Exogenous bone morphogenetic protein-7 reduces hepatic fibrosis inSchistosoma japonicum-infected micevia transforming growth factor-β/Smad signaling

AIM: To investigate the antifibrotic effects of bone morphogenetic protein-7 (BMP-7) on Schistosoma japonicum (S. japonicum )-induced hepatic fibrosis in BALB/C mice. METHODS: Sixty BALB/C mice were randomly divided into three groups, including a control group (group A, n = 20), model group (group B, n = 20) and BMP-7 treated group (group C, n = 20). The mice in group B and group C were abdominally infected with S. japonicum cercariae to induce a schistosomal hepatic fibrosis model. The mice in group C were administered human recombinant BMP-7. Liver samples were extracted from mice sacrificed at 9 and 15 wk after modeling. Hepatic histopathological changes were assessed using Masson's staining. Transforming growth factor-beta 1 (TGF-β1), alpha-smooth muscle actin (α-SMA), phosphorylated Smad2/3 (pSmad2/3) and Smad7 protein levels and localization were measured by Western blotting and immunohistochemistry, respectively, and their mRNA expressions were detected by reverse transcriptionpolymerase chain reaction (RT-PCR). RESULTS: The schistosomal hepatic fibrosis mouse model was successfully established, as the livers of mice in group B and group C showed varying degrees of typical schistosomal hepatopathologic changes such as egg granuloma and collagen deposition. The degree of collagen deposition in group C was higher than that in group A (week 9: 22.95±6.66vs 2.02±0.76; week 15: 12.84±4.36 vs 1.74±0.80; P<0.05), but significantly lower than that in group B (week 9: 22.95±6.66 vs 34.43±6.96; week 15: 12.84±4.36 vs 18.90±5.07;P<0.05) at both time points. According to immunohistochemistry data, the expressions of α-SMA, TGF-β1 and pSmad2/3 protein in group C were higher than those in group A (α-SMA: week 9: 21.24±5.73 vs 0.33±0.20; week 15: 12.42±4.88 vs 0.34±0.27; TGF-β1: week 9: 37.00±13.74 vs 3.73±2.14; week 15: 16.71±9.80 vs 3.08±2.35; pSmad2/3: week 9: 12.92±4.81 vs 0.83±0.48; week 15: 7.87±4.09 vs 0.90±0.45; P<0.05), but significantly lower than those in group B (α-SMA: week 9: 21.24±5.73 vs 34.39±5.74; week 15: 12.42±4.88 vs 25.90±7.01; TGF-β1: week 9: 37.00±13.74 vs 55.66±14.88; week 15: 16.71±9.80 vs 37.10±12.51; pSmad2/3: week 9: 12.92±4.81 vs 19.41±6.87; week 15: 7.87±4.09vs 13.00±4.98;P<0.05) at both time points; the expression of Smad7 protein in group B was higher than that in group A and group C at week 9 (8.46±3.95 vs 1.00±0.40 and 8.46±3.95 vs 0.77±0.42; P<0.05), while there were no differences in Smad7 expression between the three groups at week 15 (1.09±0.38 vs 0.97±0.42 vs 0.89±0.39; P>0.05). Although minor discrepancies were observed, the results of RT-PCR and Western blotting were mainly consistentwith the immunohistochemical results. CONCLUSION: Exogenous BMP-7 significantly decreased the degree of hepatic fibrosis in both the acute and chronic stages of hepato-schistosomiasis, and the regulatory mechanism may involve the TGF-β/Smad signaling pathway.

Heterotopic ossification after the use of recombinant human bone morphogenetic protein-7

AIM To present the incidence of heterotopic ossification after the use of recombinant human bone morphogenetic protein-7(rhB MP-7) for the treatment of nonunions.METHODS Bone morphogenetic proteins(BMPs) promote bone formation by auto-induction. Recombinant human BMP-7 in combination with bone grafts was used in 84 patients for the treatment of long bone nonunions. All patients were evaluated radiographicaly for the development of heterotopic ossification during the standard assessment for the nonunion healing. In all patients(80.9%) with radiographic signs of heterotopic ossification, a CT scan was performed. Nonunion site palpation and ROM evaluation of the adjacent jointswere also carried out. Factors related to the patient(age, gender), the nonunion(location, size, chronicity, number of previous procedures, infection, surrounding tissues condition) and the surgical procedure(graft and fixation type, amount of rhB MP-7) were correlated with the development of heterotopic ossification and statistical analysis with Pearsons χ~2 test was performed.RESULTS Eighty point nine percent of the nonunions treated with rh BMP-7, healed with no need for further procedures. Heterotopic bone formation occurred in 15 of 84 patients(17.8%) and it was apparent in the routine radiologi-cal evaluation of the nonunion site, in a mean time of 5.5 mo after the rh BMP-7 application(range 3-12). The heterotopic ossification was located at the femur in 8 cases, at the tibia in 6, and at the humerus in οne patient. In 4 patients a palpable mass was present and only in one patient, with a para-articular knee nonunion treated with rhB MP-7, the size of heterotopic ossification affected the knee range of motion. All the patients with heterotopic ossification were male. Statistical analysis proved that patient's gender was the only important factor for the development of heterotopic ossification(P = 0.007). CONCLUSION Heterotopic ossification after the use of rh BMP-7 in nonunions was common but it did not compromise the final clinical outcome in most cases, and affected only male patients.

RETINOIC ACID DOWN-REGULATES BONE MORPHOGENETIC PROTEIN 7 EXPRESSION IN RAT WITH CLEFT PALATE

Objective To evaluate the effects of retinoic acid （RA） on expression of bone morphogenetic protein 7 （ BMP-7 ） in rat fetus with cleft palate, and the effects of RA on proliferation and apoptosis of osteoblasts. Methods All-trans RA （ATRA） was used to induce congenital cleft palate in Wistar rat. BMP-7 mRNA expression in maxillary bone tissue of fetal rats was measured by Northern blotting analysis. Flow cytometry and MTF assay were used to measure the apoptosis and proliferation of ATRA-treated MC-3T3-E1 cells. BMP-7 mRNA and protein expressions in ATRA-treated MC-3T3-E1 cells were detected by RT-PCR and Western blotting analysis. Remilts ATRA could induce cleft palate of rat fetus. The incidence rate of cleft palate induced by 100 mg/kg AT-RA （45.5%） was significantly higher than 50 mg/kg ATRA （ 12.5%, P 〈 0. 05 ）. BMP-7 mRNA expression decreased in maxillary bone tissue of rat fetus with cleft palate. MC-3T3-E1 cells proliferation treated with 1 × 10^-6 mol/L ATRA decreased by 60%, the cell apoptosis increased by 2 times. BMP-7 mRNA and protein levels in MC-3T3-E1 cells treated with 1 × 10^-6 mol/L ATRA decreased by 60% and 80%, respectively, compared with ATRA-untreated cells （ P 〈 0.05 ）. Conclusions BMP-7 may play an important role in embryonic palate development. RA may possess the ability to down-regulate cell proliferation through regulation of BMP-7 gene expression.

Roles and regulation of bone morphogenetic protein-7 in kidney development and diseases

The gene encoding bone morphogenetic protein-7(BMP7) is expressed in the developing kidney in embryos and also in the mature organ in adults. During kidney development, expression of BMP7 is essential to determine the final number of nephrons in and proper size of the organ. The secreted BMP7 acts on the nephron progenitor cells to exert its dual functions: To maintain and expand the progenitor population and to provide them with competence to respond to differentiation cues, each relying on distinct signaling pathways. Intriguingly, in the adult organ, BMP7 has been implicated in protection against and regeneration from injury. Exogenous administration of recombinant BMP7 to animal models of kidney diseases has shown promising effects in counteracting inflammation, apoptosis and fibrosis evoked upon injury. Although the expression pattern of BMP7 has been well described, the mechanisms by which it is regulated have remained elusive and the processes by which the secretion sites of BMP7 impinge upon its functions in kidney development and diseases have not yet been assessed. Understanding the regulatory mechanisms will pave the way towards gaining better insight into the roles of BMP7, and to achieving desired control of the gene expression as a therapeutic strategy for kidney diseases.

Construction of Adeno-associated Virus System for Human Bone Morphogenetic Protein 7 Gene

To construct the recombinant adeno-associated virus （rAAV） vector with human bone morphogenetic protein 7 （BMP7） and observe the BMP7 mRNA expression in vitro, BMP7 CDS sequence was cloned into expression plasmid pAAV-MCS of AAV Helper Free System. The recombinant plasmid was identified with enzyme digestion and sequencing. The recombinant plasmid, pAAV-RC, pHelper were co-transfected into AAV-293 cells according to the calcium phosphate-based protocol. The viral stock was collected by 4 rounds of freeze/thaw. After purified and concentrated, the recombinant virus titer was determined by dot-blot assay. HEK293 cells were transfected with the recombinant virus at different MOI, and the expression of BMP7 mRNA was detected by RT-PCR. The results showed rAAV-BMP7 was constructed and packaged successfully. The physical particle titer was 2.5×10^11 vector genomes/mL. There was different expression level of BMP7 mRNA after transfecton. These data suggested that recombinant AAV mediated a stable expression of hBMP7 mRNA in 293 cells. The AAV production method may pave the way of an effective strategy for the jaw bone defection around dental implants.

Bone morphogenetic protein-7 induced bone marrow stromal cells differentiate into neuron-like cells

Bone morphogenetic protein-7 is widely accepted as an inducer for bone marrow stem cells differentiating into osteoblasts and chondrocytes. Whether bone marrow stromal cells differentiate into neuron-like cells remains unclear. The current study examined the presence of positive cells for intermediate filament protein and microtubule associated protein-2 in the cytoplasm of bone marrow stromal cells induced by bone morphogenetic protein-7 under an inverted microscope, while no expression of glial fibrillary acidic protein was found. Reverse transcription PCR electrophoresis also revealed a positive target band for intermediate filament protein and microtubule-associated protein 2 mRNA. These results confirmed that bone morphogenetic protein-7 induces rat bone marrow stromal cells differentiating into neuron-like cells.

Expression of bone morphogenetic protein 7 in the cerebral cortex of rats after ischemic-hypoxic injury

BACKGROUND： Some researches demonstrate that exogenous bone morphogenetic protein 7 （BMP-7） can protect ischemic cerebral nerve tissue and promote recovery of motor energy function; however, there is lack of direct evidences of endogenous BMP-7 effect. OBJECTIVE： To observe the expression of endogenous BMP-7 in nerve tissue with ischemic-hypoxic injury and investigate the possible effects on damaged nerve tissue. DESIGN： Observational contrast animal study. SETTING： Department of Anatomy and Histoembryology, Peking University Health Science Center. MATERIALS： The experiment was carried out in the Nerve Researching Laboratory of Anatomy Department, Peking University Health Science Center from October 2006 to March 2007. A total of 25 adult male SD rats weighing 250 - 300 g and several newborn SD rats were selected from Experimental Animal Center, Peking University Health Science Center. Rabbit-anti-BMP-7 polyclonal antibody was provided by Wuhan Boster Company. METHODS： ① Adult rats were randomly divided into ischemia group （n =10）, sham operation group （n = 10） and normal group （n =5）. Right external-internal carotid artery occlusion was used to infarct middle cerebral artery of adult rats in the ischemia group so as to copy focal cerebral infarction models. Line cork was inserted in crotch of internal and external carotid artery of adult rats in the sham operation group, while adult rats in the normal group were not given any treatments. ② Cerebral cortex of newborn rats was separated to obtain cell suspension. Cells which were cultured for 10 days were divided into control group and hypoxia/reoxygenation group. And then, cells in the hypoxia/reoxygenation group were cultured in hypoxic incubator for 4 hours and given reoxygenation for 24 hours. MAIN OUTCOME MEASURES： Immunohistochemical method was used to measure expression of BMP-7 in cerebral cortex at 24 hours after ischemia/reperfusion culture and in primary hypoxic culture. RESULTS： ① At 24 hours after cerebral ischemia, expression of BMP-7 in cerebral cortex on ischemic side was stronger than that on non-ischemic side in adult rats; meanwhile, numbers of cell expression were increased. However, expression of BMP-7 was not detected in bilateral cerebral cortex of adult rats in both control group and sham operation group. ② After hypoxia of cerebral cortex in primary culture, positive products of BMP-7 were observed in plasma of neuron, but expression of BMP-7 was not found in normal cerebral cortex. CONCLUSION： Endogenous BMP-7 has protective effects on nerve tissue induced by ischemic-hypoxic injury.

基于BMP-2/BMP-7机制探讨生龙接骨胶囊预防老年骨质疏松性胸腰椎骨折术后再发性骨折的作用

目的探讨生龙接骨胶囊对老年骨质疏松性胸腰椎骨折(OTF)术后再发性骨折的作用,并基于骨形态发生蛋白-2/骨形态发生蛋白-7(BMP-2/BMP-7)机制初步分析其作用机制。方法选取2020年1月至2022年10月在南昌市洪都中医院收治的100例行经皮椎体成形术(PVP)的老年OTF患者为研究对象,按随机数字表法分为常规组(50例)和胶囊组(50例)。常规组采用常规的PVP治疗,胶囊组患者在常规组基础上服用生龙接骨胶囊治疗。比较两组患者椎体结构(Cobb角和伤椎椎体前缘高度比)、治疗前后血清BMP-2、BMP-7水平、骨代谢指标[骨特异性碱性磷酸酶(BALP)、Ⅰ型原胶原N端前肽(PⅠNP)、骨钙素(OST)]、骨密度(BMD)、康复情况[Oswestry腰椎功能障碍指数(ODI)]评估、疼痛等级[视觉模拟评分法(VAS)]评分、临床有效率、椎体再骨折发生率和不良反应发生情况。结果治疗后,两组患者Cobb角改善,伤椎椎体前缘高度比优于治疗前,胶囊组均优于常规组,差异有统计学意义(P<0.05);胶囊组的BMP-2、BMP-7表达量高于常规组,差异有统计学意义(P<0.05);患者术后再发性骨折情况均良好,胶囊组愈合速度快于常规组,差异有统计学意义(P<0.05);治疗后,胶囊组的BALP、PⅠNP、OST均高于常规组,差异有统计学意义(P<0.05);两组患者的BMD高于治疗前,且胶囊组高于常规组,ODI评分、VAS评分均低于治疗前,且胶囊组低于常规组,差异有统计学意义(P<0.05);胶囊组的临床总有效率(95.0%)高于常规组(80.0%),胶囊组的再骨折发生率(4%)低于常规组(18%),差异有统计学意义(P<0.05);两组患者均未见明显不良反应。结论生龙接骨胶囊能通过上调患者血清BMP-2、BMP-7水平,改善BMD从而预防老年OTF的术后再发性骨折,其机制可能与生龙接骨胶囊能刺激BMP信号通路,加速成骨细胞分化有关。

A型肉毒毒素防治额部外伤瘢痕的美学效果及对血清TGF-β_(1)、BMP-7水平的影响

目的:探讨A型肉毒毒素防治额部外伤瘢痕的美学效果及对血清TGF-β_(1)、BMP-7水平的影响。方法:选取2020年7月-2022年10月笔者医院收治的84例额部外伤患者为研究对象,按随机数字表法分为观察组和对照组,各42例。对照组采用清创美容缝合联合外用硅凝胶制剂防治瘢痕;观察组采用美容缝合拆线后伤口两侧注射A型肉毒毒素防治瘢痕。拆线后3个月,统计比较两组瘢痕临床防治有效率、瘢痕评分[温哥华瘢痕量表(Vancouver scar scale,VSS)]、瘢痕疼痛或瘙痒程度评分[视觉模拟评分法(Visual analogue scale,VAS)]、患者满意度、血清转化生长因子β_(1)(Transforming growth factor-β_(1),TGF-β_(1))和骨成型蛋白7(Recombinant bone morphogenetic protein 7,BMP-7)水平及不良反应。结果:观察组VSS评分、瘢痕疼痛或瘙痒程度VAS评分低于对照组(P<0.05);观察组瘢痕临床防治有效率为90.47%,高于对照组的69.04%(P<0.05);观察组患者满意度高于对照组(P<0.05);观察组TGF-β_(1)水平低于对照组,BMP-7水平高于对照组(P<0.05);两组均未发生严重不良反应。结论:额部软组织外伤患者美容缝合拆线后伤口两侧注射A型肉毒毒素可抑制瘢痕形成,有效提升瘢痕防治有效率及患者满意度,其临床效果可能与调控血清TGF-β_(1)和BMP-7水平有关,且安全性较高,具有一定的临床应用价值。

Bone morphogenetic protein 7 mediates stem cells migration and angiogenesis:therapeutic potential for endogenous pulp regeneration

Pulp loss is accompanied by the functional impairment of defense,sensory,and nutrition supply.The approach based on endogenous stem cells is a potential strategy for pulp regeneration.However,endogenous stem cell sources,exogenous regenerative signals,and neovascularization are major difficulties for pulp regeneration based on endogenous stem cells.Therefore,the purpose of our research is to seek an effective cytokines delivery strategy and bioactive materials to reestablish an ideal regenerative microenvironment for pulp regeneration.In in vitro study,we investigated the effects of Wnt3a,transforming growth factor-beta 1,and bone morphogenetic protein 7(BMP7)on human dental pulp stem cells(h-DPSCs)and human umbilical vein endothelial cells.2D and 3D culture systems based on collagen gel,matrigel,and gelatin methacryloyl were fabricated to evaluate the morphology and viability of h-DPSCs.In in vivo study,an ectopic nude mouse model and an in situ beagle dog model were established to investigate the possibility of pulp regeneration by implanting collagen gel loading BMP7.We concluded that BMP7promoted the migration and odontogenic differentiation of h-DPSCs and vessel formation.Collagen gel maintained the cell adhesion,cell spreading,and cell viability of h-DPSCs in 2D or 3D culture.The transplantation of collagen gel loading BMP7 induced vascularized pulp-like tissue regeneration in vivo.The injectable approach based on collagen gel loading BMP7 might exert promising therapeutic application in endogenous pulp regeneration.

Molecular Cloning and Sequence Analysis of FullLength cDNA Encoding Human Bone Morphogenetic Protein—7

Objective:To clone the full-length human bane morphogenetic protein-7 (BMP-7 ) gene and analyse its sequence, to aid in investigation of its function and structure. Methods : Total RNA was isolated from Chinese fetal kidney by the acid gmnidinium thiocyanate phenol-chloroform method. Two overlapping segments of human BMP- 1 cDNA were obtained by reverse transcription (RT)-PCR. Following application, the two segments were ligated to each other and subcloned into POEM-T easy vector to form PEGM-T easy/hBMP-7 recombinant plasmid. Sanger dideoxy chain-termination method was used to sequence the cDNA. Results. There was 750 bp fragment obtained RT-PCR using #2 primer from 5' end of BMP-7 gene (PCR by using # 2 and # 1) ,and 540 bp fragment from 3' end was generated by KT-PCR using # 4 primer (PCR using # 3 and # 4). Full-length cDNA encoding BMP-7 was obtained by religation of two segments. When compared with hBMP-7 sequence in Gene bank (XM30619) ,our full-length BMP-7 cDNA has a G instead of a T at nucleotide 862. This change results in valine substituting for phenylalanine in the protein. Conclusion. This is the first time that BMP-7 cDNA was successfully cloned from Chinese fetal kidney. BMP-7 cDNA plays an important role in healing injuries of the osteo-articular system. This makes BMP-7 is an attractive target far various clinical applications.

肾衰营养胶囊对肾性骨病模型大鼠肾功能和骨代谢的改善作用及其对BMP-7/Smads信号通路的影响

目的:探讨肾衰营养胶囊对肾性骨病模型大鼠的改善作用及其对骨形态发生蛋白(BMP)-7/Smads信号通路的影响,阐明肾性骨病模型大鼠肾功能和骨代谢与肾性骨病的关系。方法:选取50只SPF级SD大鼠,随机选取10只大鼠作为对照组,另外40只大鼠建立肾性骨病模型。将30只造模成功大鼠分为模型组、阳性对照组和肾衰营养胶囊组,每组各10只。对照组和模型组大鼠采用生理盐水灌胃,阳性对照组大鼠给予0.01 mg·kg^(-1)骨化三醇灌服,肾衰营养胶囊组大鼠给予1.2 g·kg^(-1)肾衰营养胶囊灌服,均灌胃12周。采用HE染色观察各组大鼠股骨组织病理形态表现,全自动生化分析仪和化学发光法检测各组大鼠血清中肾功能和钙磷代谢指标,实时荧光定量PCR(RT-qPCR)法检测各组大鼠股骨组织中BMP-7和Smad1/5/8 mRNA表达水平,Western blotting法检测各组大鼠股骨组织中BMP-7、磷酸化BMP-7(p-BMP-7)、Smad1/5/8和磷酸化Smad1/5/8(p-Smad1/5/8)蛋白表达水平。结果:HE染色,对照组大鼠骨小梁排列正常,成骨细胞和类骨质面积未发生改变;模型组大鼠骨小梁宽度和平均类骨质面积增加,成骨细胞数量减少;阳性对照组和肾衰营养胶囊组大鼠骨小梁宽度及平均类骨质面积减小,成骨细胞数量增加。与对照组比较,模型组大鼠血尿素氮(BUN)和血肌酐(Scr)水平均升高(P<0.05);与模型组比较,阳性对照组和肾衰营养胶囊组大鼠BUN及Scr水平均降低(P<0.05);与阳性对照组比较,肾衰营养胶囊组大鼠BUN和Scr水平均降低(P<0.05)。与对照组比较,模型组大鼠血清中钙离子(Ca2+)水平降低(P<0.05),磷离子(P3-)、甲状旁腺激素(PTH)和碱性磷酸酶(ALP)水平均升高(P<0.05);与模型组比较,阳性对照组和肾衰营养胶囊组大鼠血清中Ca2+水平升高(P<0.05),P3-、PTH和ALP水平均降低(P<0.05);与阳性对照组比较,肾衰营养胶囊组大鼠血清中Ca2+水平升高(P<0.05),P3-、PTH和ALP水平均降低(P<0.05)。RT-qPCR法,与对照组比较,模型组大鼠股骨组织中BMP-7和Smad1/5/8mRNA表达水平降低(P<0.05);与模型组比较,阳性对照药组和肾衰营养胶囊组大鼠股骨组织中BMP-7及Smad1/5/8 mRNA表达水平升高(P<0.05);与阳性对照组比较,肾衰营养胶囊组大鼠股骨组织中BMP-7和Smad1/5/8 mRNA表达水平升高(P<0.05)。Western blotting法,与对照组比较,模型组大鼠股骨组织中BMP-7、p-BMP-7、Smad1/5/8和p-Smad1/5/8蛋白表达水平均降低(P<0.05);与模型组比较,阳性对照组和肾衰营养胶囊组大鼠股骨组织中BMP-7、 p-BMP-7、 Smad1/5/8和p-Smad1/5/8蛋白表达水平均升高(P<0.05);与阳性对照组比较,肾衰营养胶囊组大鼠股骨组织中BMP-7、p-BMP-7、Smad1/5/8和p-Smad1/5/8蛋白表达水平均升高(P<0.05)。结论:肾衰营养胶囊可改善肾性骨病模型大鼠的肾功能和钙磷代谢紊乱,促进骨髓基质细胞增殖,改善骨代谢情况,其机制可能与激活BMP-7/Smads信号通路有关。

帕立骨化醇对肾性骨病大鼠骨代谢及TGF-β/BMP-7/Smad通路的影响

目的:探讨帕立骨化醇对肾性骨病(ROD)大鼠骨代谢及转化生长因子-β(TGF-β)/骨形态发生蛋白-7(BMP-7)/Smad通路的影响。方法:将90只大鼠随机分为6组:对照组、模型组、帕立骨化醇低剂量(0.2μg/kg)组、帕立骨化醇中剂量(0.4μg/kg)组、帕立骨化醇高剂量(0.8μg/kg)组、骨化三醇(10μg/kg)组,每组15只,对照组大鼠以普通饲料喂养,其余各组大鼠用含有腺嘌呤的饲料喂养,诱导建立ROD模型。分组进行药物治疗后,测定各组大鼠肾功能指标血尿素氮(BUN)与血肌酐(Scr)水平、血钙与血磷水平、股骨骨密度(BMD)、股骨生物力学指标最大载荷、弹性模量与屈服载荷、血清炎症因子IL-6、IL-17水平;采用蛋白免疫印迹法检测骨组织TGF-β/BMP-7/Smad通路蛋白表达情况。再次取45只大鼠随机分为3组:帕立骨化醇(0.8μg/kg)组、TGF-β抑制(LY2157299,150 mg/kg)组、帕立骨化醇(0.8μg/kg)+TGF-β抑制(LY2157299,150 mg/kg)组,每组15只,同样方法建立ROD模型。分组以药物治疗后,测定各组大鼠肾功能指标与股骨生物力学指标水平。结果:与对照组比较,模型组大鼠血钙水平、BMD、弹性模量、最大载荷、屈服载荷、骨组织TGF-β/BMP-7/Smad通路蛋白TGF-β及BMP-7表达、p-Smad3/Smad3显著降低(P<0.05),BUN与Scr水平、血磷水平、血清IL-6与IL-17水平显著升高(P<0.05)。与模型组比较,帕立骨化醇低、中、高剂量组和骨化三醇组大鼠血钙水平、BMD、弹性模量、最大载荷、屈服载荷、骨组织TGF-β/BMP-7/Smad通路蛋白TGF-β及BMP-7表达、p-Smad3/Smad3均升高,BUN与Scr水平、血磷水平、血清IL-6与IL-17水平均降低,且帕立骨化醇各组之间呈剂量依赖性(P<0.05),帕立骨化醇高剂量组和骨化三醇组比较,大鼠各指标差异无统计学意义(P>0.05)。与帕立骨化醇+TGF-β抑制组比较,帕立骨化醇组大鼠肾功能指标BUN、Scr与血磷水平降低(P<0.05),血钙水平、BMD、弹性模量、最大载荷、屈服载荷升高(P<0.05);TGF-β抑制组大鼠肾功能指标BUN、Scr与血磷水平升高(P<0.05),血钙水平、BMD、弹性模量、最大载荷、屈服载荷降低(P<0.05)。结论:帕立骨化醇可通过激活TGF-β/BMP-7/Smad信号通路抑制炎症反应,改善ROD大鼠肾功能及骨代谢异常,降低血磷水平,提高血钙水平及骨密度,修复骨生物力学,改善骨病症状。

血清CTRP3、BMP-7及PetCO_(2)对呼吸窘迫综合征患儿机械通气撤机的预测价值

目的:探讨新生儿呼吸窘迫综合征(NRDS)血清补体C1q-肿瘤坏死因子相关蛋白3(CTRP3)、骨形态发生蛋白-7(BMP-7)及呼吸末二氧化碳分压(PetCO_(2))对机械通气撤机的预测价值。方法:选取80例NRDS患儿为研究对象,依据撤机结局分为成功组54例、失败组26例。比较2组基线资料。分析不同组、不同严重程度患儿PetCO_(2)、血清CTRP3、BMP-7水平,分析其与生化指标、疾病严重程度相关性。评价PetCO_(2)、血清CTRP3、BMP-7对撤机结局的预测价值。采用SPSS24.0对数据进行分析,计数资料采用χ^(2)检验,计量资料采用t检验。结果:治疗12h、24h后成功组PetCO_(2)分别为(36.32±1.88)(mmHg)4、(33.06±1.79)(mmHg)4,血清BMP-7水平分别为(44.03±5.17)pg/mL、(41.16±4.88)pg/mL,血清℃TRP3水平分别为(345.59±60.03)μg/L、(377.41±75.80)μg/L;治疗12h、24h后失败组PetCO_(2),分别为(38.62±1.93)(mmHg)4、(38.57±1.90)(mmHg)4,血清BMP-7水平分别为(48.82±5.96)pg/mL、(48.39±5.63)pg/mL,血清CTRP3水平分别为(316.68±55.56)μg/L、(320.03±66.67)μg/L,治疗12h、24h后成功组PetCO_(2)、血清BMP-7水平低于失败组,血清CTRP3水平高于失败组(P<0.05);PetCO_(2)、BMP-7与PaO_(2)呈负相关,且与PaCO_(2)、SNAPPE-ⅡI评分、TGF-β1、IL-6、TNF-α、病情严重程度呈正相关,而CTRP3与之相反(P<0.05);治疗12h、24h后血清CTRP3、BMP-7及PetCO_(2),联合预测撤机结局的AUC大于单项预测(P<0.05)。结论:血清CTRP3、BMP-7及PetCO_(2),对NRDS患儿撤机结局预测具有应用价值。

早产儿BPD的危险因素及血浆BMP-7 miR-15b 25-(OH)D_(3)对其诊断价值分析

目的 探讨早产儿支气管肺发育不良(BPD)的危险因素及血浆骨形成蛋白-7(BMP-7)、微小核糖核酸-15b(miR-15b)、25羟基维生素D_(3)[25-(OH)D_(3)]对其诊断价值。方法 回顾性分析2022年1月至2023年3月河北省沧州中西医结合医院收治的136例早产儿的临床资料,根据其是否伴有BPD,将其分为BPD组(n=45)和非BPD组(n=91)。分析早产儿BPD的单因素,采用多因素logistic回归分析早产儿BPD的危险因素,绘制受试者工作特征(ROC)曲线分析血浆BMP-7、miR-15b、25-(OH)D_(3)对早产儿BPD的诊断价值。结果 两组患儿机械通气时间、吸氧时间、有无脓毒血症及血浆BMP-7、miR-15b、25-(OH)D_(3)水平比较,差异有统计学意义(P<0.05)。多因素logistic回归分析结果显示,机械通气时间长、血浆BMP-7、miR-15b水平偏高、血浆25-(OH)D_(3)水平偏低是早产儿BPD的独立危险因素(OR=2.625、2.208、2.280、2.517,P<0.05)。血浆BMP-7、miR-15b、25-(OH)D_(3)联合诊断早产儿BPD的曲线下面积为0.947,高于血浆BMP-7、miR-15b、25-(OH)D_(3)单独检测(0.822、0.849、0.824,P<0.05)。结论 早产儿BPD的危险因素与机械通气时间长、血浆BMP-7、miR-15b水平偏高、血浆25-(OH)D_(3)水平偏低有关,且血浆BMP-7、miR-15b、25-(OH)D_(3)联合检测对早产儿BPD的诊断价值更高。

血清TXNIP、sTREM-1、BMP-7水平变化与原发性肾病综合征患者疾病转归的关系

目的 探讨血清硫氧还蛋白相互作用蛋白(TXNIP)、可溶性髓系细胞触发受体-1(sTREM-1)、骨形态发生蛋白-7(BMP-7)水平变化与原发性肾病综合征(PNS)患者疾病转归的相关性。方法 选取医院2019年4月至2022年4月收治的91例PNS患者为研究组,同期91例健康体检者为对照组,比较两组入院时血清TXNIP、sTREM-1、BMP-7、肾功能指标[尿素氮(BUN)、血肌酐(Scr)、尿酸(UA)]水平,分析血清各指标水平与肾功能的相关性;比较复发和未复发患者(入院时、治疗3个月、6个月后)血清各指标水平;分析治疗后血清各指标水平联合预测PNS患者复发的价值,分析各指标不同水平患者复发危险度。结果 入院时研究组血清TXNIP、sTREM-1高于对照组,血清BMP-7低于对照组(P<0.05);入院时研究组血清BUN、Scr、UA高于对照组(P<0.05);入院时血清TXNIP、sTREM-1水平与BUN、Scr、UA水平均呈正相关,血清BMP-7水平与BUN、Scr、UA水平均呈负相关(P<0.05);入院时、治疗3个月、6个月后复发患者血清TXNIP、sTREM-1水平高于未复发患者,BMP-7水平低于未复发患者(P<0.05);治疗3、6个月后血清各指标联合预测PNS患者复发价值大于单独预测;治疗3、6个月后血清各指标高水平患者复发危险度是低水平的1.906、2.114、0.581倍/2.571、3.222、0.480倍。结论 血清TXNIP、sTREM-1、BMP-7水平与PNS患者病情程度密切相关,可作为评估PNS病情转归的有效指标。

新生儿呼吸窘迫综合征患儿血清HMGB1、RAGE、BMP-7表达及与肺部超声评分的相关性

目的 分析新生儿呼吸窘迫综合征(NRDS)患儿血清高迁移率族蛋白1(HMGB1)、晚期糖基化终末产物受体(RAGE)、骨形态发生蛋白-7(BMP-7)表达及与肺部超声评分的相关性。方法 选择2018年6月至2023年6月收治的100例NRDS患儿纳入观察组,同期选择100例健康新生儿纳入对照组,均进行血清HMGB1、RAGE、BMP-7检测和肺部超声检查及评分。根据NRDS患儿肺部超声评分将其分为低评分组(评分<12分)、中评分组(评分12~24分)及高评分组(评分>24分)。比较两组研究对象的血清HMGB1、RAGE、BMP-7水平和肺部超声评分;比较不同肺部超声评分患儿的血清HMGB1、RAGE、BMP-7水平;分析观察组中血清HMGB1、RAGE、BMP-7表达与肺部超声评分的相关性。结果 观察组的血清HMGB1、RAGE及BMP-7水平及肺部超声评分高于对照组(P<0.05)。肺部超声显示低评分组47例,中评分组33例,高评分组20例;低评分组的血清HMGB1、RAGE及BMP-7水平低于中评分组和高评分组(P<0.05);中评分组的血清HMGB1、RAGE及BMP-7水平低于高评分组(P<0.05)。Spearman相关性分析结果显示,血清HMGB1、RAGE、BMP-7表达与肺部超声评分呈正相关(P<0.05)。结论 NRDS患儿血清HMGB1、RAGE、BMP-7表达及肺部超声评分均会明显增高,血清HMGB1、RAGE、BMP-7表达与肺部超声评分呈正相关。

健脾化湿法治疗慢性肾衰竭患者的疗效及对血清TGF-β1、BMP-7的水平的影响

目的 探讨健脾化湿法治疗慢性肾衰竭(CRF)患者的疗效及对血清转化生长因子-β1(TGF-β1)、骨形成蛋白-7(BMP-7)的水平的影响。方法 选择2020年10月—2021年9月在该院就诊的82例CRF患者为研究对象,采用奇偶数法将其随机分为两组,即观察组和对照组各41例。对照组予以西药常规治疗,观察组在对照组基础上采用健脾化湿益肾泄浊方治疗,均治疗3个月。比较两组治疗后的疗效,治疗前后的主要症状中医证候积分,血清TGF-β1、BMP-7、纤维连接蛋白(FN)、肌酐(SCr)、血尿素氮(BUN)和白蛋白水平。结果 治疗后,观察组的治疗总有效率高于对照组(P<0.05);同时,观察组患者的倦怠乏力等各症状中医证候积分均低于对照组(P<0.05);观察组血清TGF-β1、Scr、BUN、FN和24 h尿蛋白定量水平均低于对照组(P<0.05),BMP-7、白蛋白水平高于对照组(P<0.05)。结论 采用健脾化湿法治疗CRF可明显改善患者肾功能和中医证候,降低血清TGF-β1的表达水平,提升血清BMP-7水平,临床疗效显著。

BMP7-Ids信号通路对子宫内膜癌细胞增殖、侵袭的作用

目的探讨骨成型蛋白7(BMP7)-Ids信号通路对子宫内膜癌细胞增殖、侵袭的作用。方法以人子宫内膜癌细胞Ishikawa作为研究对象,将细胞分为四组,即空白对照组(不加BMP7)、A组(2 ng/ml BMP7)、B组(5 ng/ml BMP7)、C组(10ng/mlBMP7)。采用qRT-PCR法检测分化抑制因子(Id)1、Id3的mRNA;Western blot法检测Id1、Id3蛋白表达情况;划痕实验检测细胞迁移能力;采用MTT法检测细胞增殖;采用Transwell实验观察细胞的侵袭情况。每组试验均重复4次,取平均值。结果A组、B组和C组Id1、Id3的mRNA和蛋白表达水平均高于空白对照组,其中C组指标高于A组、B组,B组指标高于A组,差异均有统计学意义(P<0.05);24、36和48h,A、B和C组细胞增殖率高于空白对照组,C组细胞增殖率高于A组和B组,且B组细胞增殖率高于A组,差异有统计学意义(P<0.05)。A、B和C组细胞侵袭细胞数和划痕相对愈合面积高于空白对照组,差异有统计学意义(P<0.05);C组侵袭细胞数和划痕相对愈合面积高于A组和B组,且B组侵袭细胞数和划痕相对愈合面积高于A组,差异有统计学意义(P<0.05)。结论子宫内膜癌细胞中BMP7-Ids信号通路的表达上调可激活细胞的增殖和侵袭作用,从而促进子宫内膜癌病情的发展。

维持性腹膜透析患者血清Irisin和BMP-7水平与血管钙化相关性分析

目的:分析维持性腹膜透析(MPD)患者中鸢尾素(Irisin)和骨形态发生蛋白7(BMP-7)水平与血管钙化相关性。方法:选取2019年6月—2021年12月我院腹膜透析中心收治的透析时间大于3个月患者128例,检测所有患者血钙(Ca)、血磷(P)、血红蛋白(Hb)、白蛋白(Alb)、总胆固醇(Tch)、甘油三酯(TG)等指标。酶联免疫吸附试验(ELISA)检测血清Irisin、BMP-7水平,腹主动脉钙化通过侧位腹平片X线评价。结果:根据AACS情况将128例MPD患者分为非钙化组(AACS=0分)50例和非钙化组(AACS≥1分)78例。在原发病方面,钙化组糖尿病肾病患者23例,非钙化组糖尿病肾病患者5例,两组存在显著差异(P<0.05);在性别方面,钙化组男性51例,非钙化组男性23例,两组存在差异(P<0.05);在年龄、透析时间方面,钙化组年龄>65岁患者例数和透析时间显著多于非钙化组(P<0.05)。钙化组患者血清中Ca、P、全段甲状旁腺激素(iPTH)显著高于非钙化组(P<0.05);钙化组患者血清中残肾Irisin、BMP-7、Alb显著低于非钙化组(P<0.05)。Pearson相关分析显示,Irisin和BMP-7水平与年龄、透析时间、血钙、血磷、钙磷乘积、iPTH呈负相关(P<0.05)。结论:MPD患者血管钙化的危险因素有高龄、长透析龄、糖尿病肾病、P、iPTH。Irisin、BMP-7的水平与血管钙化、钙磷代谢及甲状旁腺激素水平呈负相关,Irisin、BMP-7水平低更易发生血管钙化。