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1.北京中医药大学,北京 100029
2.中日友好医院,北京 100029
3.免疫炎性疾病北京市重点实验室,北京 100029
Published:20 April 2023,
Published Online:10 February 2023,
Received:24 November 2022,
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贺倩,余新波,黄子玮等.化湿润燥方对干燥综合征模型小鼠颌下腺结构和功能的影响[J].中国实验方剂学杂志,2023,29(08):36-44.
HE Qian,YU Xinbo,HUANG Ziwei,et al.Effect of Huashi Runzao Prescription on Structure and Function of Submandibular Gland in Sjögren's Syndrome Model Mice[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(08):36-44.
贺倩,余新波,黄子玮等.化湿润燥方对干燥综合征模型小鼠颌下腺结构和功能的影响[J].中国实验方剂学杂志,2023,29(08):36-44. DOI: 10.13422/j.cnki.syfjx.20231597.
HE Qian,YU Xinbo,HUANG Ziwei,et al.Effect of Huashi Runzao Prescription on Structure and Function of Submandibular Gland in Sjögren's Syndrome Model Mice[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(08):36-44. DOI: 10.13422/j.cnki.syfjx.20231597.
目的
2
探索化湿润燥方对干燥综合征(SS)模型NOD/Ltj小鼠颌下腺组织病理损伤和功能的影响,以及对颌下腺细胞水通道蛋白5(AQP5)表达的调节作用。
方法
2
以NOD/Ltj小鼠构建SS动物模型,选取9周龄雌性NOD/Ltj小鼠随机分为模型组、化湿润燥方组(7.15 g·kg
-1
·d
-1
)、硫酸羟氯喹(HCQ)组(1.30 g·kg
-1
·d
-1
),选取9周龄雌性BALB/c小鼠为正常组,每组6只。给药8周,记录各组小鼠饮水量、唾液流率,观察各组小鼠颌下腺病理染色结果并进行评分,采用免疫组化法(IHC)和蛋白免疫印迹法(Western blot)检测AQP5蛋白的表达水平。
结果
2
与正常组比较,模型组小鼠饮水量明显增加(
P
<
0.05)、唾液流率明显减低(
P
<
0.05)。与模型组比较,化湿润燥方组小鼠饮水量明显降低(
P
<
0.05)、唾液流率明显增加(
P
<
0.05);HCQ组小鼠唾液流率明显增加(
P
<
0.05)。颌下腺组织病理结果显示,与正常组比较,模型组小鼠病理评分、淋巴细胞浸润灶数目、淋巴浸润灶面积百分比均明显增高(
P
<
0.05)。与模型组比较,化湿润燥方组的病理评分、淋巴细胞浸润灶数目均明显降低(
P
<
0.05),化湿润燥方组和HCQ组的淋巴浸润灶面积百分比明显降低(
P
<
0.05)。IHC及Western blot结果显示,与空白组比较,模型组小鼠AQP5蛋白表达水平明显下调(
P
<
0.05)。与模型组和HCQ组比较,化湿润燥方组AQP5蛋白表达水平明显上调(
P
<
0.05)。
结论
2
化湿润燥方可以改善SS模型小鼠颌下腺腺泡细胞的分泌功能和腺体结构损害,其作用机制可能与上调颌下腺细胞AQP5蛋白的表达水平有关。
Objective
2
To investigate the effects of Huashi Runzao prescription (HRP) on the histopathological injury and function of submandibular gland in naive non-obese diabetic (NOD/Ltj) mouse model of Sjögren's syndrome (SS) and its regulatory effect on aquaporin 5 (AQP5) expression in submandibular gland cells.
Method
2
The SS model was induced in NOD/Ltj mice. The NOD/Ltj female mice aged nine weeks were selected and randomly assigned into model group,HRP group (7.15 g·kg
-1
·d
-1
),and hydroxychloroquine (HCQ) group (1.30 g·kg
-1
·d
-1
), and female BALB/c mice in the same age were selected and assigned into the normal group, with six mice in each group. Drug intervention lasted eight weeks. The water consumption and salivary flow rate (SFR) of each group were recorded. The pathological staining results of the submandibular gland of mice in each group were observed and scored. AQP5 expression was determined by immunohistochemistry (IHC) and Western blot.
Result
2
Compared with the normal group, the model group showed increased water consumption (
P
<
0.05) and reduced SFR (
P
<
0.05). Compared with the model group, the HRP group showed decreased water consumption (
P
<
0.05) and increased SFR (
P
<
0.05), and the HCQ group showed increased SFR (
P
<
0.05). In terms of histopathological results of the submandibular gland,compared with the normal group,the model group showed increased pathological score, number of lymphocyte infiltration foci,and percentage of lymphatic infiltration area (
P
<
0.05). Compared with the model group, the HRP group showed reduced pathological scores and number of lymphocyte infiltration foci (
P
<
0.05), and the HRP group and the HCQ group showed reduced percentage of lymphatic infiltration area(
P
<
0.05). The results of IHC and Western blot showed that compared with the normal group,the model group showed down-regulated expression level of AQP5 protein (
P
<
0.05), and compared with the model group and the HCQ group,the HRP group showed up-regulated expression level of AQP5 protein (
P
<
0.05).
Conclusion
2
HRP can improve the secretion function of submandibular gland acinous cells and glandular structure injury in SS model mice, and its mechanism may be related to the up-regulation of AQP5 protein expression level in submandibular gland cells.
化湿润燥方干燥综合征颌下腺NOD/Ltj小鼠水通道蛋白5(AQP5)
Huashi Runzao prescriptionSjögren's syndromesubmandibular glandNOD/Ltj miceaquaporin 5(AQP5)
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