去卵巢和雌激素預(yù)防性給藥對(duì)大鼠甲狀旁腺激素基因的表達(dá)調(diào)控研究
發(fā)布時(shí)間：2018-07-26 11:38
【摘要】：甲狀旁腺是機(jī)體內(nèi)分泌腺之一,其主要功能是分泌甲狀旁腺激素(parathyroid hormone,PTH)。雌激素(estrogen)是由卵巢分泌的一種類固醇激素,雌激素的降低會(huì)打破鈣磷平衡而導(dǎo)致骨質(zhì)疏松,而PTH是人體骨轉(zhuǎn)換率的主要決定因素,表明雌激素與PTH之間可能存在某種聯(lián)系。然而,迄今為止尚缺乏雌激素調(diào)控PTH分泌的直接證據(jù),雌激素調(diào)控PTH分泌的機(jī)理尚不清楚。本研究采用雙側(cè)卵巢切除術(shù)建立絕經(jīng)后骨質(zhì)疏松(postmenopausal osteoporosis,PMOP)大鼠模型,利用石蠟組織切片和HE染色對(duì)大鼠的股骨、脛骨、子宮、十二指腸、腎皮質(zhì)和腎髓質(zhì)進(jìn)行組織形態(tài)學(xué)觀察,利用酶聯(lián)免疫吸附測(cè)定法(enzyme-linked immunosorbent assay,ELISA)檢測(cè)血清中雌激素的變化,用實(shí)時(shí)定量PCR(real-time quantitative polymerase chain reaction,Real-time qPCR)技術(shù)檢測(cè)SD大鼠甲狀旁腺中PTH、雌激素受體(estrogen receptor,ER)表達(dá)變化。進(jìn)一步對(duì)調(diào)控PTH表達(dá)的潛在通路中的關(guān)鍵基因,如鈣敏感受體(calcium sensing receptor,CaSR)、維生素D受體(vitamin D receptor,VDR)、成纖維細(xì)胞生長(zhǎng)因子-23(fibroblast growth factor-23,FGF-23)、FGF-23的受體1(fibroblast growth factor receptor 1,FGFR1)、FGF-23的輔助受體Klotho和早期生長(zhǎng)反應(yīng)基因1(early growth response gene 1,Egr1)的表達(dá)量進(jìn)行分析,探討去卵巢和預(yù)防性給藥對(duì)PTH的調(diào)控機(jī)制。臟器和骨組織形態(tài)學(xué)觀察結(jié)果表明,去卵巢(ovariectomized,OVX)組大鼠的股骨和脛骨均出現(xiàn)骨小梁斷裂、間距變大、結(jié)構(gòu)紊亂等骨質(zhì)疏松癥狀,而去卵巢后預(yù)防性注射β-雌二醇(β-estradiol-treated OVX,OVX/E2)組并未出現(xiàn)明顯的病理癥狀。相較于假手術(shù)(Sham-operated,Sham)組,OVX組和OVX/E2組的十二指腸、腎皮質(zhì)和腎髓質(zhì)區(qū)都未出現(xiàn)明顯病變。相較于Sham組,OVX組大鼠子宮內(nèi)膜固有層中的子宮腺數(shù)目增多、腺腔增大、子宮黏膜上皮明顯增厚。OVX/E2組的子宮腺腺腔雖略有增大,但腺體形態(tài)較為規(guī)則,其子宮內(nèi)膜也未發(fā)生明顯的增厚現(xiàn)象。說明注射小劑量的β-雌二醇可以預(yù)防骨質(zhì)疏松癥的發(fā)生,且不會(huì)有子宮癌變的現(xiàn)象發(fā)生。ELISA檢測(cè)結(jié)果表明三種不同處理組間大鼠的血清雌激素沒有顯著性的差異。Real-time qPCR檢測(cè)結(jié)果表明,去卵巢后大鼠PTH的表達(dá)量顯著下調(diào),而預(yù)防性注射β-雌二醇后PTH的表達(dá)量顯著上調(diào)。ER-α存在于甲狀旁腺中,且去卵巢后ER-α的表達(dá)量上調(diào),而預(yù)防性注射β-雌二醇后ER-α的表達(dá)量顯著下調(diào),說明甲狀旁腺中ER-α能夠響應(yīng)雌激素的微弱變化,參與PTH的表達(dá)調(diào)控。去卵巢后大鼠甲狀旁腺中VDR的表達(dá)量上調(diào),而預(yù)防性注射β-雌二醇后大鼠甲狀旁腺中VDR的表達(dá)量下調(diào),表明VDR可能是抑制PTH的表達(dá)。去卵巢后血清中Ca~(2+)略有升高,甲狀旁腺中CaSR的表達(dá)量顯著下調(diào),預(yù)防性注射β-雌二醇后Ca~(2+)略有升高,甲狀旁腺中CaSR的表達(dá)量顯著上調(diào),表明Ca~(2+)/CaSR調(diào)控系統(tǒng)可能參與PTH的表達(dá)調(diào)控。去卵巢后大鼠甲狀旁腺中FGF-23、FGFR1、Klotho及Egr1表達(dá)量顯著下調(diào),而預(yù)防性注射β-雌二醇后FGF-23、Klotho和Egr1的表達(dá)量下調(diào),FGFR1的表達(dá)量顯著上調(diào),說明去卵巢后甲狀旁腺中FGF-23/Klotho系統(tǒng)對(duì)PTH表達(dá)的抑制作用降低了,但是預(yù)防性注射β-雌二醇后這個(gè)受體系統(tǒng)的功能更低。這些結(jié)果表明,去卵巢和預(yù)防性給藥可能會(huì)影響甲狀旁腺中ER-α、VDR信號(hào)調(diào)控系統(tǒng)、Ca~(2+)/CaSR調(diào)控系統(tǒng)以及FGF-23/Klotho受體調(diào)控系統(tǒng),進(jìn)而調(diào)控PTH的表達(dá)。
[Abstract]:Parathyroid gland is one of the endocrine glands of the body, its main function is the secretion of parathyroid hormone (PTH). Estrogen (estrogen) is a steroid hormone secreted by the ovary. The decrease of estrogen will break the balance of calcium and phosphorus and cause osteoporosis, and PTH is the main determinant of bone turnover rate in human body, indicating that estrogen and P are the main factor. There may be some connection between TH. However, so far, there is no direct evidence that estrogen regulates the secretion of PTH. The mechanism of estrogen regulation of PTH secretion is not clear. This study uses bilateral oophorectomy to establish postmenopausal osteoporosis (postmenopausal osteoporosis, PMOP) rats model, using paraffin tissue section and HE staining to large The histomorphology of the femur, tibia, uterus, duodenum, kidney cortex and renal medulla were observed and the changes of estrogen in serum were detected by enzyme-linked immunosorbent assay (ELISA), and the real-time quantitative PCR (real-time quantitative polymerase chain reaction, Real-time qPCR) was used to detect the size of the rat. The changes in the expression of PTH and estrogen receptor (ER) in the parathyroid gland of the rat are the key genes in the potential pathways regulating the expression of PTH, such as the calcium sensitive receptor (calcium sensing receptor, CaSR), the vitamin D receptor (vitamin D), and fibroblast growth factor. The expression of the receptor 1 (fibroblast growth factor receptor 1, FGFR1), FGF-23 auxiliary receptor Klotho and the early growth response gene 1 (early growth response gene 1, Egr1) was analyzed to explore the regulation mechanism of the ovariectomized and preventive administration. In the femur and tibia of the rats, the fracture of the trabecular bone, the spacing and the disorder of the structure were observed. The prophylactic injection of beta estradiol (beta -estradiol-treated OVX, OVX/E2) after the ovariectomy had no obvious pathological symptoms. Compared to the sham operation (Sham-operated, Sham) group, the duodenum, the renal cortex and the renal cortex of group OVX and OVX/E2 group. There were no obvious lesions in the medullary region. Compared to group Sham, the number of uterine glands in the lamina propria of group OVX increased, the gland cavity increased, and the epithelium of the uterus was thickened in.OVX/E2 group, although the adeno gland of the uterus increased slightly, but the shape of the gland was more regular, and the endometrium had no obvious thickening. The amount of beta estradiol can prevent the occurrence of osteoporosis, and there is no.ELISA detection of the phenomenon of uterine carcinogenesis. The results of serum estrogen in the three different treatment groups showed no significant difference in serum estrogen. The results of.Real-time qPCR detection showed that the amount of PTH in the ovariectomized rats was significantly down, while the prophylactic injection of beta female was a prophylactic injection. After alcohol, the expression of PTH was significantly up-regulated in parathyroid glands, and the expression of ER- alpha was up-regulated after ovariectomized, while the expression of ER- alpha was significantly downregulated after the prophylactic injection of beta estradiol. It indicated that ER- alpha in parathyroid glands could respond to the weak changes in estrogen and participate in the expression and regulation of PTH. The expression of VDR in parathyroid glands of ovariectomized rats The amount of VDR was down regulated in the parathyroid parathyroid after prophylactic injection of beta estradiol, indicating that VDR may be the expression of inhibition of PTH. The serum Ca~ (2+) increased slightly after ovariectomized, and the expression of CaSR in parathyroid glands decreased significantly. The Ca~ (2+) increased slightly after the prophylactic injection of beta estradiol, and the expression of CaSR in parathyroid glands was significantly higher. The Ca~ (2+) /CaSR regulatory system may be involved in the regulation of PTH expression. The expression of FGF-23, FGFR1, Klotho and Egr1 in the parathyroid glands of the ovariectomized rats is significantly down, while the prophylactic injection of beta estradiol is down to FGF-23, Klotho and Egr1, and the expression of FGFR1 is up regulated, indicating the line in the parathyroid gland after ovariectomized. The inhibition of PTH expression is reduced, but the function of the receptor system after a prophylactic injection of beta estradiol is lower. These results suggest that ovariectomy and preventive administration may affect the ER- alpha, the VDR signal regulation system, the Ca~ (2+) /CaSR regulatory system and the FGF-23/Klotho receptor regulation system, and then regulate the table of PTH. Da.
【學(xué)位授予單位】：陜西理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R580
本文編號(hào)：2145890
[Abstract]:Parathyroid gland is one of the endocrine glands of the body, its main function is the secretion of parathyroid hormone (PTH). Estrogen (estrogen) is a steroid hormone secreted by the ovary. The decrease of estrogen will break the balance of calcium and phosphorus and cause osteoporosis, and PTH is the main determinant of bone turnover rate in human body, indicating that estrogen and P are the main factor. There may be some connection between TH. However, so far, there is no direct evidence that estrogen regulates the secretion of PTH. The mechanism of estrogen regulation of PTH secretion is not clear. This study uses bilateral oophorectomy to establish postmenopausal osteoporosis (postmenopausal osteoporosis, PMOP) rats model, using paraffin tissue section and HE staining to large The histomorphology of the femur, tibia, uterus, duodenum, kidney cortex and renal medulla were observed and the changes of estrogen in serum were detected by enzyme-linked immunosorbent assay (ELISA), and the real-time quantitative PCR (real-time quantitative polymerase chain reaction, Real-time qPCR) was used to detect the size of the rat. The changes in the expression of PTH and estrogen receptor (ER) in the parathyroid gland of the rat are the key genes in the potential pathways regulating the expression of PTH, such as the calcium sensitive receptor (calcium sensing receptor, CaSR), the vitamin D receptor (vitamin D), and fibroblast growth factor. The expression of the receptor 1 (fibroblast growth factor receptor 1, FGFR1), FGF-23 auxiliary receptor Klotho and the early growth response gene 1 (early growth response gene 1, Egr1) was analyzed to explore the regulation mechanism of the ovariectomized and preventive administration. In the femur and tibia of the rats, the fracture of the trabecular bone, the spacing and the disorder of the structure were observed. The prophylactic injection of beta estradiol (beta -estradiol-treated OVX, OVX/E2) after the ovariectomy had no obvious pathological symptoms. Compared to the sham operation (Sham-operated, Sham) group, the duodenum, the renal cortex and the renal cortex of group OVX and OVX/E2 group. There were no obvious lesions in the medullary region. Compared to group Sham, the number of uterine glands in the lamina propria of group OVX increased, the gland cavity increased, and the epithelium of the uterus was thickened in.OVX/E2 group, although the adeno gland of the uterus increased slightly, but the shape of the gland was more regular, and the endometrium had no obvious thickening. The amount of beta estradiol can prevent the occurrence of osteoporosis, and there is no.ELISA detection of the phenomenon of uterine carcinogenesis. The results of serum estrogen in the three different treatment groups showed no significant difference in serum estrogen. The results of.Real-time qPCR detection showed that the amount of PTH in the ovariectomized rats was significantly down, while the prophylactic injection of beta female was a prophylactic injection. After alcohol, the expression of PTH was significantly up-regulated in parathyroid glands, and the expression of ER- alpha was up-regulated after ovariectomized, while the expression of ER- alpha was significantly downregulated after the prophylactic injection of beta estradiol. It indicated that ER- alpha in parathyroid glands could respond to the weak changes in estrogen and participate in the expression and regulation of PTH. The expression of VDR in parathyroid glands of ovariectomized rats The amount of VDR was down regulated in the parathyroid parathyroid after prophylactic injection of beta estradiol, indicating that VDR may be the expression of inhibition of PTH. The serum Ca~ (2+) increased slightly after ovariectomized, and the expression of CaSR in parathyroid glands decreased significantly. The Ca~ (2+) increased slightly after the prophylactic injection of beta estradiol, and the expression of CaSR in parathyroid glands was significantly higher. The Ca~ (2+) /CaSR regulatory system may be involved in the regulation of PTH expression. The expression of FGF-23, FGFR1, Klotho and Egr1 in the parathyroid glands of the ovariectomized rats is significantly down, while the prophylactic injection of beta estradiol is down to FGF-23, Klotho and Egr1, and the expression of FGFR1 is up regulated, indicating the line in the parathyroid gland after ovariectomized. The inhibition of PTH expression is reduced, but the function of the receptor system after a prophylactic injection of beta estradiol is lower. These results suggest that ovariectomy and preventive administration may affect the ER- alpha, the VDR signal regulation system, the Ca~ (2+) /CaSR regulatory system and the FGF-23/Klotho receptor regulation system, and then regulate the table of PTH. Da.
【學(xué)位授予單位】：陜西理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R580
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