCQ group. There was no significant difference in spleen index and submandibular gland index between each group.Conclusion the expression of M 3 R in serum and submandibular gland of SS mice was increased.The method of moistening dryness detoxification and dredging collaterals may inhibit the lymphocyte infiltration of submandibular gland improve the secretory ability of glands and prevent the progression of SS by down-regulating the expression of M3R.
【学位授予单位】：浙江中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5;R-332

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