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1.北京中医药大学 东方医院,北京 100078
2.北京中医药大学 东直门医院 通州院区,北京 101121
李方凯,硕士,主治医师,从事中医药防治风湿免疫疾病研究,E-mail:lifangkai1324@163.com
* 沈潜,博士,副主任医师,从事中医药防治风湿免疫疾病研究,E-mail:rbd144@163.com
收稿日期:2021-08-06,
网络出版日期:2021-10-29,
纸质出版日期:2021-12-20
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李方凯,朱跃兰,侯秀娟等.活血解毒润燥方对干燥综合征小鼠颌下腺细胞凋亡抑制因子Survivin表达的影响[J].中国实验方剂学杂志,2021,27(24):84-91.
LI Fang-kai,ZHU Yue-lan,HOU Xiu-juan,et al.Effect of Huoxue Jiedu Runzao Prescription on Expression of Apoptosis Inhibitor Survivin in Submandibular Gland Cells of Sjögren's Syndrome Mice[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(24):84-91.
李方凯,朱跃兰,侯秀娟等.活血解毒润燥方对干燥综合征小鼠颌下腺细胞凋亡抑制因子Survivin表达的影响[J].中国实验方剂学杂志,2021,27(24):84-91. DOI: 10.13422/j.cnki.syfjx.20212438.
LI Fang-kai,ZHU Yue-lan,HOU Xiu-juan,et al.Effect of Huoxue Jiedu Runzao Prescription on Expression of Apoptosis Inhibitor Survivin in Submandibular Gland Cells of Sjögren's Syndrome Mice[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(24):84-91. DOI: 10.13422/j.cnki.syfjx.20212438.
目的
2
以干燥综合征(SS)模型小鼠颌下腺细胞凋亡抑制因子(Survivin)的表达为靶点,探讨活血解毒润燥方对SS模型小鼠颌下腺组织形态、细胞凋亡、腺体功能的影响及其干预机制。
方法
2
实验选取雌性BALB/c57小鼠为正常组,自发性雌性非肥胖型糖尿病(NOD/Ltj)小鼠为SS模型并随机分为模型组、白芍总苷胶囊组(0.234 g·kg
-1
)和活血解毒润燥方低、中、高剂量(15.6,31.2,62.4 g·kg
-1
)组,每组15只。药物干预8周后,观察各组小鼠颌下腺形态、功能和凋亡抑制因子Survivin表达的改变,分别采用免疫组化法(IHC),蛋白免疫印迹法(Western blot),逆转录聚合酶链式反应(RT-PCR)检测细胞凋亡抑制因子Survivin蛋白及mRNA的表达。
结果
2
与正常组比较,模型组小鼠唾液流量及颌下腺指数均显著降低(
P<
0.01),颌下腺组织病理学评分显著升高(
P<
0.01),Western blot结果显示,Survivin蛋白表达明显降低(
P<
0.05),IHC结果显示,Survivin表达显著降低(
P<
0.01),RT-PCR结果显示,Survivin mRNA表达显著降低(
P<
0.01);与模型组比较,活血解毒润燥方各剂量组和白芍总苷胶囊组小鼠唾液流量及颌下腺指数均明显增高(
P<
0.05),颌下腺组织病理学评分明显降低(
P<
0.05),IHC结果显示,Survivin表达显著升高(
P<
0.01),Western blot和RT-PCR结果显示,活血解毒润燥方高剂量组与白芍总苷胶囊组Survivin蛋白及mRNA表达均明显升高(
P<
0.05)。
结论
2
活血解毒润燥方可以改善SS模型小鼠颌下腺腺泡的分泌功能,提高其颌下腺指数,增加SS模型鼠的唾液分泌量;其作用机制与上调SS模型鼠颌下腺细胞凋亡抑制因子Survivin的表达水平有关。
Objective
2
To investigate the effect of Huoxue Jiedu Runzao prescription on the morphology, apoptosis, and function of submandibular gland in the mouse model of Sjögren's syndrome (SS) and its functioning mechanism, we analyzed the expression of the apoptosis inhibitor Survivin in the submandibular gland cells of SS mice.
Method
2
Female BALB/c57 mice were selected as the normal group. The naive non-obese diabetic (NOD/Ltj) female mice were selected as the SS model, which were randomly assigned into the model group, Paeoniae Radix Alba total glucosides capsule (0.234 g·kg
-1
) group, and low-, medium-, and high-dose (15.6, 31.2, 62.4 g·kg
-1
, respectively) Huoxue Jiedu Runzao prescription groups. Each group had 15 mice. The morphological and functional changes of submandibular gland and the Survivin expression were observed and measured after 8 weeks of drug intervention. Survivin expression was determined by immunohistochemistry (IHC), Western blot, and reverse transcription-polymerase chain reaction (RT-PCR).
Result
2
Compared with normal group, salivary flow and submandibular gland index in model group were significantly decreased (
P
<
0.01), and histopathological score of submandibular gland was significantly increased (
P
<
0.01). Western blot showed that Survivin protein expression was significantly decreased (
P
<
0.05). IHC showed that, Survivin mRNA expression was significantly decreased (
P
<
0.01), and RT-PCR results showed that Survivin mRNA expression was significantly decreased (
P
<
0.01). Compared with model group, salivary flow and submandibular gland index of mice in Huoxue Jiudu Runzao prescription groups and Paeoniae Radix Alba total glucosides capsules groups were significantly increased (
P
<
0.05), histopathological score of submandibular gland was significantly decreased (
P
<
0.05), IHC results showed that Survivin expression was significantly increased (
P
<
0.01). Western blot and RT-PCR results showed that Survivin protein and mRNA expression of Huoxue Jiudu Runzao prescription high-dose group and Paeoniae Radix Alba total glucosides capsule group were significantly increased (
P
<
0.05).
Conclusion
2
Huoxue Jiedu Runzao prescription can improve the secretion function of submandibular acinus, increase the submandibular gland index, and saliva secretion of SS mice by up-regulating survivin in submandibular gland cells of SS mice.
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