CASR/VDR/PTH1R信号通路在含钙肾结石发生机制中的初步研究

李静玲; 柯坤彬; 王振丞; 秦德强; 李颢

doi:10.12259/j.issn.2095-610X.S20220609

CASR/VDR/PTH1R信号通路在含钙肾结石发生机制中的初步研究

doi: 10.12259/j.issn.2095-610X.S20220609

昆明医科大学第一附属医院泌尿外科，云南昆明　650032

基金项目: 云南省高层次卫生技术人才项目（D-2017043）；云南省科技厅科技计划项目（202001AY070001-158）；云南省科技厅基金委员会项目（2019FE001-064）

详细信息

作者简介:
李静玲（1989～），女，湖北蕲春人，住院医师，硕士学位，研究方向：泌尿系结石和肿瘤性疾病

柯坤彬与李静玲对文章有同等贡献

通讯作者:
李颢，E-mail：lihao834@sina.com

中图分类号: R692.4
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出版历程
- 收稿日期: 2022-03-08
- 网络出版日期: 2022-06-07
- 刊出日期: 2022-06-30

A Preliminary Study of CASR/VDR/PTH1R Signaling Pathway in the Pathogenesis of Calcium-containing Kidney Stones

Dept. of Urinary Surgery，The First Affiliated Hospital of Kunming Medical University， Kunming Yunnan 650032，China

摘要

摘要: 目的研究CASR/VDR/PTH1R通路在含钙肾结石发生中的作用及机制。方法取肾结石患者肾脏髓质为实验组，肾脏肿瘤患者正常肾髓质为对照组，分别采用q-PCR、WB、免疫荧光法检测肾实体组织中CASR、VDR、PTH1R的mRNA表达和蛋白水平。结果 CASR、PTH1R基因 mRNA在积水肾髓质及肾肿瘤正常髓质表达水平差异无统计学意义（P > 0.05）。VDR基因的mRNA在积水肾髓质及肾肿瘤正常髓质表达水平差异有统计学意义（P < 0.05）。结论 CASR、VDR、PTH1R在肾结石患者肾脏髓质中表达，且有可能通过CASR/VDR/PTH1R信号通路途径参与含钙肾结石的发生机制。
- 钙敏受体 /
- 维生素D受体 /
- 重组人甲状旁腺激素1受体 /
- 基因表达 /
- 肾结石
Abstract: Objective To study the role and mechanism of CASR, VDR, PTH1R and other genes in the occurrence of calcium-containing kidney stones. Methods The renal medulla of patients with renal calculi was selected as the experimental group, and the normal renal medulla of patients with renal tumors was selected as the control group. The mRNA expression and protein level of CASR, VDR and PTH1R in renal solid tissue were detected by q-PCR, WB and immunofluorescence, respectively. Results There was no significant difference in the expression levels of CASR and PTH1R gene mRNA in the hydronephrotic medulla and the normal medulla of renal tumor （P > 0.05）. The difference of VDR gene mRNA expression in hydronephrotic medulla and normal medulla of renal tumor was statistically significant （P < 0.05）. Conclusion CASR, VDR, and PTH1R are involved in the occurrence of calcium-containing kidney stones, and may participate in the mechanism of calcium-containing kidney stones through the CASR/VDR/PTH1R signaling pathway.
- CASR /
- VDR /
- PTH1R /
- Gene expression /
- Kidney stones

HTML全文

图 1 CASR、VDR、PTH1R的q-PCR箱线图

^*P < 0.05。

Figure 1. Q-PCR box plot of CASR，VDR，PTH1R

下载: 全尺寸图片幻灯片

图 2 CASR、VDR、PTH1R的蛋白质免疫印迹图

Figure 2. Western blot of CASR，VDR，and PTH1R

下载: 全尺寸图片幻灯片

图 3 CASR免疫荧光图（×400）

Figure 3. Immunofluorescence images of CASR（×400）

下载: 全尺寸图片幻灯片

图 4 VDR免疫荧光图（×400）

Figure 4. Immunofluorescence images of VDR（×400）

下载: 全尺寸图片幻灯片

图 5 PTH1R免疫荧光图（×400）

Figure 5. Immunofluorescence images of PTH1R（×400）

下载: 全尺寸图片幻灯片

表 1 引物序列

Table 1. Primer sequence

序列	名称	长度（bp）
TTGCCCTCAACGACCACTTT	GAPDH（H）-F	120
TGGTCCAGGGGTCTTACTCC	GAPDH（H）-R	120
GTCTGGATCTCCTTCATTC	CASR（H）-F	75
GATGGCAATCACCTCTAC	CASR（H）-R	75
TCACTGTATCACCTCTTC	VDR （H）-F	80
GGTCATCTGAATCTTCTTC	VDR （H）-R	80
CAACTACTACTGGATTCTG	PTH1R（H）-F	81
AGGTACTTCTTCTCTGAG	PTH1R（H）-R	81
注：F代表上游引物forward；R代表下游引物reverse；使用LightCycle96进行定量实时聚合酶链式反应（qRTPCR），每个实验中都包括一个阴性对照，一式两份测定所有患者样品和非模板对照，GAPDH为内参。

下载: 导出CSV

表 2 抗体序列表

Table 2. Antibody sequence

抗体	公司	来源	稀释比
CaSR	Santacrus	鼠	1∶1 000
VDR	Proteintech	鼠	1∶5 000
PTH1R	Santa SC-12722	鼠	1∶1 000
GAPDH	abmart P30008M	兔	1∶2 000
一抗	CST 7074		1∶2 000
二抗	CST 7076		1∶2 000

下载: 导出CSV

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