Effect of vitamin D3 on production of progesterone in porcine granulosa cells by regulation of steroidogenic enzymes

1,25-dihydroxyvitamin D3 （VD3）, an active form of Vitamin D, is photosynthesized in the skin of vertebrates in response to solar ultraviolet B radiation （UV-B）. VD3 deficiency can cause health problems such as immune disease, metabolic disease, and bone disorders. It has also been demonstrated that VD3 is involved in reproductive functions. Female sex hormones such as estrogen and progesterone are biosynthesized mainly in ovarian granulosa cells as the ovarian follicle develops. The functions of sex hormones include regulation of the estrus cycle and puberty as well as maintenance of pregnancy in females. In this study, we isolated granulosa cells from porcine ovaries and cultured them for experiments. To examine the effects of VD3 on ovarian granulosa cells, the mRNA and protein levels of genes were analyzed by Real-time PCR and Western blotting assay. Production of progesterone from granulosa cells was also measured by ELISA assay. As a result, transcriptional and translational regulation of progesterone biosynthesis-related genes in granulosa cells was significantly altered by VD3. Furthermore, progesterone concen- trations in porcine granulosa cell-cultured media decreased in response to VD3. These results show that VD3 was a strong regulator of sex steroid hormone production in porcine granulosa cells, suggesting that vitamin D deficiency may result in inappropriate sexual development of industrial animals and eventually economic loss.

Vitamin D3 supplementation influences ovarian histomorphometry and follicular development in prepubertal albino rats

Objective:To evaluate the development of ovarian follicles in female albino rats following vitamin D3 supplementation.Methods:Eighteen prepubertal female albino rats,aged 3-4 weeks,weighing(70.25±9.16)g,were assigned to three groups(n=6 in each group).Group A was treated with 5.00 mL/kg of distilled water and served as the control group,group B was treated with 0.025 mg/kg of vitamin D3 dissolved in distilled water,and group C was treated with 0.125 mg/kg of vitamin D3 dissolved in distilled water.All treatments were administered orally,twice weekly for 28 days.Blood and ovaries were harvested under anaesthesia.Serum vitamin D3 levels were determined by using spectrophotometric method.Ovaries were processed for histology and every10th hematoxylin and eosin stained-section was selected for histomorphometry.The number of follicles at each developmental stage was estimated.Results:Both 0.025 mg/kg and 0.125 mg/kg of vitamin D3 significantly increased serum concentrations of vitamin D3 and calcium(P<0.05),but did not alter inorganic phosphorus concentration(P>0.05).The control group had fewer growing follicles(primary,secondary and antral follicles)and more non-growing follicles(primordial and atretic follicles)when compared with the vitamin D3-supplemented groups(P<0.05).Vitamin D3 at 0.025 mg/kg significantly increased antral follicles and corpora lutea counts(P<0.05).Vitamin D3 at 0.125 mg/kg significantly increased total,primordial and atretic follicles counts(P<0.05),but significantly decreased primary,secondary,antral follicles count,ovarian weight,relative ovarian weight,and ovarian surface area when compared with the control group and rats treated with 0.025 mg/kg of vitamin D3(P<0.05).Conclusions:Vitamin D3 supplementation at 0.025 mg/kg can enhance optimal ovarian follicle recruitment and development in female rats.

Enhanced Response of Acute Monocytic Leukemia Cells to Low-dose Cytarabine by 1,25-dihydroxyvitamin D3

Low-dose cytarabine combined with differentiating or DNA hypomethylating agents,such as vitamin D compounds,is a potential regimen to treat acute myeloid leukemia（AML） patients who are unfit for high-intensity chemotherapy.The present study aimed to determine which subset of AML would be most responsive to low-dose cytarabine with the differentiating agent 1,25-dihydroxyvitamin D3（1,25-D3）.Here,firstly,c Bio Portal database was used and we found out that vitamin D receptor（VDR） was highly expressed in acute monocytic leukemia（M5） and high VDR expression was associated with a poor survival of AML patients.Then,we confirmed that 1,25-D3 at clinical available concentration could induce more significant differentiation in acute monocytic leukemia cell lines（U937,MOLM-13,THP-1） and blasts from M5 patients than in non-monocytic cell lines（KG1 a and K562） and blasts from M2 patient.Finally,it was shown that the combination of 1,25-D3 and low-dose cytarabine further increased the differentiating rate,growth inhibition and G0/G1 arrest,while mild changes were found in the apoptosis in acute monocytic leukemia cell lines.Our study demonstrates that the enhanced response of acute monocytic leukemia cells to low-dose cytarabine by 1,25-D3 might indicate a novel therapeutic direction for patients with acute monocytic leukemia,especially for elderly and frail ones.

A Syndrome of Vitamin D3 Deficiency/Fibromyalgia/Hyperparathyroidism Mimicking Rheumatoid Arthritis, a Clinical Prospective Study

Objectives: Rheumatoid arthritis is sometimes misdiagnosed for other diseases, like psoriatic arthritis, erosive OA, viral arthritis, reactive arthritis, IBD arthritis, Lyme’s disease, and palindromic rheumatism. Secondary hyperparathyroidism was not included in the differential diagnosis of RA, though it sometimes presents with joint pains and tenderness, and even arthritis. Fibromyalgia is a psychosomatic disorder characterized by chronic widespread pain and tender areas. Mimicry of some manifestations of these diseases sometimes results in misdiagnosis as RA. Methods: Two hundred patients previously diagnosed as RA from outpatient clinics in Al-Azhar Faculty of Medicine, attended as not responding to medical treatment of RA. All patients were subjected to a re-evaluation of disease activity including HAQ, and DAS 28, CDAI, and SDAI. Also, we measured serum vitamin D3, PTH, total and ionized Calcium, Phosphorus, Uric acid, ACPA, and other routine lab. All patients were exposed to various radiological imaging modalities. Results: Cases not responding to RA treatment were reevaluated and were found to have a syndrome of fibromyalgia associated with vitamin D3 deficiency and secondary hyperparathyroidism. Conclusions: Fibromyalgia/Hyperparathyroidism syndrome is an underdiagnosed disease, which results from chronic vitamin D3 deficiency. SHPT can cause bone erosions, which are mostly shaggy in appearance and distributed in the radiocarpal, radioulnar, metacarpophalangeal and distal interphalangeal joints, in contrast to that which predominate proximal IP joints of rheumatoid arthritis. Radiology of FM/HPT syndrome patients revealed a sign of spur-like excrescences in terminal finger tufts unilaterally or bilaterally, which we think is pathognomonic.

The Role of Vitamin D3 Therapy in Pediatric Bronchiectasis Severity (CF versus non-CF Patients)

Objective: To determine and compare the effect of vitamin D3 supplementation on modifying the disease severity in cystic fibrosis (CF) and non-CF bronchiectasis pediatric patients. Methods: A randomized clinical trial evaluating the role of oral vitamin D3 supplementation for six months, was performed in forty patients with CF and non-CF bronchiectasis under the age of 18 years with vitamin D deficiency or insufficiency. The primary outcome was to reach the sufficient Vitamin D level, the secondary outcome was to reevaluate bronchiectasis severity by following up the frequency, severity of pulmonary exacerbations and lung function after vitamin D3 supplementation. Results: Forty patients completed the trial. The percentage of improvement of vitamin D level after vitamin D3 supplementation for six months was significantly higher in CF (88.3%) than non-CF bronchiectasis patients (59.82%) (P = 0.03). Additionally, moderate to severe pulmonary exacerbations significantly decreased by more than 60%, 45% (P = 0.001, 0.005) and frequent exacerbations decreased by 15%, 10% (P = 0.327, 0.490), while the forced expiratory volume in 1 (FEV1) significantly increased by 17% and 15% in non CF bronchiectasis and CF patients respectively (p < 0.001). Conclusions: Vitamin D3 therapy was effective in decreasing the frequency and severity of pulmonary exacerbations and preserving lung function. Thereby, improving the disease severity even more in non-CF bronchiectasis than CF patients.

CYP24A1 inhibition facilitates the anti-tumor effect of vitamin D3 on colorectal cancer cells

AIM:The effects of vitamin D3 have been investigated on various tumors, including colorectal cancer (CRC). 25-hydroxyvitamin-D3-24-hydroxylase (CYP24A1), the enzyme that inactivates the active vitamin D3 metabolite 1,25-dihydroxyvitamin D3 (1,25-D3), is considered to be the main enzyme determining the biological halflife of 1,25-D3. During colorectal carcinogenesis, the expression and concentration of CYP24A1 increases significantly, suggesting that this phenomenon could be responsible for the proposed efficacy of 1,25-D3 in the treatment of CRC. The aim of this study was to investigate the anti-tumor effects of vitamin D3 on the human CRC cell line Caco-2 after inhibition of the cytochrome P450 component of CYP24A1 activity. METHODS:We examined the expression of CYP24A1 mRNA and the effects of 1,25-D3 on the cell line Caco-2 after inhibition of CYP24A1. Cell viability and proliferation were determined by means of sulforhodamine-B staining and bromodeoxyuridine incorporation, respectively, while cytotoxicity was estimated via the lactate dehydrogenase content of the cell culture supernatant. CYP24A1 expression was measured by realtime reverse transcription polymerase chain reaction. A number of tetralone compounds were synthesized to investigate their CP24A1 inhibitory activity. RESULTS:In response to 1,25-D3, CYP24A1 mRNA expression was enhanced significantly, in a time- and dose-dependent manner. Caco-2 cell viability and proliferation were not influenced by the administration of 1,25-D3 alone, but were markedly reduced by coadministration of 1,25-D3 and KD-35, a CYP24A1-inhibiting tetralone. Our data suggest that the mechanism of action of co-administered KD-35 and 1,25-D3 does not involve a direct cytotoxic effect, but rather the inhibition of cell proliferation. CONCLUSION:These findings demonstrate that the selective inhibition of CYP24A1 by compounds such as KD-35 may be a new approach for enhancement of the anti-tumor effect of 1,25-D3 on CRC.

Preparation and Spectroscopic of Vanadyl（Ⅱ） Vitamin D3 Amino Acid Mixed Complexes as Insulin Mimetic Drug

A new six intraperitoneal injection insulin-mimetic vanadyl(Ⅱ) compounds [(VD3^-1)(VO^+2)(AAn^-1)](where(n=1~6);AA1=isoleucine, AA2=threonine, AA3=proline, AA4=phenylalanine, AA5=lysine and AA6=glutamine) were synthesized by the chemical reactions between vitamin D3(VD3), VOSO4 and amino acids(AAn) with equal molar ratio 1∶1∶1 in neutralized media. The structures of these complexes were elucidated by spectroscopic methods like, infrared and solid reflectance spectroscopes. Magnetic moments and electronic spectra reveal square pyramid geometrical structure of the complexes. The infrared spectra assignments of these complexes revealed that the chelation towards vanadyl(Ⅳ) ions existed via deprotonation of the hydroxyl group of VD3 drug ligand and so amino acids act as bidentate ligand via N-amino and O-carboxylate groups. The anti-diabetic efficiency of these complexes were evaluated against streptozotocin induced diabetic male albino rats.

维生素D3减轻高糖暴露诱导氧化应激促进人脐带间充质干细胞的成骨分化

背景:糖尿病骨质疏松症逐渐受到公众的关注,然而鲜有研究报道高糖环境对人脐带间充质干细胞成骨分化的影响和相应的治疗策略。目的:探讨维生素D3恢复高糖环境中人脐带间充质干细胞成骨分化的潜力。方法:通过CCK-8法检测人脐带间充质干细胞活力来筛选适宜的维生素D3干预浓度。在高葡萄糖条件下,通过RT-qPCR、蛋白质印迹、免疫荧光、JC-1线粒体膜电位、茜素红染色和β-半乳糖苷酶染色等实验评估维生素D3干预后人脐带间充质干细胞的成骨分化潜能、细胞内活性氧积累、线粒体膜电位改变和细胞衰老情况,并探讨了潜在的机制。结果与结论:①维生素D3在0.1μmol/L至1 mmol/L范围内均能显著促进人脐带间充质干细胞的增殖;②高糖环境下调了人脐带间充质干细胞中成骨相关基因α1-Ⅰ型胶原蛋白、碱性磷酸酶、Runt相关转录因子2、骨钙蛋白的mRNA和蛋白表达水平,诱发了氧化应激和细胞衰老;③维生素D3在10μmol/L的干预浓度下显著恢复了高糖条件下人脐带间充质干细胞的成骨表型,并通过激活Nrf2/HO-1信号通路减轻了细胞内的氧化应激和细胞衰老;④结果表明,高糖环境中人脐带间充质干细胞的成骨分化能力降低,维生素D3可以部分提高其成骨分化能力并减轻细胞损伤。

VD3调控ROS介导的TXNIP/NLRP3通路抑制高糖诱导肾系膜细胞纤维化的机制研究

目的:探究活性维生素D3(VD3)对高糖诱导下肾系膜细胞纤维化特征的影响,并探讨相关作用机制。方法:培养大鼠肾小球系膜细胞系HBZY-1,将其分为正常葡萄糖培养(NG)组、正常葡萄糖联合VD3培养(NG+VD3)组、高葡萄糖培养(HG)组、高葡萄糖联合VD3培养(HG+VD3)组、高葡萄糖联合N-乙酰半胱氨酸培养(HG+NAC)组。CCK-8法检测各组细胞的增殖活性,流式细胞术检测各组细胞的凋亡率,DCFH-DA法检测各组细胞中活性氧(ROS)含量,酶联免疫吸附法(ELISA)检测各组细胞上清液中白细胞介素-1β(IL-1β)、IL-18、肿瘤坏死因子-α(TNF-α)水平,实时荧光定量聚合酶反应(RT-qPCR)检测各组细胞中纤维化标志物转化生长因子-β1(TGF-β1)、纤维连接蛋白(FN)、细胞间粘附分子-1(ICAM-1)及硫氧还蛋白互作蛋白(TXNIP)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)的mRNA表达,蛋白质免疫印记(Western blot)检测各组细胞中TGF-β1、FN、ICAM-1及TXNIP、NLRP3的蛋白表达。结果:与NG组比较,HG组增殖活性显著升高(P<0.05),细胞凋亡率显著减少(P<0.05),ROS含量显著上升(P<0.05),细胞上清液中IL-1β、IL-18、TNF-α水平显著升高(P<0.05),细胞中TGF-β1、FN、ICAM-1、TXNIP、NLRP3的mRNA和蛋白相对表达水平显著上调(P<0.05);与HG组比较,HG+VD3组和HG+NAC组增殖活性显著降低(P<0.05),细胞凋亡率显著增加(P<0.05),ROS含量显著下降(P<0.05),细胞上清液中IL-1β、IL-18、TNF-α水平显著降低(P<0.05),同时,细胞中TGF-β1、FN、ICAM-1及TXNIP、NLRP3的mRNA和蛋白相对表达水平显著下调(P<0.05)。结论:VD3能够减轻高糖诱导的肾系膜细胞过度增殖及纤维化,该机制可能与调控ROS/TXNIP/NLRP3通路有关。

维生素D3通过增强肝组织维生素D受体活性阻断JAK/STAT3通路减轻高胆固醇血症小鼠幽门螺杆菌感染相关胃炎

目的 探讨维生素D3(VD3)能否通过降低血脂抑制Janus激酶/信号转导子与转录激活子3(JAK/STAT3)信号通路从而减轻幽门螺杆菌(Hp)感染。方法 建立高胆固醇小鼠模型和Hp感染小鼠模型,采用VD3灌胃8周。采用实时定量PCR检测小鼠肝组织维生素D受体(VDR)、胰岛素诱导基因2(Insig-2)及小鼠胃组织胃泌素的mRNA表达,Western blot法检测胃组织JAK、STAT3、环加氧酶2(COX2)蛋白的表达,生化分析法检测小鼠血清胆固醇水平,ELISA检测各组小鼠血清白细胞介素6(IL-6)及IL-8水平,HE染色小鼠肝组织及胃组织的病变情况。结果 高胆固醇组及高胆固醇联合Hp感染组小鼠在灌胃VD3后,小鼠肝组织VDR、Insig-2的水平明显上升,胃泌素水平表达降低;胃组织JAK、STAT3及COX2蛋白的表达降低,血清中胆固醇水平降低,IL-6水平无明显变化,IL-8水平降低;与对照组相比,高胆固醇联合Hp感染组肝细胞气球样变减少,胃组织炎症减轻,胆固醇组、Hp感染组胃组织炎症也减轻。结论 VD3通过增强肝组织VDR的活性,阻断JAK/STAT3信号通路,抑制炎症因子表达减轻胃炎的程度。

1,25-Dihydroxyvitamin D3 protects obese rats from metabolic syndrome via promoting regulatory T cell-mediated resolution of inflammation

Vitamin D_3 has been found to produce therapeutic effects on obesity-associated insulin resistance and dyslipidemia through its potent anti-inflammatory activity, but the precise immunomodulatory mechanism remains poorly understood. In the present study we found that 1,25-dihydroxyvitamin D_3[1,25(OH)_2D_3], the biologically active form of vitamin D_3, significantly attenuated monosodium glutamate(MSG)-induced obesity and insulin resistance as indicated by body weight reduction, oral glucose tolerance improvement, and a glucose infusion rate increase as detected with hyperinsulinemiceuglycemic clamp. Moreover, 1,25(OH)_2D_3 not only restored pancreatic islet functions but also improved lipid metabolism in insulin-targeted tissues. The protective effects of 1,25(OH)_2D_3 on glycolipid metabolism were attributed to its ability to inhibit an obesity-activated inflammatory response in insulin secretory and targeted tissues, as indicated by reduced infiltration of macrophages in pancreas islets and adipose tissue while enhancing the expression of Tgf-β1 in liver tissue, which was accompanied byincreased infiltration of Treg cells in immune organs such as spleen and lymph node as well as in insulintargeted tissues such as liver, adipose, and muscle. Together, our findings suggest that 1,25(OH)_2D_3 serves as a beneficial immunomodulator for the prevention and treatment of obesity or metabolic syndrome through its anti-inflammatory effects.

B超引导下甲状旁腺内注射维生素D3对维生素D缺乏相关甲状旁腺功能亢进症患者钙磷代谢及肌肉功能的影响

目的:探讨B超引导下甲状旁腺内注射1,25双羟维生素D3[1,25(OH)_(2)D_(3)]对维生素D缺乏相关甲状旁腺功能亢进症患者钙磷代谢、肌肉功能的影响。方法:选取2020年1月至2022年6月于联勤保障部队第904医院肾脏内分泌科确诊为维生素D缺乏继发甲状旁腺功能亢进症的60例患者,按随机数表法将其分为观察组和对照组,每组30例。观察组患者给予B超引导下甲状旁腺内注射1,25(OH)_(2)D_(3)治疗,对照组患者给予口服1,25(OH)_(2)D_(3)治疗,于治疗前、治疗后2周观察两组患者超声影像图特征,检测钙磷水平、甲状旁腺素(PTH)、碱性磷酸酶(ALP),25羟维生素D(25OHD)、1,25(OH)_(2)D_(3)水平,以及24 h尿钙、尿磷水平,测量两组患者的骨骼肌力量和功能并进行对比分析。结果:两组患者治疗前,超声图像显示甲状腺左叶下极背侧低回声病灶,动脉期呈均匀高增强,治疗后,病灶动脉期无明显增强,呈“空洞”表现;观察组和对照组治疗后血钙水平分别为(2.06±0.11)mmol/L、(2.21±0.12)mmol/L,高于治疗前(1.92±0.02)mmol/L、(1.93±0.03)mmol/L,血磷水平分别为(0.92±0.11)mmol/L、(1.11±0.07)mmol/L,低于治疗前(1.24±0.02)mmol/L、(1.25±0.03)mmol/L,差异均有统计学意义(t=6.859、12.399、15.677、10.069,P<0.05),且观察组治疗后血钙、血磷水平均低于对照组,差异均有统计学意义(t=5.047、7.982,P<0.05);两组治疗后血PTH、ALP水平均低于治疗前,且观察组治疗后PTH、ALP水平均低于对照组,差异有统计学意义(t=28.125、12.605,P<0.05);两组治疗后血25OHD、1,25(OH)_(2)D_(3)及24 h的UCa水平均高于治疗前,24 h的UP水平均低于治疗前,且观察组治疗后血25OHD、1,25(OH)_(2)D_(3)及24 h的UCa高于对照组,24 h的UP水平低于对照组,差异有统计学意义(t=9.6763、5.269、45.086、3.387,P<0.05);两组治疗后L1~4、股骨颈骨密度值均高于治疗前,且观察组治疗后腰椎L1~4、股骨颈骨密度值均高于对照组,差异有统计学意义(t=26.944、2.355,P<0.05);两组治疗后骨骼肌握力、步速均高于治疗前,且观察组治疗后骨骼肌握力、步速均高于对照组,差异有统计学意义(t=2.711、3.093,P<0.05)。结论:维生素D缺乏相关甲状旁腺功能亢进症患者腺体出现钙化,超声表现为不同形状强回声。B超引导下甲状旁腺内注射1,25(OH)_(2)D_(3)可减少骨折的发生。

阿法骨化醇软胶囊联合阿仑膦酸钠维D3片治疗老年骨质疏松症的临床效果分析

目的探讨老年骨质疏松症(Osteoporosis,OP)采用阿法骨化醇软胶囊联合阿仑膦酸钠维D3片治疗的临床效果。方法选取2021年4月—2023年4月北京积水潭医院贵州医院收治的116例老年OP患者为研究对象,以随机数表法分成研究组(n=58,阿法骨化醇软胶囊联合阿仑膦酸钠维D3片治疗)和对照组(n=58,阿仑膦酸钠维D3片治疗),比较两组临床疗效、骨密度、视觉模拟评分法(Visual Analogue Scales,VAS)、Oswestry功能障碍指数(Oswestry Disability Index,ODI)评分。结果研究组治疗总有效率为96.55%(56/58),较对照组的84.48%(49/58)更高,差异有统计学意义(χ^(2)=4.921,P<0.05)。治疗后,两组骨密度增加,且研究组较对照组更高,差异有统计学意义(P均<0.05);治疗后两组VAS、ODI评分下降,且研究组低于对照组,差异有统计学意义(P均<0.05)。结论老年OP患者采用阿法骨化醇软胶囊与阿仑膦酸钠维D3片联合治疗,能够促进临床疗效提高,改善其骨密度水平,降低疼痛程度,提高活动能力。

Efficacy and safety of vitamin D3 in patients with diabetic nephropathy： a meta-analysis of randomized controlled trials

Background Several studies found that vitamin D3 might alter glucose metabolism,protect kidney from injury and even proposed the mechanisms.But results from previous studies have been conflicting.The aim of this study was to evaluate the efficacy and safety of vitamin D3 in patients with diabetic nephropathy.The underlying mechanism of vitamin D3 decreasing proteinuria is also discussed.Methods We conducted a search of English and Chinese articles using database of Pubmed,Embase,Sinomed,CNKI,Wanfang and clinical trial register centers,for randomized controlled trials of vitamin D3 in diabetic nephropathy patients.Two reviewers performed independently.Meta-analysis was used when studies were homogeneous enough.Results Twenty studies,including 1 497 patients with diabetic nephropathy,were involved in this systemic review.Vitamin D3-treated patients with diabetic nephropathy had a statistically significant reduction in 24-hour proteinuria （weighted mean difference-0.44,95% CI-0.54 to-0.34,Z=8.80,P 〈0.000 01） and urine albumin/creatine ratio （standardized mean difference-0.29,95% CI-0.48 to-0.10,Z=2.96,P=0.003）.But vitamin D3 supplementation did not significantly reduce blood pressure and hemoglobin A1c compared with control group.The potential mechanisms about the renal protection of vitamin D3,including the inhibition of rennin-angiotensin system,the protection of kidney from inflammation,fibrosis and the structure change of kidney are discussed.In addition,vitamin D3 did not significantly increase the incidence of adverse effects,including total adverse effects,gastrointestinal adverse effects and fluctuation of blood pressure.Conclusions Vitamin D3 can ameliorate proteinuria and protect kidney from injury in patients with diabetic nephropathy.This renoprotective effect is independent of blood pressure and glucose reduction.And it does not increase any adverse effects than control,even in combination therapy with angiotensin converting enzyme inhibitors/angiotensin receptor blockers.But due to the limited randomized controlled trials of high quality,more clinical researches should be taken in the future.

1,25-Dihydroxyvitamin D3 pretreatment enhances the efficacy of allergen immunotherapy in a mouse allergic asthma model

Background Allergen-specific immunotherapy can induce immune tolerance to specific allergens by regulating immune status of individuals. However, its clinical application is limited due to individual differences in efficacy among patients and un-confirmed safety. 1,25 Dihydroxyvitamin D3 （1,25（OH）2D3） has been shown to be involved in a variety of physiological processes, including immune response regulation. In the present study we explored the role of 1,25（OH）2D3 pretreatment for immunotherapy.Methods Seventy-five BALB/c mice were randomly divided into five groups （15 mice per group）. The mouse allergic asthma model was established by intra-peritoneal injection of ovalbumin （OVA, 10 μg） and aluminium hydroxide （2 mg）as an adjuvant. Intra-peritoneal injection of 50 ng of 1,25（OH）2D3 served as a pretreatment, subcutaneous injection of OVA （100 μg） as an immunotherapy, and 1% OVA inhalation as a challenge. Histopathological analysis was performed on four mice per group. The number of cells and their classification in bronchoalvolar lavage （BAL） fluid were assayed.Levels of serum OVA-specific immunoglobulin E （slgE） and IFN-Y, IL-4, IL-5 and IL-10 in BAL fluid were measured by ELISA.Results After 1,25（OH）2D3 pretreatment, immunotherapy could significantly inhibit the infiltration of inflammatory cells into lung tissues and BAL fluid of mice with allergic asthma when compared with un-treated animals （eosinophils：（7.46±1.34）×104/ml vs. （13.41±1.67）×104/ml, P ＜0.05）. In addition, levels of IL-4 （（36.g1±7.87） pg/ml vs. （43.70±6.42）pg/ml, P ＞0.05） and IL-5 （（41.97±7.93） pg/ml vs. （60.14±8.35） pg/ml, P ＜0.05） in BAL fluid and serum slgE （（0.42±0.05）vs. （0.75±0.06） OD units, P ＜0.05） were profoundly reduced. However, the IL-10 level in BAL fluid was significantly increased （（67.74±6.57） pg/ml vs. （44.62±8.81） pg/ml, P ＜0.05）.Conclusions These results indicated that 1,25（OH）2D3 pretreatment enhanced the inhibitory effects of immunotherapy on allergic airway inflammation. In the treatment of allergic diseases, 1,25（OH）2D3 pretreatment may be beneficial for improving the efficacy of immunotherapy.

维生素D3对2型糖尿病小鼠轻度认知障碍的改善作用及机制研究

目的:探讨维生素D3对2型糖尿病小鼠轻度认知障碍的改善作用,并进一步探讨其可能的作用机制。方法:将雄性db/db小鼠随机分为4组:糖尿病(diabetes mellitus,DM)模型组、维生素D3低剂量组[250 IU/(kg·d)]、中剂量组[500 IU/(kg·d)]和高剂量组[1000 IU/(kg·d)],将与其匹配的db/m小鼠作为正常对照组。维生素D3各剂量组每天灌胃相应浓度的维生素D3玉米油溶液,正常对照组和DM模型组灌胃玉米油,连续饲养16周。分别于实验第0、4、8和16周检测各组小鼠空腹血糖,实验第16周进行新物体识别实验,实验结束后留取各组小鼠的海马和皮质,分别检测各组小鼠海马组织中5-羟色胺(5-hydroxytryptamine,5-HT)浓度、皮质组织中白细胞介素-18(interleukin-18,IL-18)的含量及海马组织中核苷酸结合寡聚化结构域样受体蛋白3(nucleotide binding oligomerization domain-like receptor family pyrin domain-containing protein 3,NLRP3)的蛋白表达情况。结果:与正常对照组相比,DM模型组小鼠空腹血糖值显著升高(P<0.01),新物体探索次数、探索时间和辨别指数均显著下降(P<0.05),海马组织中5-HT浓度显著下降(P<0.01),皮质组织中IL-18浓度显著升高(P<0.01),海马组织中NLRP3的阳性表达明显增加。与DM模型组相比,第8周和第16周的维生素D3中剂量和高剂量组小鼠血糖显著下降(P<0.05或P<0.01),维生素D3高剂量组小鼠新物体探索次数、探索时间和辨别指数均显著升高(P<0.05),中剂量和高剂量组小鼠海马组织中5-HT浓度显著升高(P<0.01)且皮质组织中IL-18浓度显著降低(P<0.01),各剂量组小鼠海马组织中NLRP3的阳性表达明显降低。结论:维生素D3可能通过抑制NLRP3活性降低炎症反应,进而改善2型糖尿病小鼠轻度认知障碍。

NAFLD患者血清25(OH)D3,Vaspin和AIP水平及其与疾病严重程度的相关性研究

目的 探讨非酒精性脂肪性肝病(non alcoholic fatty liver disease,NAFLD)患者血清25-羟维生素D3[25-hydroxy vitamin D3,25-(OH)D3]、内脏脂肪特异性丝氨酸蛋白酶抑制剂(Vaspin),血浆动脉粥样硬化指数(atherogenic index of plasma,AIP)水平及其与疾病严重程度之间的关系。方法 收集2021年1月~2023年1月陕西省人民医院收治的112例NAFLD患者及同期健康体检人群110例作为研究对象。根据B超检查结果进一步将NAFLD分为轻度(n=52)、中度(n=38)和重度(n=22)脂肪肝组。测量所有研究对象身高、体重、腰围(WC)和臀围,计算体质量指数(BMI)及腰臀比(WHR)。采用自动生化分析仪测定空腹血糖(FPG)、空腹胰岛素(FINS)、三酰甘油(TG)、总胆固醇(TC)、低密度脂蛋白-胆固醇(LDL-C)、高密度脂蛋白-胆固醇(HDL-C)、丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)。采用酶联免疫吸附试验(ELISA)测定25 (OH)D3和Vaspin水平,计算胰岛素抵抗指数(HOMA-IR)和AIP。分析NAFLD与健康对照组之间、不同严重程度NAFLD患者上述指标的差异。多因素Logistic回归分析影响NAFLD严重程度的因素。偏相关分析25-(OH)D3水平、Vaspin,AIP和NAFLD严重程度之间的相关性。结果 112例NAFLD患者中有78例(69.6%)患者25 (OH)D3缺乏或不足,NAFLD患者Vaspin,25 (OH)D3和AIP平均水平为7.12±2.36ng/ml,17.24±5.56ng/ml和0.68±0.13。与对照组比较,NAFLD组患者BMI,WHR,TG,TC,LDL-C,ALT,AIP和HOMA-IR升高(t=2.323,2.145,3.267,2.532,3.128,3.134,2.625,5.041),而Vaspin,25-(OH)D3水平降低(t=4.542,5.163),差异具有统计学意义(均P<0.05)。与轻、中度NAFLD患者比较,重度患者BMI,WHR,TG,ALT,AIP和HOMA-IR升高,而Vaspin,25-(OH)D3水平降低,差异具有统计学意义(F=8.645~76.742,均P<0.05)。Logistic多因素分析显示BMI(OR=1.922,95%CI:1.175~3.143),TG(OR=2.464,95%CI:1.263~4.807),AIP(OR=1.146, 95%CI:1.045~1.257)和HOMA-IR(OR=1.298,95%CI:1.046~1.612)为NAFLD危险因素,而Vaspin(OR=0.26, 95%CI:0.097~0.698)和25-(OH)D3(OR=0.271, 95%CI:0.099~0.600)为保护性因素。偏相关分析显示25-(OH)D3浓度与Vaspin水平之间呈正相关(r=0.53,P<0.01),而与AIP,ALT,AST和HOMA-IR呈负相关(r=-0.32,-0.26,-0.34,-0.27,均P <0.01)。结论 NAFLD患者血清25-(OH) D3,Vaspin低表达、AIP高表达与NAFLD相关并影响肝脏脂肪变。25-(OH)D3是NAFLD一种潜在的药物靶点,对NAFLD防治具有指导意义。

维生素D3辅助治疗小儿支气管哮喘急性发作及对外周血EOS、IRF1、ADAM8的影响

目的研究维生素D3(VD3)辅治小儿支气管哮喘(BA)急性发作及对外周血嗜酸性粒细胞(EOS)、干扰素调控因子1(IRF1)、解整合素样-金属蛋白酶8(ADAM8)的影响。方法前瞻性纳入2021年7月至2023年6月淮安市第二人民医院收治的BA急性发作患儿118例,依据随机数字表法将其分为试验组(n=59)与对照组(n=59)。对照组给予基础治疗+孟鲁司特钠+布地奈德治疗。在对照组基础上,试验组给予VD3治疗。两组均治疗7 d。观察两组临床症状消失时间,治疗前后肺功能指标[第1秒用力呼气容积(FEV1)、用力肺活量(FVC)及FEV1/FVC]、外周血炎症因子[白细胞介素(IL)-17、IL-35]水平、外周血EOS、IRF1蛋白、ADAM8蛋白水平。结果试验组胸闷、气促、喘息、肺内哮鸣音及咳嗽消失时间分别为(5.97±0.62)、(3.64±0.38)、(3.58±0.37)、(6.59±0.68)、(7.22±0.77)d,均小于对照组[(6.31±0.65)、(3.87±0.40)、(3.82±0.40)、(6.94±0.73)、(8.30±0.85)d],差异均有统计学意义(P<0.05)。治疗后,试验组FEV1、FVC及FEV1/FVC分别为(2.41±0.26)L、(3.24±0.33)L、(74.38±7.55)%,均高于对照组[(2.18±0.24)L、(3.11±0.30)L、(70.09±7.24)%],差异均有统计学意义(P<0.05)。试验组外周血IL-17水平为(44.55±4.63)pg/mL,低于对照组[(47.33±4.86)pg/mL],IL-35水平为(215.83±23.06)pg/mL,高于对照组[(202.95±21.36)pg/mL],差异均有统计学意义(P<0.05)。治疗后,试验组外周血外周血EOS水平及IRF1蛋白、ADAM8蛋白水平分别为(0.15±0.02)×10^(9)/L、(1.07±0.12)pg/mL、(1.14±0.13)pg/mL,均低于对照组[(0.16±0.02)×10^(9)/L、(1.14±0.14)pg/mL、(1.21±0.15)pg/mL],差异均有统计学意义(P<0.05)。试验组总有效率为94.92%,高于对照组(81.36%),差异有统计学意义(P<0.05)。治疗期间,两组均无显著不良反应发生。结论VD3辅助治疗小儿BA急性发作可纠正机体免疫炎症反应失衡,缓解气道炎症反应,抑制生成EOS、IRF1、ADAM8,缩短治疗时间,提高肺功能,疗效显著,安全性高。

Hepc/25(OH)D3、EPO、sTfR-F与慢性肾衰竭MHD患者肾性贫血关系及联合预测并发感染的ROC分析

目的:探讨铁调素与维生素D3比值[Hepc/25(OH)D3]、促红细胞生成素(EPO)、可溶性转铁蛋白受体/铁指数(sTfR-F)与慢性肾衰竭维持性血液透析(MHD)患者肾性贫血的关系及联合预测并发感染的价值。方法:选取2018年1月-2020年12月我院139例慢性肾衰竭MHD患者,根据透析1个月内是否并发感染分为感染组(n=32)、非感染组(n=107),比较两组Hepc/25(OH)D3、EPO、sTfR-F、血红蛋白(Hb)、转铁蛋白饱和度(TSAT)、血细胞比容(HCT),分析Hepc/25(OH)D3、EPO、sTfR-F与肾性贫血相关指标关系、感染相关影响因素及各指标预测感染的价值。结果:感染组Hepc/25(OH)D3、sTfR-F高于非感染组,EPO低于非感染组(P<0.05);感染组Hb、TSAT、HCT低于非感染组(P<0.05);Hepc/25(OH)D3、sTfR-F与Hb、TSAT、HCT呈负相关,EPO与Hb、TSAT、HCT呈正相关(P<0.05);Hepc/25(OH)D3、EPO、sTfR-F均为慢性肾衰竭MHD患者发生感染的影响因素(P<0.05);Hepc/25(OH)D3、EPO、sTfR-F预测慢性肾衰竭MHD患者并发感染的曲线下面积(AUC)分别为0.810、0.784、0.765,各指标联合预测的AUC最大,为0.864。结论:Hepc/25(OH)D3、EPO、sTfR-F与慢性肾衰竭MHD患者肾性贫血密切相关,且是感染发生的影响因素,各指标联合在预测感染发生方面具有较高应用价值。

软肝抗纤方联合恩替卡韦与维生素D3治疗乙型肝炎肝硬化代偿期患者的临床疗效及对其免疫功能的影响

目的 探讨软肝抗纤方联合恩替卡韦与维生素D3对乙型肝炎肝硬化(Hepatitis B cirrhosis, HBC)代偿期患者的临床疗效及免疫功能的影响。方法 选取2021年1月—2022年1月期间华北石油管理局总医院门诊收治的代偿期HBC患者120例,按照随机数字表法分为对照组和观察组,每组各60例。对照组予恩替卡韦加维生素D3治疗,观察组在对照组基础上联用软肝抗纤方治疗。治疗24周后,观察比较两组患者临床疗效,治疗前后肝功能指标[天门冬氨酸氨基转移酶(Aspartate aminotransferase, AST)、丙氨酸氨基转移酶(Alanine aminotransferase, ALT)、白蛋白(Albumin, ALB)和总胆红素(Total bilirubin, TBil)]、肝纤维化指标[层黏连蛋白(Laminin, LN)、透明质酸(Hyaluronic acid, HA)、Ⅲ型前胶原(Type III procollagen, PCⅢ)、Ⅳ型胶原(Type IV collagen,Ⅳ-C)]、免疫功能指标[CD4^(+)、CD8^(+),并计算CD4^(+)/CD8^(+)],评价中医证候评分与慢性肝脏疾病特异性量表(The chronic liver disease-specific questionnaire, CLDQ)评分。结果 治疗24周后两组患者中医证候评分较治疗前降低,差异有统计学意义(P<0.05);且观察组评分低于对照组,差异有统计学意义(P<0.05)。治疗24周后观察组总有效率96.67%(58/60)高于对照组83.33%(50/60),差异有统计学意义(P<0.05)。治疗24周后肝功能指标AST、ALT和TBil均较治疗前降低,ALB较治疗前升高,差异有统计学意义(P<0.05);且观察组肝功能指标ALT、AST和TBil低于对照组,ALB高于对照组,差异有统计学意义(P<0.05)。治疗24周后,两组患者肝纤维化指标LN、HA、PCⅢ和Ⅳ-C均较治疗前降低,差异有统计学意义(P<0.05);且观察组低于对照组,差异有统计学意义(P<0.05)。治疗24周后两组患者CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+)均较治疗前升高,差异有统计学意义(P<0.05);且观察组CD4^(+)、CD4^(+)/CD8^(+)高于对照组,差异有统计学意义(P<0.05);CD8^(+)与对照组比较,差异无统计学意义(P>0.05)。治疗24周后两组患者生活质量CLDQ评分均较治疗前升高,差异有统计学意义(P<0.05);且观察组高于对照组,差异有统计学意义(P<0.05)。观察组不良反应率5.00%(3/60)与对照组1.67%(1/60)比较,差异无统计学意义(P>0.05)。结论 软肝抗纤方联合恩替卡韦与维生素D3治疗HBC可提高临床疗效,调节免疫功能,改善临床症状及肝功能,改善肝纤维化。