本文選題：甲狀旁腺素 + 信號轉導通路　； 參考：《南方醫(yī)科大學》2015年碩士論文

【摘要】：背景：甲狀旁腺素(PTH)是目前應用于臨床的唯一促骨形成藥物,它可以有效增加骨量,治療骨質疏松及骨質疏松性骨折、關節(jié)假體松動及長期使用二磷酸鹽引起的非典型骨折等。2002年美國FDA批準低劑量PTH間斷性皮下注射用以治療骨質疏松,2010年PTH被批準進入中國,治療絕經后的重度骨質疏松。隨著中國人口老齡化問題的日益嚴重,以及人們對于預防骨質疏松的意識尚淺薄等原因,骨質疏松癥愈來愈成為困擾中老年人的疾病之一。骨質疏松癥是一種以骨量減少及骨組織顯微結構退化為特征,導致骨脆性增加及骨折危險性增加的一種全身代謝性骨病,目前骨質疏松癥的治療藥物種類很多,其適用范圍和作用機制也各不相同。國內的治療主要以抗骨質吸收類藥物為主,包括二膦酸鹽類、降鈣素、雌激素及選擇性雌激素受體調節(jié)劑等,輔以促進骨礦化的鈣劑、維生素D及其活性代謝物等,其對于骨質疏松癥的治療效果有限。PTH作為一種骨形成促進劑,研究證明其對骨密度提高的程度優(yōu)于既往抗骨吸收類藥物,對減少骨質疏松性骨折的作用也更為突出。但是,PTH在臨床上的應用也受到諸多限制,如價格高,療程長(2年),高劑量、持續(xù)應用會促進骨吸收(不能復合到內植入物以促進骨融合),可能存在致癌的風險,長期使用還存在療效降低的可能性等,因此優(yōu)化PTH,促進療效,縮短使用時間,避免其副作用,是目前PTH領域研究的重要方向之一。人體中天然的PTH由84個氨基酸構成[PTH(1-84)],與Ⅰ型PTH受體(PTHR1)結合后激活PTHR1及其下游的多個信號途徑,參與骨組織代謝的過程。主要的信號通路有：(1) Gs/cAMP/PKA信號轉導通路,目前認為是PTH作用于骨組織的主要機制。(2) PLC/PKC通路。(3)非PLC依賴PKC激活途徑(PTH/nonPLC/PKC)。(4) β-arrestin通路等。目前認為,PTHR1受體至少通過兩種途徑激活PKC,其一為激活質膜上的PLC,進而激活PKC；另一途徑為不依賴PLC的PKC激活途徑(nonPLC/PKC),即不激活PLC而通過其他分子介導機制激活PKC,具體信號特征并不清楚,其信號轉導機制及調節(jié)骨代謝功能的研究是時下PTH研究領域的熱點之一。PTH激活不同的信號通路與其自身的多肽結構密不可分,任何氨基酸的突變及構象的改變都有可能導致某些信號通路激活能力的缺失。研究證實,改變PTH的氨基酸序列能改變PTH的信號轉導特征,如PTH(1-34)片段中,1-3位的氨基酸決定PTH的cAMP/PKA和PLC信號通路的激活功能；Ser1變成Gly1可使得PTH(1-34)失去激活PLC的能力；第19位氨基酸Glu19變成Arg19能夠補償29-34氨基酸殘基缺失引起的受體結合力減弱；Leu24,Leu28,va131突變成Glu后,PTH(5-34)不與受體結合Aib1,3,Nle8,21,Gln10,Har11,Ala12,Trp14,Arg19構成的組合突變亦稱M突變,M突變可使PTH(1-34)的cAMP合成能力提高約40倍,PLC的激活能力提高66倍,與PTHR的結合力提高約10倍；在PTH(1-34)中,Ile5,Glul9,Val21等位點的突變嚴重干擾PTH(1-34)與受體的結合力：PTH(1-34)第1、3位氨基酸突變成氨基異丁酸(Aib)后促進與PTHR的結合和cAMP的形成。根據以上模擬肽結構的研究基礎,我們擬通過氨基酸突變的方法來達到基本不影響PTH肽與受體結合能力,屏蔽cAMP/PKA和PLC激活能力的目的,設計nonPLC/PKC通路特異性PTH模擬肽的序列。我們的前期研究也表明,Gly1Arg19hPTH(1-28) (GR(1-28)和Gly1Arg19hPTH(1-34) (GR(1-34))均失去了激活PLC的能力,但GR(1-34)仍可激活PKC,而GR(1-28)卻沒有這樣的作用,即PTH可以通過nonPLC/PKC通路激活PKC,且功能區(qū)域是PTH(29-34)片段。通過C57BL小鼠皮下注射觀察發(fā)現,GR(1-34)顯著增加骨量,尤其是受力的松質骨區(qū)域,顯著強于GR(1-28)的作用,盡管后者具有與前者相同的cAMP激活特性,提示PTH作用下nonPLC/PKC通路的激活能夠促進受力區(qū)松質骨合成,改善骨小梁微結構,促進新骨的形成。近期我們發(fā)現GR(1-34)具有比PTH(1-34)和GR(1-28)更強的促進脊柱融合的作用,具有一定通路選擇性的PTH模擬肽具有較PTH更好的促骨形成作用和改善骨微結構的功能。但是nonPLC/PKC通路的具體信號介導分子仍不清楚,其對于骨代謝的作用機制仍需要深入研究。因此我們擬設計和篩選nonPLC/PKC信號通路選擇性PTH模擬肽,并分析該通路特有的下游效應分子,進一步探討PTH通過nonPLC/PKC信號通路促進骨形成的作用機理。目的：利用原代成骨細胞,通過相關信號通路屏蔽和基因表達分析,篩選與核實甲狀旁腺激素29-34位蛋白結構域(PTH(29-34))的效應基因,分析其對骨代謝的影響。以PTH結構為模板,通過改變其氨基酸序列,構建nonPLC/PKC信號通路選擇性PTH模擬肽,并在相關通路阻滯劑與激活劑的干擾下利用FRET技術和ELISA法對其信號特征進行驗證核實。并研究新肽對成骨相關基因的影響,探究PTH/nonPLC/PKC通路的骨代謝功能。方法：2-3日齡C57BL乳鼠10只,取顱蓋骨分離培養(yǎng)成骨細胞,經成骨誘導液培養(yǎng)14d和28d時分別進行ALP染色和茜素紅染色,鑒定原代成骨細胞。取貼壁生長的第1代細胞,分別接受100 nmol/L GR(1-28),10 nmol/L GR(1-34),10 nmol/L PTH(1-34)及空白對照作用4h,提取總RNA,行小鼠全基因組表達譜芯片分析,進行相關通路分析,并篩選出可能與nonPLC/PKC信號轉導通路相關的差異表達基因。RT-PCR篩選及驗證上述差異表達基因。培養(yǎng)MC3T3-E1細胞,使用cAMP通路抑制劑(RP-cAMP)阻斷PTH誘發(fā)的cAMP/PKA信號通路,比較GR(1-28)和GR(1-34)引起的基因變化情況。構建PTHR穩(wěn)定轉染的HEK293細胞,轉染CKAR報告分子后,利用PKC激活的熒光能量共振轉移(FRET)技術,檢測PTHR穩(wěn)轉細胞在cAMP通路抑制劑(RP-cAMP)干擾下分別接受GR(1-28)和GR(1-34)刺激下PKC的激活情況,確定PTH的nonPLC/PKC通路相關區(qū)域。以PTH氨基酸為基礎,用氨基酸突變的方法增強nonPLC/PKC通路相關區(qū)域作用,合成PKA"PLCPKC+肽(MY1肽)。用FRET技術檢測MYl肽對轉染CKAR的PTHR穩(wěn)轉HEK293細胞中PKC的激活能力,并觀察加入PKC抑制劑(Go6983)后PKC激活的變化情況,檢測MY1肽作用下轉染CKAR的HEK293細胞(無PTHR)中PKC的激活情況,探究其與PTHR的關系。用ELISA法檢測MY1肽對MC3T3-E1細胞中cAMP、PLC激活能力。以驗證核實MYl肽為PTH的nonPLC/PKC通路特異性模擬肽。MC3T3-E1細胞分別接受10umol/LMY1肽、10umol/L PTH(3-34、100nmol/LPTH(1-34)及空白對照組作用4h,提取總RNA, RT-PCR檢測成骨相關基因的表達。結果：原代培養(yǎng)成骨細胞細胞形態(tài)較均一,呈梭形或多邊形。待細胞長滿后,見細胞排列緊密,呈鋪路石狀。成骨誘導培養(yǎng)14d,細胞排列緊密,呈復層生長,行ALP染色,鏡下顯示胞質中出現藍色顆粒,成骨誘導培養(yǎng)至28d行茜素紅染色,鏡下顯示出現紅染的礦化結節(jié)。行小鼠全基因組芯片檢測,通過相關通路分析,得到了最可能與PTH的nonPLC/PKC信號轉導途徑相關的14條信號通路。根據芯片結果,經過進一步分析,我們挑選出了與PTH的nonPLC/PKC信號通路相關性最高的56個基因,作為RT-PCR篩選驗證的基因對象。篩選的56個基因中,我們發(fā)現CITED1的表達量PTH(1-34)組明顯高于其他各組,且GR(1-34)組顯著高于GR(1-28)組,PTH(1-34)與GR(1-34)組均顯著高于空白對照組。MC3T3-E1細胞經各組模擬肽刺激后,提取總RNA,進行RT-PCR檢測,CITED1的表達量GR(1-34)組仍顯著高于GR(1-28)組,與原代成骨細胞實驗結果一致,且在加入PKC抑制劑后,CITED1表達量明顯下降,證實CITED1表達量的升高由nonPLC/PKC通道激活引起,不依賴PLC及PKA的激活。PTHR穩(wěn)轉細胞經PKC特異性激活劑TPA刺激后,PKC被激活,C/Y值顯著增高,表明我們檢測PKC激活的FRET技術檢測平臺構建成功,其可以作為后續(xù)試驗中檢測PKC激活的有效手段。PTHR穩(wěn)轉細胞在PTH(1-34)的作用下,C/Y值顯著增高,即檢測到PKC被激活,表明我們成功地構建了PTHR穩(wěn)轉細胞。在轉染CKAR的PTHR穩(wěn)轉細胞細胞中,阻斷cAMP通路后,GR(1-28)無法激活PKC,而GR(1-34)仍可激活PKC,nonPLC/PKC通路的激活與PTH的29至34氨基酸片段有關。在MYl肽作用下,FRET檢測顯示C/Y值明顯升高,PKC被激活,而PTH(3-34)及空白對照中,PKC未被激活。在MYl肽刺激使C/Y值明顯升高后,加入PKC阻滯劑后,C/Y值明顯下降,并逐漸降至起始水平。轉染CKAR的HEK293細胞(無PTHR)在PTH(3-34)或是MYl作用下,C/Y值均未發(fā)生變化,而在加入TPA (PKC特異性激活劑)后,C/Y值明顯升高,PKC被激活。ELISA法檢測MC3T3-E1經各模擬肽作用下cAMP、PLC的含量,實驗結果顯示,PTH(1-34)組中cAMP、PLC含量明顯高于其它各組,而其它三組之間cAMP含量無統(tǒng)計學差異,即MYl肽沒有激活cAMP、PLC的能力。MC3T3-E1細胞經各PTH模擬肽作用后行RT-PCR檢測,結果顯示MYl組CITED1的表達量明顯高于空白對照組和PTH(3-34)組,在加入PKC抑制劑Go6983后,CITED1的表達量明顯下降,與原代成骨細胞實驗中結果相一致。在成骨基因檢測中,ALP的表達量在MYl組明顯高于空白對照組和PTH(3-34)組,且在加入PKC抑制劑后,其表達量明顯下降。結論：1.原代成骨細胞培養(yǎng)成功,經基因芯片分析,發(fā)現與PTH的nonPLC/PKC信號轉導途徑相關的14條信號通路。經RT-PCR篩選,我們發(fā)現PTH(29-34)蛋白機構域可通過PKC信號途徑促進CITED1的表達,介導PTH對成骨代謝的作用。該途徑不依賴PLC和PKA信號的激活。2.經FRET分析,PTHR穩(wěn)定轉染細胞成功建立,我們構建的MY1肽具有通過PTHR激活PKC的特性,且不依賴PKA及PLC信號轉導。表明MY1為PTH/nonPLC/PKC通路特異性模擬肽。MYl肽的建立為PTH/nonPLC/PKC的進一步研究奠定了基礎,據我們所知,單純具有nonPLC/PKC的信號特征的PTH模擬肽國內外未有報道。3.MYl肽促進轉錄因子CITED1和成骨基因ALP的表達,表明PTH的nonPLC/PKC通路至少通過調控轉錄因子CITED1或成骨基因ALP的表達量參與骨代謝調節(jié)過程。但其對骨代謝的影響和機理需要進一步的研究。
[Abstract]:Background: parathyroid hormone (PTH) is the only bone forming drug used in clinical practice. It can effectively increase bone mass, treat osteoporosis and osteoporotic fractures, joint prosthesis loosening and atypical fractures caused by long-term use of two phosphate and other.2002 years, FDA batch low dose PTH intermittent hypodermic injection for the treatment of bone Loosely, in 2010, PTH was approved to enter China for the treatment of postmenopausal severe osteoporosis. Osteoporosis is becoming one of the diseases that plagued middle and old people as the problem of aging population in China is increasingly serious and the awareness of osteoporosis is still shallow. Osteoporosis is a kind of bone loss and bone. A kind of systemic metabolic bone disease characterized by the degeneration of microstructure and the increase of bone fragility and increased risk of fracture. There are many kinds of drugs for the treatment of osteoporosis, and the scope and mechanism of its application are different. The main treatment in China is anti bone absorption drugs, including two phosphonic acid salts, calcitonin, and female irritable disease. Hormone and selective estrogen receptor modulators, supplemented with calcium, vitamin D and its active metabolites that promote bone mineralization, and its therapeutic effect on osteoporosis is limited.PTH as a bone formation promoter. Studies have shown that its increase in bone density is better than previous anti bone resorption drugs for reducing osteoporotic fractures. However, the clinical application of PTH is also limited, such as high price, long course (2 years), high dose, continuous application will promote bone absorption, which can not be combined with internal implant to promote bone fusion. It may have the risk of carcinogenesis, and the long-term use still has the possibility of reducing the curative effect, so optimizing the PTH and promoting the curative effect, Shortening the use time and avoiding its side effects are one of the most important directions in the field of PTH. The natural PTH in the human body consists of 84 amino acids [PTH (1-84)], which is combined with the type I PTH receptor (PTHR1) to activate PTHR1 and its downstream signal pathway to participate in the metabolic process of bone tissue. The main signal pathways are: (1) Gs/cAMP/PKA signal Transduction pathway is considered to be the main mechanism that PTH acts on bone tissue. (2) PLC/PKC pathway. (3) non PLC dependent PKC activation pathway (PTH/nonPLC/PKC). (4) beta -arrestin pathway, etc. at present, the PTHR1 receptor activates at least two ways to activate PKC, one is activating the PLC on the plasma membrane and activating PKC; the other is not dependent on PLC PKC excitation. The active pathway (nonPLC/PKC), which is not activated by PLC, activates PKC through other molecular mediated mechanisms. The specific signal characteristics are not clear. The signal transduction mechanism and the regulation of bone metabolism are one of the hotspots in the current field of PTH research..PTH activation of different signaling pathways is inseparable from its own peptide structure and any amino acid mutation. Changes in conformation and conformation may lead to loss of activation ability of certain signal pathways. Studies have shown that changing the amino acid sequence of PTH can change the signal transduction characteristics of PTH, such as PTH (1-34) fragments, the 1-3 amino acids determine the activation function of the cAMP/PKA and PLC signaling pathways of PTH; Ser1 becomes Gly1 can cause PTH (1-34) to lose activation of PLC. Nineteenth bit amino acid Glu19 Arg19 can compensate for the weakening of the receptor binding force caused by the deletion of the 29-34 amino acid residues; after Leu24, Leu28, and va131 process into Glu, PTH (5-34) does not combine Aib1,3 with the receptor, Nle8,21, Gln10, Har11, Ala12, and is also called the mutation. 40 times, the activation ability of PLC increased by 66 times and the binding power of PTHR increased by about 10 times; in PTH (1-34), the mutation of Ile5, Glul9, Val21 and other sites seriously interfered with the binding force of PTH (1-34) with the receptor: PTH (1-34) 1,3 amino acid suddenly became the combination of amino isobutyric acid (Aib) and PTHR binding and cAMP formation. On the basis of this, we intend to design the sequence of the nonPLC/PKC pathway specific PTH analog peptide by means of amino acid mutation, which basically does not affect the binding ability of PTH peptide to the receptor, shielding the activation ability of cAMP/PKA and PLC. Our previous study also showed that Gly1Arg19hPTH (1-28) (GR (1-28) and Gly1Arg19hPTH (1-34) (GR (1-34)) were lost. The ability to activate PLC, but GR (1-34) still activates PKC, while GR (1-28) does not function, that is, PTH can activate PKC through the nonPLC/PKC pathway, and the functional region is PTH (29-34). By subcutaneous injection of the C57BL mice, it is found that GR (1-34) significantly increases the bone mass, especially the region of the stressed cancellous bone, which is significantly stronger than GR (1-28). Although the latter has the same cAMP activation characteristics as the former, it is suggested that activation of the nonPLC/PKC pathway under the action of PTH can promote the synthesis of cancellous bone in the stressed area, improve the microstructure of the trabecular bone and promote the formation of new bone. In the near future, we found that GR (1-34) has a stronger role in promoting spinal fusion than PTH (1-34) and GR (1-28), and has a certain pathway selectivity. The PTH mimic peptide has a better function of promoting bone formation and improving bone microstructure than PTH. However, the specific signaling molecules of the nonPLC/PKC pathway are still unclear, and the mechanism of its action on bone metabolism still needs to be studied. Therefore, we intend to design and screen the selective PTH analog peptide of the nonPLC/PKC signaling pathway and analyze the pathway of this pathway. Some downstream effectors further explore the mechanism of PTH to promote bone formation through the nonPLC/PKC signaling pathway. Objective: to screen and verify the effect genes of parathyroid hormone 29-34 protein domain (PTH (29-34)) by using primary osteoblasts, screening and verifying the gene expression domain (PTH (29-34)) of parathyroid hormone, and analyzing its effect on bone metabolism. With the PTH structure as a template, the nonPLC/PKC signaling pathway selective PTH mimic peptide was constructed by changing the amino acid sequence, and the signal characteristics were verified by FRET and ELISA under the interference of the related pathway blockers and activators, and the effect of the new peptide on the osteogenic phase gene was investigated and the PTH/nonPLC/PKC pathway was explored. Methods: bone metabolism function. Methods: 10 C57BL mice aged 2-3 days old were isolated and cultured for osteoblasts. ALP staining and alizarin red staining were carried out when 14d and 28d were cultured by osteogenic induction solution to identify the primary osteoblasts. The first generation cells attached to the wall were taken to receive 100 nmol/L GR (1-28), 10 nmol/L GR (1-34), 10 nmol/L PTH (1-34) and empty, respectively. The total RNA was extracted from the white control 4h, the whole genome expression spectrum chip was analyzed, the related pathway was analyzed, and the differential expression gene.RT-PCR related to the nonPLC/PKC signal transduction pathway was screened and tested to verify the differential expression genes. MC3T3-E1 cells were cultured and cAMP pathway inhibitor (RP-cAMP) was used to block cAMP induced cAMP. /PKA signaling pathway, comparing the gene changes caused by GR (1-28) and GR (1-34). Construction of PTHR stable transfected HEK293 cells, after transfection of CKAR reporter molecules, using PKC activated fluorescence energy resonance transfer (FRET) technology to detect the PTHR metastable cells under cAMP pathway inhibitor (RP-cAMP) and receive GR (1-28) and stimulus (1-34) stimulation respectively. Activation and determination of the nonPLC/PKC pathway related regions of PTH. Based on PTH amino acids, the function of nonPLC/PKC pathway related regions was enhanced by amino acid mutation, and PKA "PLCPKC+ peptide (MY1 peptide) was synthesized. The activation ability of MYl peptide to PTHR stabilized HEK293 cells transfected with CKAR was detected by FRET technology. The activation of PKC was detected by detecting the activation of PKC in HEK293 cells transfected with CKAR under the action of MY1 peptide (PTHR), and the relationship between them and PTHR was explored. ELISA assay was used to detect cAMP and PLC activation of MY1 peptides in MC3T3-E1 cells. 1 0umol/L PTH (3-34100nmol/LPTH (1-34) and blank control group acted 4h, extracted total RNA and RT-PCR to detect the expression of bone related genes. Results: the cells of primary cultured osteoblasts were homogenous, shuttle or polygon. After the cell was full, the cells were arranged closely and showed a paved stone. The osteogenesis was induced and cultured for 14d, the cells were arranged close and complex. The layer was grown and stained with ALP. The blue particles appeared in the cytoplasm under the microscope. The osteogenesis was induced to 28d with alizarin red staining. The red stained mineralized nodules were displayed under the microscope. All the 14 signal pathways related to the nonPLC/PKC signal transduction pathway of PTH were obtained by the whole genome chip detection. As a result, after further analysis, we selected 56 genes with the highest correlation with the nonPLC/PKC signaling pathway of PTH, which were selected as RT-PCR screening tests. Among the 56 selected genes, we found that the PTH (1-34) group of CITED1 was significantly higher than the other groups, and GR (1-34) group was significantly higher than the GR (1-28) group, PTH (1-34) and GR (1-34). The group of.MC3T3-E1 cells was significantly higher than that of the blank control group. After the stimulation of the analog peptide, the total RNA was extracted and the RT-PCR was detected. The expression of CITED1 was still significantly higher than that in the GR (1-28) group, which was consistent with the experimental results of the original osteoblast, and the expression of CITED1 was obviously decreased after adding the PKC inhibitor. The increase of the CITED1 expression was confirmed by nonPLC. The increase of the CITED1 expression was confirmed by nonPLC. The activation of /PKC channel, which does not depend on the activation of PLC and PKA, is activated by PKC specific activator TPA, PKC is activated and C/Y value increases significantly. It indicates that our detection platform for FRET is successful. It can be used as an effective means to detect PKC excitation in the follow-up test. At the same time, the C/Y value increased significantly, that is, the detection of PKC was activated, indicating that we successfully constructed the PTHR stable cells. In the PTHR stable transfected cell cells transfected with CKAR, GR (1-28) can not activate PKC, and GR (1-34) still activates PKC. The activation of the nonPLC/PKC pathway is related to the 29 to 34 amino acid fragments of PTH. The test showed that the C/Y value was obviously increased and PKC was activated, while PKC was not activated in PTH (3-34) and blank control. After the MYl peptide stimulated the C/Y value, the C/Y value decreased obviously, and gradually decreased to the beginning level. The HEK293 cells transfected with CKAR were not changed in PTH (3-34) or under the action. After TPA (PKC specific activator), the C/Y value increased obviously. PKC was activated by.ELISA method to detect the content of cAMP and PLC under the action of each analogue peptide of MC3T3-E1. The experimental results showed that the PLC content in PTH (1-34) group was obviously higher than that of the other groups, but the cAMP content of the other three groups was not statistically different. The expression of CITED1 in MYl group was significantly higher than that of the blank control group and PTH (3-34) group. The expression of CITED1 in the MYl group was significantly higher than that in the blank control group and the PTH (3-34) group. The expression of ALP was significantly higher in the MYl group than in the original osteoblast test. In the blank control group and the PTH (3-34) group, and after the addition of PKC inhibitor, the expression of the 1. primary osteoblasts was successfully cultured. The 14 signal pathways associated with the nonPLC/PKC signal transduction pathway of PTH were detected by gene chip analysis. We found that the PTH (29-34) protein mechanism domain could be promoted by the PKC signal pathway through the RT-PCR screening. The expression of CITED1 mediates the effect of PTH on osteoblastic metabolism. This pathway does not depend on the activation of PLC and PKA signals by FRET analysis, and PTHR stable transfection cells are successfully established. The MY1 peptide we constructed has the characteristics of activating PKC through PTHR, and does not depend on PKA and PLC signal transduction. The foundation is established for further study of PTH/nonPLC/PKC. As we know, the PTH analog peptide with nonPLC/PKC signal characteristics has not reported the expression of.3.MYl peptide promoting transcription factor CITED1 and osteogenic gene ALP, indicating that the nonPLC/PKC pathway of PTH is at least by regulating the expression of the transcription factor CITED1 or the expression of ALP of the osteogenic gene. Volume is involved in the process of bone metabolism regulation, but its effects on bone metabolism and mechanism need further study.
【學位授予單位】：南方醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R580

【共引文獻】

相關期刊論文 前2條

1 郝松;孟越;李威;胡少宇;楊德鴻;;甲狀旁腺素非PLC依賴PKC通路激活增強成骨細胞CITED1表達[J];南方醫(yī)科大學學報;2015年04期

2 吳素珍;李加林;;甲狀旁腺激素相關蛋白研究進展[J];中國生化藥物雜志;2015年07期

相關博士學位論文 前3條

1 耿文鑫;抗絕經后骨質疏松蛋白疫苗的實驗研究[D];西北大學;2013年

2 趙晶;北京地區(qū)漢族絕經后婦女MATN3基因多態(tài)性與骨質疏松表型的關聯研究[D];北京協(xié)和醫(yī)學院;2010年

3 劉雅紅;甲狀旁腺素相關肽核定位序列和C末端在調節(jié)中樞神經系統(tǒng)發(fā)育中的作用及機制研究[D];南京醫(yī)科大學;2013年

相關碩士學位論文 前5條

1 代守前;血清素對成骨細胞增殖、分化和礦化功能及對小鼠骨質參數的影響[D];南京醫(yī)科大學;2013年

2 袁亮;信號選擇性甲狀旁腺素模擬肽影響去勢雄性小鼠骨折愈合的實驗研究[D];南方醫(yī)科大學;2013年

3 李振志;杜仲籽提取物對去勢大鼠骨質疏松的作用機制研究[D];西北大學;2014年

4 孟越;甲狀旁腺素通過PKA依賴及非依賴途徑激活PKC信號通路的實驗研究[D];南方醫(yī)科大學;2014年

5 王金鳳;內源性FGF23對PTH促成骨細胞分化作用的影響[D];復旦大學;2013年

，

本文編號：1982720