大鼠甲状旁腺细胞经培养后再移植的实验研究

目的　研究大鼠甲状旁腺细胞经培养后再移植对其存活的影响。方法　将经胶原酶和胰蛋白酶消化的甲状旁腺细胞培养后再行移植 ,观察移植物的存活情况 ,并对移植物做透射电镜观察。结果　新鲜甲状旁腺移植组平均存活期为 (9.2 5± 3.4 5 )d ;甲状旁腺细胞培养后移植 ,移植物的存活时间为 (46 .2 5± 7.4 4 )d ,明显延长(P<0 .0 1) ,在 5 0d观察期内 ,8只鼠中有 6只的血清钙及PTH值持续在正常范围内。移植物内可见完整的甲状旁腺细胞 ,其内见丰富的粗面内质网、杆状的线粒体及分泌颗粒。结论　大鼠甲状旁腺细胞经培养后再移植可以延长移植物的存活时间 ,是治疗甲状旁腺功能低下症的一条有效途径。     

微囊化兔甲状旁腺组织异种移植治疗甲状旁腺功能低下症的实验研究

目的探讨不同部位微囊化兔甲状旁腺组织异种移植治疗大鼠甲状旁腺功能低下症的效果。方法应用微囊技术制备微囊化（海藻酸钠-钡生物微胶囊）兔甲状旁腺组织。16只去甲状旁腺的Wistar大鼠按完全随机法分为肾脂肪囊微囊组及腹腔微囊组，每组8只。分别于移植微囊化兔甲状旁腺组织后5、15、25、35、45、55及65d检测血清钙含量，并于移植后第65d取移植物行透射电镜检查。结果移植后，肾脂肪囊微囊组大鼠血清钙水平恢复到正常范围直至观察结束时，透射电镜检查显示移植物存活良好；腹腔微囊组大鼠血清钙水平恢复到正常范围，但第65d时血清钙水平下降至〈1．6mmol／L，透射电镜检查显示移植物中心坏死，仅边缘有少数存活细胞。结论微囊化兔甲状旁腺组织异种移植在不用免疫抑制剂情况下，可以在大鼠体内存活，且有功能。不同移植部位移植后随着时间的延长存在差异，肾脂肪囊在微囊化异种组织移植中是首选的移植部位。     

微囊化异种甲状旁腺组织移植的研究

目的探讨微囊化异种甲状旁腺(PTG)组织移植对Wistar大鼠甲状旁腺功能低下的治疗作用及微囊的通透性。方法 40只去甲状旁腺Wistar大鼠随机分为微囊组、非微囊组、空囊组、空白对照组。用海藻酸-钡交联微囊包裹兔甲状旁腺组织,移植至Wistar大鼠肾包囊。移植后每隔2周取血测血钙,移植后第16周取移植物进行透射电镜检查及T淋巴细胞、大鼠IgG抗体的免疫组织化学染色。结果微囊组移植后第4周血清钙由(1．62±0．04)mmol／L恢复至正常水平 (2．2～2．6)mmol／L,9例维持至观察期结束(P<0．01),非微囊组、空囊组及空白对照组血清钙差异无统计学意义(P>0．05)。第16周取出移植物检测显示移植物活性良好,微囊周围可见较多T 淋巴细胞浸润,囊壁及囊内IgG抗体染色阳性。结论海藻酸-钡交联微囊可对甲状旁腺组织起到有效的保护作用,使甲状旁腺组织较长时间存活并发挥正常功能。但海藻酸-钡交联微囊并未减轻受体免疫排斥反应的激活且未有效的免疫隔离IgG抗体。     

去甲状旁腺模型鼠的制备

目的探讨去甲状旁腺(PTG)模型鼠的制备方法,为PTG移植奠定实验基础。方法手术切取PTG,并行病理学检查。对术前和术后2、5、10、15和30d的大鼠血清钙和甲状旁腺激素(PTH)进行测定。结果8只鼠的PTG被成功切除,切除率为80%(8/10)。假手术组手术前、后血清钙和PTH水平无明显变化(P>0.05)。PTG切除组术后2、5、10、15和30d血清钙和PTH水平均明显低于术前,并明显低于同时相假手术组水平(P<0.01)。结论准确切取大鼠PTG,可成功制备去PTG模型鼠。术后10d可作为异体移植的适宜时间。     

甲状旁腺细胞与睾丸Sertoli细胞共同移植产生免疫赦免的研究

目的　研究甲状旁腺细胞与睾丸细胞共同移植是否产生免疫赦免。方法　随机将SD大鼠分为以下四组 :1组为单纯甲状旁腺细胞移植组 ;2组为甲状旁腺细胞 1× 10 6个睾丸Sertoli细胞移植组 ;3组为甲状旁腺细胞 2× 10 6个睾丸Sertoli细胞移植组 ;4组为甲状旁腺细胞 4× 10 6个睾丸Sertoli细胞移植组。观察移植物的存活情况 ;体外检测移植物内细胞成分及淋巴细胞凋亡情况。结果　 1组平均存活期为 (17.2 2± 3.6 3)d ;2、3、4组存活时间延长 ,分别为 (19.4 4± 4 .6 4 )d、(32 .2 2± 6 .6 7)d和 (48.80± 3.33)d。 4组在 5 0d观察期内 ,多数鼠的血清钙及甲状旁腺激素 (PTH)值维持在正常范围内 (P <0 .0 1)。移植物内可见表达FasL的睾丸细胞、甲状旁腺细胞及凋亡的淋巴细胞。FasL的表达及凋亡指数随着移植的Sertoli细胞数增加而增加。结论　睾丸Sertoli细胞FasL的表达诱导了浸润的活化淋巴细胞凋亡 ,使共同移植的甲状旁腺细胞存活期延长。     

微囊化新生猪甲状旁腺细胞异种移植的实验研究

目的 探讨微囊化新生猪甲状旁腺细胞异种移植治疗大鼠甲状旁腺功能低下症的效果。方法 应用微囊化技术，制备微囊化（海藻酸钠－聚赖氨酸－海藻酸钠生物微胶囊）新生猪甲状旁腺细胞，32只去甲状旁腺的Wistar大鼠随机分成微囊组、非微囊组、空囊组和对照组，分别移植微囊化新生猪甲状旁腺细胞、甲状旁腺细胞、空微囊及生理盐水。移植后监测血钙及甲状旁腺素水平40周，40周后回收移植物，透射电镜检查。结果 移植后，微囊组大鼠血钙及甲状旁腺素水平恢复到正常范围内，直至观察结束时（40周），透射电镜检查显示移植物存活良好；非微囊组、空囊组和对照组大鼠的血钙及甲状旁腺素水平无改善。结论 微囊化新生猪甲状旁腺细胞异种移植在不用免疫抑制剂情况下，可以在大鼠体内存活，且有功能；海藻酸钠－聚赖氨酸－海藻酸钠生物微胶囊对免疫活性细胞及抗体具有屏蔽作用。     

异体甲状旁腺脑室内移植治疗甲状旁腺机能减退症

应用异体甲状旁腺进行脑室内移植治疗甲状旁腺机能减退症６例，疗效满意，术后临床症状消失或明显减轻，连续监测血清钙、血清磷和尿钙、尿磷均证实移植物在脑室内存活并有正常的生理功能。随访７－２２个月病情均无复发。本文对脑室内组织移植的手术方法，胎儿甲状旁腺的取材及术后可能发生的并发症进行了讨论。     

环胞霉素在大鼠甲状旁腺同种异体移植中的应用

为了研究环胞霉素不同剂量对甲状旁腺移植后存活时间的影响，选用纯系Ｆｉｓｈｅｒ３４４大白鼠４２只，分为三组，分别注射剂量的环胞霉素，将移植模型的大鼠，从接受供体大鼠的甲状旁腺开始，移植后第５，１０，１５，２０，２５天分别取尾部血，测血清钙值，结果发现：不同的给药方式能影响移植物的发生排斥反应时间，定期短疗程多次强化注射比只注射一个疗程的ｃｙｃｌｏｓｐｏｒｉｎｅ效果显著。     

人胚甲状旁腺细胞移植治疗甲状旁腺功能低下症

目的　探讨人胚甲状旁腺细胞移植治疗甲状旁腺功能减退症的临床意义。　方法　将培养的人胚甲状旁腺细胞在B超引导下移植到 6例甲状旁腺功能减退症患者的肾周围脂脂囊中。应用放射免疫方法测定患者术前、术后血中甲状旁腺激素 (PTH)及钙水平 ,并进行自身对照 ,对疗效进行评价。　结果　 6例患者在接受人胚甲状旁腺细胞移植前 ,血钙及PTH平均水平分别为 (1 6 3±0 12 )mmol/L及 (1 36± 0 2 1)ng/L ;接受移植 3d后分别为 (1 77± 0 2 2 )mmol/L及 (9 5 3± 2 2 1)ng/L ,两者差异有显著性意义 (P <0 0 1) ;6d后达正常水平 ,临床症状逐渐减轻至消失 ;术后观察 9～ 12个月病情保持稳定。　结论　培养的人胚甲状旁腺细胞肾周围脂肪囊内移植是治疗甲状旁腺功能减退症的一种较理想的新方法     

纳米碳对甲状旁腺切除加自体移植术中甲状旁腺识别作用探讨

目的探讨纳米碳对甲状旁腺切除加自体移植术中甲状旁腺识别作用。方法尿毒症继发性甲状旁腺功能亢进病人74例,分为观察组(39例)和对照组(35例)。观察组采取纳米碳术中示踪的甲状旁腺切除加自体移植术;对照组采取传统的甲状旁腺切除加自体移植术;随访6个月,观察手术结果、病理诊断、血清甲状旁腺激素(PTH)、血钙水平、并发症及复发情况。结果两组术后症状均较术前明显改善;观察组甲状旁腺组织切除准确率为100%,对照组为86.18%,差异有统计学意义(P0.05);两组手术切口长度、术中出血量差异均无统计学意义(P0.05);观察组手术时间、术后住院时间短于对照组,差异有统计学意义(P0.05);观察组术后1天及6个月时血清PTH、血钙水平低于对照组,差异有统计学意义(P0.05);两组均未见喉返神经损伤。观察组术后出现低血钙20例,对照组术后均出现不同程度的低血钙,经对症处理后缓解;随访期间,观察组复发率为0,对照组为11.43%,差异有统计学意义(P0.05)。结论纳米碳对甲状旁腺切除加自体移植术中异常甲状旁腺有识别作用,有助于保护甲状腺功能和喉返神经。     

Sevelamer hydrochloride reverses parathyroid gland enlargement via regression of cell hypertrophy but not apoptosis in rats with chronic renal insufficiency.

BACKGROUND: Dietary phosphate restriction suppresses parathyroid hormone (PTH) secretion, synthesis, and parathyroid cell proliferation in experimental animals with chronic renal insufficiency (CRI), independently of serum calcium and 1,25(OH)2D3 levels. This study was conducted to examine whether sevelamer hydrochloride (sevelamer), a metal-free phosphate binder, could regress an advanced parathyroid gland (PTG) hyperplasia and enlargement in rats with CRI. METHODS: Male Sprague-Dawley rats were fed a diet containing adenine for 6 weeks to establish CRI. Normal rats and adenine-treated rats were sacrificed to obtain the PTG (baseline group). The adenine diet was changed to a normal diet or diet containing 1 or 3% sevelamer for another 4 weeks. Time course changes of serum levels of calcium, phosphorus, and PTH were measured. At the end of the study, the PTG was weighed and examined histologically. RESULTS: Adenine-treated rats developed severe CRI with marked elevation of serum phosphorus and PTH. The PTG weight markedly increased with enlarged cell volume (i.e. cell hypertrophy) at baseline. Sevelamer treatment rapidly lowered serum phosphorus and PTH levels within 6 days, and after 4 weeks, reduced the PTG weight by 38% compared to adenine-treated rats at baseline. The reduction in PTG weight was due to regression of cell hypertrophy, but not to decreased cell number by apoptosis. Decreased expression of calcium receptor in the PTG at baseline was partially recovered by the sevelamer treatment. CONCLUSIONS: The sevelamer treatment can reduce the PTG weight with a reduction in serum PTH levels via regression of cell hypertrophy but not apoptosis in rats with CRI. Reduced PTG function might contribute to the regression of cell hypertrophy.     

Hyperplasia of the parathyroid gland without secondary hyperparathyroidism

BACKGROUND: Low dietary phosphorus (P) prevents parathyroid gland (PTG) hyperplasia and the development of secondary hyperparathyroidism (SH) in uremic rats. The present study explores the effects of P restriction on parathyroid hormone (PTH) synthesis and secretion and PT cell growth in rats with established SH and PTG hyperplasia. METHODS: Normal and 5/6 nephrectomized rats were fed a high P (0.8%) diet. After two weeks, the normal rats and half of the uremic rats were sacrificed (U-HP) while the remaining uremic rats were switched to a low P (0.2%) diet (U-HP-LP). RESULTS: High dietary P induced a significant increase in serum P, PTH, and PTG weight, but not ionized calcium compared to normal animals fed the same diet (N-HP). P restriction returned serum P and PTH to normal levels by one week. In contrast, PTG size did not regress and glands remained enlarged for up to eight weeks with no evidence of apoptosis. Ribonuclease protection assay and metabolic labeling studies demonstrated similar PTH/actin mRNA ratios and 35S-labeled PTH among the three groups. Intracellular intact PTH was higher in U-HP and U-HP-LP rats compared to N-HP animals with no differences between the two uremic groups. PTG-PTH content correlated only with PTG weight, and serum PTH only with serum P. The PTG secretory response to calcium remained intact. CONCLUSIONS: In established chief-cell hyperplasia, P restriction restores normal serum PTH levels without affecting PTG hyperplasia, PTH synthesis, PTG cytosolic PTH or the PTH secretory response to calcium, suggesting an impaired exocytosis of PTH.     

Sevelamer hydrochloride (Renagel), a non-calcaemic phosphate binder, arrests parathyroid gland hyperplasia in rats with progressive chronic renal insufficiency.

BACKGROUND: It has been demonstrated that dietary phosphate restriction suppresses parathyroid hormone (PTH) secretion and parathyroid cell proliferation in experimental animals with chronic renal insufficiency (CRI) independently of serum calcium and 1,25(OH)(2)D3 levels. This study was conducted to examine whether sevelamer hydrochloride (Renagel); hereafter referred to as sevelamer), a non-calcaemic phosphate binder could inhibit the parathyroid gland (PTG) hyperplasia in rats with progressive CRI. METHODS: Male Sprague-Dawley rats were injected twice with low doses of adriamycin (ADR). Two weeks after the last injection of ADR, rats were fed a diet containing 1 or 3% sevelamer for 84 days. Time course changes of serum levels of calcium, phosphorus, and PTH were measured. At the end of study, serum 1,25(OH)(2)D3 levels were measured and the maximal two-dimension area of the PTG in paraffin section was calculated using an imaging analyser. RESULTS: Dietary sevelamer treatment inhibited the elevations of serum phosphorus, calciumxphosphorus product, and PTH levels that occurred as the study progressed. Sevelamer also suppressed maximal PTG area and there existed positive strong correlation between maximal PTG area and serum PTH levels at the end of the study. Serum phosphorus levels positively correlated well with serum PTH levels and maximal PTG area. In contrast, serum calcium or 1,25(OH)(2)D3 levels did not show any correlation with serum PTH levels and maximal PTG area. CONCLUSIONS: Sevelamer treatment arrested hyperphosphataemia and PTG hyperplasia accompanied by the elevation of serum PTH levels. The correlation analysis suggests that reduced serum phosphorus levels contributed to the suppression of PTG hyperplasia and resulted in the reduction of PTH levels in this animal model after the sevelamer treatment. The management of phosphorus control started from early stage of CRI could prevent PTG hyperplasia and facilitate later management of secondary hyperparathyroidism.     

Effects of Cerivastatin on Vascularized Allogenic Bone Transplantation in Rats Treated with Cyclosporine A

The aim of the present study was to investigate the effects of oral cerivastatin (0.1 mg/kg/day) on vascularized allogenic transplanted bone that is treated with cyclosporine A (CsA) (10 mg/kg/day) and on vascularized isogenic transplanted bone that is not treated with CsA. Allogenic transplantation was performed on 12-week-old male DA rats with the major histocompatibility antigen (MHC) RT1a (as the donor) and age-matched male Lewis rats with MHC RT1l (as the recipient), and isogenic transplantation was performed on 12-week-old male Lewis rats. After transplantations, 20 rats (10 rats in each transplantation) were randomized into four groups to receive the following treatment for 16 weeks: (1) CsA plus cerivastatin vehicle or (2) CsA plus cerivastatin in the allogenic transplanted rats, and (3) CsA vehicle plus cerivastatin vehicle or (4) CsA vehicle plus cerivastatin in the isogenic transplanted rats. Bone biochemical markers, mineral density, and strength were measured at the end of the study period. Serum levels of osteocalcin (OC) and parathyroid hormone (PTH) and urinary deoxypyridinoline (DPD) level were higher in the allogenic transplanted rats than in the isogenic transplanted rats. In the allogenic transplanted rats, the cerivastatin treatment decreased urinary DPD levels, but not serum OC nor PTH levels. Furthermore, the cerivastatin treatment improved bone mineral density of the allogenic transplanted bones and bone strength of the allogenic reconstructed bones. In contrast, no effect of the cerivastatin treatment was observed in the isogenic transplanted rats. These results suggest that the cerivastatin treatment improves CsA-induced high-turnover osteopenia mainly through the inhibition of bone resorption.     

Successful kidney transplantation reduces hyperplastic parathyroid gland

INTRODUCTION: In dialysis patients, the parathyroid glands (PTGs) may increase progressively, producing abnormal bone metabolism. Changes in PTG volume among patients with hyperplastic PTGs are not well known after kidney transplantation. This study investigated PTG volume by ultrasound (US). METHODS: US of PTG was performed immediately (US-0) and 12 months after (US-12) transplantation to identify glands in all recipients. We calculated the percentage reduction in PTG volume (R%PTG). We declared it significant when it was > or =35%. Bone biochemical markers and renal function were recorded sequentially. RESULTS: Among engrafted patients, parathyroid US-0 was performed in 47 and US-0 and US-12 in 36. Some visible gland was observed upon US-0 in 13 recipients, a group that showed higher pretransplantation parathyroid hormone (PTH) levels than the remaining 34 patients with no visible glands (627 +/- 360.0 vs 280 +/- 240.9 pg/mL; P < .05). Of 36 recipients with US-0 and US-12, the baseline study identified PTGs in 12 patients (p+ group), while the remaining 24 had no identified glands (p- group). In the p+ group, no PTG, at US-12 were visible in four patients, and a significant R%PTG was observed in three at this time, representing a reduction in gland volume after transplantation among 58.3% of p+ patients. There was a progressive reduction in PTH among both groups. Patients with glandular volume reduction displayed better renal function: serum creatinine 1.7 +/- .79 versus 2.9 +/- .74 mg/dL (P < .05). CONCLUSIONS: Transplantation reversed hyperparathyroidism and PTG volume among recipients who achieved nearly normal renal function.     

Early stage of urolithiasis formation in experimental hyperparathyroidism

PURPOSE: We have previously noted marked acceleration in the proliferative activity of parathyroid cells in rats with spontaneous hypercholesterolemia and secondary hyperparathyroidism. Using this proliferative potential we investigated whether transplantation of these enlarged parathyroids into normal rats would induce hyperparathyroidism and renal stones. MATERIALS AND METHODS: We used 26-week-old male rats with spontaneous hypercholesterolemia as donors, and 5-week-old normal male Sprague-Dawley rats and rats with spontaneous hypercholesterolemia as recipients. Enlarged parathyroid glands were transplanted into group 1--Sprague-Dawley rats with no treatment, group 2--Sprague-Dawley rats that received FK-506 as an immuno-suppressor, group 3--rats with spontaneous hypercholesterolemia rats that underwent parathyroidectomy plus FK-506 administration and group 4--Sprague-Dawley rats that underwent parathyroidectomy plus FK-506 administration. Parathyroidectomy was performed in recipients before transplantation to ensure a low calcium condition. RESULTS: Grafts were rejected within 11 and 15 weeks in groups 1 and 2, respectively. In group 3, 78% of the grafts were successful even after 19 weeks. In group 4 graft survival was 30% at 15 weeks with complete rejection at 19 weeks. In group 3 gradually elevated serum parathyroid hormone was observed as well as stone plaques containing calcium oxalate and calcium phosphate in renal tubules located mainly in the corticomedullary junction. An increased number of plaques was associated with higher parathyroid hormone. CONCLUSIONS: Our study shows that transplanted parathyroid glands function with an immunosuppressive agent and the maintenance of hypocalcemic conditions, and they secrete sufficient parathyroid hormone to demonstrate hyperparathyroidism. Plaque in these kidneys indicates an early stage of urolithiasis caused by hyperparathyroidism.     

Biochemical and cellular effects of direct maxacalcitol injection into parathyroid gland in uremic rats

The most important etiological factors of resistance to medical treatments for secondary hyperparathyroidism are the decreased contents of the vitamin D receptor (VDR) and Ca-sensing receptor (CaSR) in parathyroid cells and a severely swollen parathyroid gland (PTG) as a result of hyperplasia. The effects of direct maxacalcitol (OCT) injection into PTG in terms of these factors were investigated in this study. The PTG of Sprague-Dawley rats that were 5/6 nephrectomized and fed a high-phosphate diet were treated by a direct injection of OCT (DI-OCT) or vehicle (DI-vehicle). The changes in serum intact parathyroid hormone (PTH), Ca(2+), and phosphorus levels, in VDR and CaSR expression levels in parathyroid cells, and in Ca(2+)-PTH curves were examined. Apoptosis was analyzed by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling method and DNA electrophoresis for PTG. DI-OCT markedly decreased serum intact PTH level, and a significant difference in this level between DI-OCT and DI-vehicle was observed. However, serum Ca(2+) and phosphorus levels did not changed markedly in both groups. The upregulations of both VDR and CaSR, the clear shift to the left downward in the Ca(2+)-PTH curve, and the induction of apoptosis after DI-OCT were observed. These findings were not observed in the DI-vehicle-treated rats. Moreover, these effects of DI-OCT were confirmed by the DI-OCT into one PTG and DI-vehicle alone into another PTG in the same rat. DI-OCT may introduce simultaneous VDR and CaSR upregulations and the regression of hyperplastic PTG, and these effects may provide a strategy for strongly suppressing PTH levels in very severe secondary hyperparathyroidism.     

Assessment of parathyroid autotransplantation for preservation of parathyroid function after total thyroidectomy

BACKGROUND: Hypoparathyroidism with permanent hypocalcemia is a well-recognized complication after thyroid surgery. AIM: This study was conducted to assess the role of immediate parathyroid autotransplantation in the preservation of parathyroid function after total thyroidectomy. PATIENTS AND METHODS: Twenty-eight patients had autotransplantation of parathyroid glands resected or devascularized during total thyroidectomy. Data were collected prospectively regarding demographics, indication for surgery, operative procedure, pathologic diagnosis, number of glands transplanted, and subsequent course. Thyroid nodules were evaluated by ultrasonography, radionuclide scanning, and/or fine-needle aspiration cytology. All patients had serum ionized calcium, phosphorus, and intact parathyroid hormone (PTH) levels measured preoperatively and monitored regularly postoperatively for a period of 14 weeks and again at 6 months after operation. Patients were categorized into three groups according to the number of glands transplanted: one (group 1, n = 6), two (group 2, n = 14), or three glands (group 3, n = 8). In three other volunteers, one parathyroid gland was transplanted in the brachioradialis and subjected to electron microscopy 1, 2, and 4 weeks after transplantation. RESULTS: Total thyroidectomy was performed for malignant disease in 16 patients (57.1%) and for benign disease in 12 (42.9%) patients. All patients reverted to asymptomatic normocalcemia without the need for any medications within 4 to 14 weeks. Normal levels of serum markers were regained slower when one gland was transplanted compared with two or three glands (P <.01). Electron microscopic examination showed evidence of ischemic degeneration in the transplanted tissues 1 week postoperatively. Regeneration started by the second week and coincided with normalization of PTH levels. Optimum resting and nearly normal status of parathyroid tissue was achieved by the fourth week. CONCLUSIONS: This study showed that active PTH production coincides with regeneration of parathyroid cells and that autotransplantation of at least two resected or devascularized glands during total thyroidectomy nearly eliminates permanent postoperative hypoparathyroidism, thus improving the safety of total thyroidectomy performed for malignant or benign disease.