聚乙二醇化重组人生长激素对去垂体幼鼠胰岛分泌的影响及其机制

目的观测长效生长激素聚乙二醇化重组人生长激素(PEG-rhGH)是否引起胰岛素抵抗及其与短效生长激素注射用重组人生长激素(rhGH)的差异。方法 3周龄SD雄性幼鼠106只,随机取88只进行去垂体手术造模,取成模的幼鼠54只分为模型组、rhGH组、PEG-rhGH组,另18只假手术组为对照组。分别给予生理盐水(0.25 mg·kg-1·d-1)、rhGH(0.25mg·kg-1·d-1)、PEG-rhGH(1.4 mg·kg-1·周-1)处理。4周后进行糖耐量实验和胰岛素释放试验,计算胰岛素曲线面积与葡萄糖曲线面积比值,胰岛素稳态模型(HOMA-IR);检测血清生长抑素水平;免疫组织化学法检测胰岛胰岛素(INS)、胰高血糖素(GLU)、生长抑素(SS)、胰多肽(PP)。结果 PEG-rhGH组HOMA-IR较对照组、模型组低(P<0.05),与rhGH组比较无差异。AUCI/AUCG值组间比较显示PEG-rhGH组高于rhGH组,但无统计学差异。PEG-rhGH组GLU蛋白表达与模型组表达无差异。PEG-rhGH组INS表达较模型组表达上升(P<0.05)。PEG-rhGH组SS、PP表达与rhGH无差异。血清SS各组间均无明显差异。结论未观察到PEG-rhGH引起去垂体大鼠胰岛素抵抗和胰岛β细胞分泌能力下降,对胰岛分泌蛋白的表达无明显影响。     

重组人生长激素对大鼠急性胰腺炎的治疗作用

目的观察重组人生长激素(rhGH)对大鼠急性胰腺炎(AP)的治疗并探讨其机制。方法ipE.coli复制大鼠AP模型。42只健康♀SD大鼠随机分为对照组(C组)、急性胰腺炎组(AP组)及rhGH治疗组(T组)。AP、T组再按观察时点分为1、3 d两个亚组。C组ip 15 m.lkg-1生理盐水;AP组ip 15 m.lkg-1E.coli(1×1010CFU.L-1)后立即im生理盐水;T组于ipE.coli后立即im 2.25 U.kg-.1d-1rhGH。观察胰腺的病变程度并进行评分,测定血清淀粉酶的活性,免疫组化检测胰腺组织中ICAM-1蛋白的表达及NF-κB核的阳性率。结果AP两亚组胰腺组织病理评分、血清淀粉酶活性、ICAM-1蛋白表达及NF-κB核阳性率均明显高于C组水平(P<0.05);rhGH治疗能明显降低上述指标水平,尤以T3d组降低最为显著。结论rh-GH治疗能明显减轻大鼠急性胰腺炎的损伤程度,其机制可能与降低NF-κB活化程度及ICAM-1蛋白的表达水平等有关。     

重组人生长激素对败血症大鼠炎症调控网络的影响

目的 观察重组人生长激素(rhGH)对败血症大鼠炎症调控网络的影响.方法 采用ip E.coli的方法复制大鼠败血症模型.42只♀ SD大鼠随机分为正常对照组(C组)、败血症组(S组)及rhGH治疗组(T组).S、T组又依观察时点再分为1d、3 d两个亚组.观测血浆肿瘤坏死因子α(TNFα)、白细胞介素-10(IL-10)、一氧化氮(NO)水平及诱生型一氧化氮合酶(iNOS)的表达情况.结果 S1d组及T1d组血浆TNFα浓度均明显高于C组水平;S3d组血浆TNFα浓度接近C组水平,T3d组血浆TNFα浓度明显低于C组水平.S组血浆IL-10水平在各时点均显著低于C、T组水平,T组血浆IL-10水平在各时点与C组相比均无明显差异.S组血浆NO浓度及iNOS活力明显高于C组水平及T组水平,T组血浆NO浓度及iNOS活力仅于1 d时,明显高于C组水平.结论 rhGH在减少败血症大鼠血浆TNFα、NO等促炎介质生成与释放的同时,也能有效地维持IL-10的抗炎介质水平,提示rhGH治疗可能有助于维持败血症时炎症调控网络的平衡.     

重组人生长激素治疗新生儿败血症的临床研究

目的观察重组人生长激素治疗败血症患儿的临床疗效及对肌酸激酶同工酶(CK-MB)、肌酸磷酸激酶(CK)和血清炎症因子的影响。方法 2011年2月—2014年5月武警河北总队医院收治的败血症患儿80例,随机分为对照组和治疗组,每组40例。对照组给予综合治疗,同时给予抗生素治疗。治疗组患者入院第2天腹部sc注射用重组人生长激素,0.1 U/kg,隔日1次,其他治疗同对照组。两组均连续治疗10 d。比较两组的临床疗效,同时比较两组治疗前后CK-MB、CPK、中性粒细胞凝胶酶载脂蛋白(NGAL)、肿瘤坏死因子-α(TNF-α)以及胰岛素样生长因子-1(IGF-1)的变化。结果对照组和治疗组总有效率分别为77.5%、90.0%,两组比较差异有统计学意义(P0.05)。治疗后,两组CK、CK-MB、NGAL、TNF-α、IGF-1水平均较治疗前明显下降,同组比较差异有统计学意义(P0.05);且治疗组的改善程度优于对照组,两组比较差异有统计学意义(P0.05)。结论重组人生长激素治疗新生儿败血症有较好的临床疗效,可降低患儿心肌损伤,减轻全身炎症反应。     

异丙酚对大鼠脑内nNOS表达及NO含量的影响

目的通过观察异丙酚对大鼠海马以及基底前脑内神经型一氧化氮合酶(nNOS)的表达和一氧化氮(NO)释放量的影响,探讨异丙酚的中枢麻醉机制。方法选择正常雄性Wistar大鼠30只随机分为三组,分别腹腔注射异丙酚50mg/kg(P50组)、100mg/kg(P100组)及生理盐水10mL/kg(NS组)。10min后,各组大鼠经主动脉灌注生理盐水,随机选取5只取新鲜鼠脑,分离海马及基底前脑,分光光度法测其NO释放量;每组另5只大鼠继续灌注固定液后取脑,免疫组化检测海马及基底前脑内nNOS的表达。结果与对照组NS组相比,海马及基底前脑内nNOS表达量、NO释放量都显著减少(P<0.05)。结论异丙酚可能通过抑制海马及基底前脑内nNOS表达,使NO的释放减少而发挥麻醉作用。     

生长激素释放抑制激素联合重组人生长激素治疗重症急性胰腺炎临床研究

目的：探讨生长激素释放抑制激素( SS)联合重组人生长激素( rhGH)治疗重症急性胰腺炎( SAP)的效果。方法对2013年7月—2015年7月收治的SAP 51例的临床资料进行回顾性分析,根据治疗方案分为观察组30例和对照组21例。对照组给予SS微量泵静脉注射,24 h维持；观察组于应用SS第4天注射rhGH。观察两组恢复情况、中转手术率、严重并发症发生率及病死率,并记录不良反应发生情况,检测两组炎性细胞水平、T淋巴细胞水平及肠黏膜屏障功能指标。结果观察组腹痛消失时间、住ICU时间、住院时间均短于对照组( P<0．05)。两组治疗后肿瘤坏死因子-α(TNF-α)、白介素-1(IL-1)、白介素-6(IL-6)、高迁移率蛋白-1(HMGB-1)、二胺氧化酶、D-乳酸水平均低于治疗前,且观察组低于对照组( P<0．05)。两组治疗后CD4+、CD4+/CD8+高于治疗前,且观察组高于对照组(P<0．05)。观察组中转手术率、严重并发症发生率和病死率低于对照组(P<0．05)。观察组1例死于多器官功能衰竭,对照组死于多器官功能衰竭和呼吸衰竭各1例。两组均未发生严重不良反应。结论 SS联合rhGH治疗SAP能够减轻炎症水平,提高机体免疫力,降低中转手术率和严重并发症发生率。     

重组人生长激素对大鼠肠道屏障功能的影响

目的 探讨重组人生长激素(rhGH)对大鼠肠道屏障功能的影响.方法 45只SD大鼠随机均分为3组:A组不予处理;B组肌注生理盐水;C组用肌注rhGH 1.25 U· kg-1·d-1治疗.1周后测量各组大鼠小肠绒毛高度、肠腺隐窝深度、黏膜厚度以及绒毛表面积和小肠黏膜CD3+、CD4+、CD8+和IgA+细胞数.结果 C组的小肠绒毛高度、肠腺隐窝深度、黏膜厚度以及绒毛表面积等形态学相关参数明显好于A组和B组(P＜0.05);C组小肠黏膜CD3+、CD4+、CD8+和IgA+细胞数高于A组和B组(P＜0.05);A组与B组上述指标的差异无统计学意义.结论 rhGH可以增强小肠黏膜屏障功能及免疫屏障功能.     

重组人生长激素治疗肝硬化低清蛋白血症

目的:观察重组人生长激素(rhGH)治疗肝硬化引起的低清(白)蛋白血症的临床疗效。方法:选择血清清蛋白低于30g·L-1的肝硬化病人46例,男性39例,女性7例,分为治疗组、对照组各23例。治疗组在常规保肝及对症治疗基础上,给予rhGH,4IU,皮下注射,每日1次,连续14d;对照组仅给予常规保肝及对症治疗,14d为一个疗程。检测2组病人治疗前及治疗结束d1和4wk后血清清蛋白及其他肝功能指标变化,并进行临床观察。结果:治疗组治疗前血清清蛋白(25±s3)g·L-1,治疗结束d1血清清蛋白(31±3)g·L-1(P<0.01),4wk后血清清蛋白(33±3)g·L-1(P<0.01);对照组治疗前血清清蛋白(25±3)g·L-1,治疗结束d1血清清蛋白(25±3)g·L-1,4wk后血清清蛋白(24.4±2.6)g·L-1(P>0.05),2组治疗结束d1及4wk后血清清蛋白相比,差异有非常显著意义(P<0.01)。治疗结束d1及4wk内治疗组比对照组病人精神状态、食欲、尿量、腹腔积液、肝功能等均明显改善且持续稳定,有显著差异(P<0.05)。无明显不良反应。结论:rhGH可以增加肝硬化病人的血清清蛋白的水平,改善营养状况,无明显不良反应。     

重组人生长激素治疗青春期前特发性矮小儿童

目的:观察重组人生长激素(rhGH)治疗青春期前特发性矮小的疗效。方法:24例特发性矮小病儿分为2组,治疗组女孩10例,男孩2例,起始治疗年龄为(10.1±s1.9)a,予rhGH0.10～0.13U·kg-1·d-1,治疗时间(0.8±0.3)a。对照组女孩10例,男孩2例,起始观察年龄(9.9±1.7)a,未予rhGH治疗,观察时间(1.1±0.5)a。结果:治疗组治疗后生长的速率为(8.7±1.9)cm·a-1,对照组为(5.0±0.7)cm·a-1,2组差异有非常显著意义(P<0.01)。治疗组对骨龄的身高标准差值(HtSDSBA)增加0.4±0.4,预测成人身高(PHt)增加了(3.1±0.9)cm;对照组HtSDSBA减少0.3±0.5,PHt降低(0.9±1.7)cm,2组比较差异均有非常显著意义(P<0.01)。结论:rhGH对青春期前特发性矮小儿童的生长有促进作用,能改善其PHt。     

基因重组人生长激素治疗青春期前特发性生长激素缺乏症临床试验

目的:评价基因重组人生长激素(rhGH)治疗青春期前特发性生长激素缺乏症(IGHD)患儿6个月的疗效和安全性。方法:对49例IGHD患儿进行rhGH治疗,每晚睡前皮下注射0．5IU·kg~(-1),每周分5次注射,共26周。评价治疗前后身高、生长速度、身高标准差计数(SDS)等指标。结果:在rhGH治疗期间,46例患儿生长速度由每年(2．5±1．0)cm提高到(11．5±2．5)cm(P＜0．005)。身高SDS由治疗前(-4．2±1．8)增为(-3．7±1．8)。同时患儿13和26周血清胰岛素样生长因子(IGF-1)和IGF结合蛋白(IGFBP,)水平均较治疗前明显升高(P＜0．05)。其体重与骨龄无明显变化。治疗后共有15．7%患儿出现甲状腺功能降低,抗hGH抗体阳性率为21．7%,但所有这些现象均未影响患者体格的线性增长。在治疗期间所有患者肝肾功能、血尿常规和代谢性指标(如血糖和血脂水平)等均保持在正常范围。结论:rhGH是治疗IGHD安全有效的药物。     

重组人促红细胞生成素对大鼠肝脏缺血-再灌注损伤的保护作用

目的探讨重组人促红细胞生成素(rHuEPO)对大鼠肝脏缺血-再灌注(I-R)损伤的保护作用。方法将30只大鼠随机均分为假手术组(A组)、I-R模型组(B组)和rHuEPO 5000IU/kg处理组(C组)。建立70%大鼠肝脏I-R损伤模型,检测各组血清ALT、AST水平,HE染色观察组织学变化,Western blot及RT-PCR检测肝脏诱导型一氧化氮合酶(iNOS)和内皮型一氧化氮合酶(eNOS)的表达。结果与B组相比,C组iNOS mRNA和蛋白表达、ALT和AST水平均明显减少(P<0.05或P<0.01),肝细胞水肿、坏死减轻;而B组和C组间eNOS mRNA和蛋白表达差异无统计学意义(P>0.05)。结论 rHuEPO可能通过抑制iNOS表达,减轻大鼠肝脏I-R损伤。     

重组人生长素治疗大面积烧伤伴重度吸入性损伤的疗效观察

目的 探讨人重组生长素（rhGH）治疗大面积烧伤伴重度吸人性损伤的疗效。方法40例大面积烧伤伴重度吸人性损伤患者被随机分为治疗组及对照组，治疗组行烧伤常规治疗＋rhGH治疗，对照组仅行烧伤常规治疗．观察两组治疗后营养状况（包括左上臂肌围、血红蛋白、总蛋白、白蛋白等）、创面愈合时间、机械通气时间及住院日。结果 治疗后两组营养状况差异有统计学意义（P〈0．05），治疗组植皮区、供皮区愈合时间．机械通气时间及住院日均优于对照组（P〈0．05）。结论rhGH能促进创面愈合，改善营养状况，缩短机械通气时间。     

重组人脑利钠肽治疗急性肺损伤的临床效果及对 NLRP3/6、白细胞介素水平的影响

目的 探讨重组人脑利钠肽治疗急性肺损伤的临床效果及对NLRP3/6及白细胞介素水平的影响.方法 选取2016年9月至2018年9月江南大学附属医院诊治的急性肺损伤病人80例.按照治疗方法采用随机数字表法分为对照组和观察组,每组40例.对照组给予常规治疗,观察组在对照组基础上给予重组人脑利钠肽治疗.分析两组治疗前后Murray评分及氧合指数的变化,并检测治疗前后血清中NLRP3/6及白细胞介素(IL)水平的变化.结果 对照组和观察组治疗后Murray评分[(2.36±0.43)分、(1.19±0.33)分]、NLRP3[(94.6±10.3)pg/mL、(77.7±11.3)pg/mL]、IL-1β[(38.6±12.3)pg/mL、(30.7±12.5)pg/mL]、IL-2[(42.4±12.5)pg/mL、(36.5±10.4)pg/mL]及IL-6[(39.5±13.7)pg/mL、(30.8±11.6)pg/mL]水平明显降低(P<0.05),而氧合指数[(283.4±18.2)、(309.4±24.3)]和NLRP6[(88.9±15.1)pg/mL、(97.8±13.3)pg/mL]明显升高(P<0.05),且观察组治疗后上述项目变化更显著(P<0.05);NLRP6与急性肺损伤呈负相关性而NLRP3、IL-1β、IL-2及IL-6水平与急性肺损伤呈正相关性.结论 重组人脑利钠肽能改善急性肺损伤病人Murray评分及氧合指数效果可能与调节NLRP3/6及白细胞介素水平有关.     

The inhibition of inducible nitric oxide synthase and oxidative stress by agmatine attenuates vascular dysfunction in rat acute endotoxemic model

Vascular dysfunction leading to hypotension is a major complication in patients with septic shock. Inducible nitric oxide synthase (iNOS) together with oxidative stress play an important role in development of vascular dysfunction in sepsis. Searching for an endogenous, safe and yet effective remedy was the chief goal for this study. The current study investigated the effect of agmatine (AGM), an endogenous metabolite of l-arginine, on sepsis-induced vascular dysfunction induced by lipopolysaccharides (LPS) in rats. AGM pretreatment (10 mg/kg, i.v.) 1 h before LPS (5 mg/kg, i.v.) prevented the LPS-induced mortality and elevations in serum creatine kinase-MB isoenzyme (CK-MB) activity, lactate dehydrogenase (LDH) activity, C-reactive protein (CRP) level and total nitrite/nitrate (NOx) level after 24 h from LPS injection. The elevation in aortic lipid peroxidation illustrated by increased malondialdehyde (MDA) content and the decrease in aortic glutathione (GSH) and superoxide dismutase (SOD) were also ameliorated by AGM. Additionally, AGM prevented LPS-induced elevation in mRNA expression of iNOS, while endothelial NOS (eNOS) mRNA was not affected. Furthermore AGM prevented the impaired aortic contraction to KCl and phenylephrine (PE) and endothelium-dependent relaxation to acetylcholine (ACh) without affecting endothelium-independent relaxation to sodium nitroprusside (SNP). In conclusion: AGM may represent a potential endogenous therapeutic candidate for sepsis-induced vascular dysfunction through its inhibiting effect on iNOS expression and oxidative stress.     

S-nitroso human serum albumin given after LPS challenge reduces acute lung injury and prolongs survival in a rat model of endotoxemia

Endotoxemia leads to the induction of inducible nitric oxide synthase (NOS-2) and increased expression of numerous inflammatory mediators contributing to endotoxin-induced acute lung injury. We tested the hypothesis that supplementation of nitric oxide (NO) by the novel NO donor S-nitroso human serum albumin (S-NO-HSA) given after lipopolysaccharide (LPS) challenge may reduce NOS-2 expression, lung inflammation and acute lung injury. Rats were divided into four groups: sham-operated (no treatment), LPS, LPS+HSA (human serum albumin), and LPS+S-NO-HSA. LPS was administered intravenously (20 mg kg(-1)) resulting in acute lung injury and a high mortality rate within 6 h (>90%). LPS-induced lung injury was characterized by an increased lung edema (lung wet/dry weight ratio), pulmonary neutrophil infiltration (myeloperoxidase activity, MPO) as well as a robust inflammatory response [increased expression of intercellular adhesion molecule-1 (ICAM-1), NOS-2, and cyclooxygenase-2 (COX-2)]. Infusion of S-NO-HSA or HSA was started 2 h after LPS and continued for 4 h (total dose of 72 mg kg(-1)) at a rate of 300 mug kg(-1) min(-1). S-NO-HSA but not HSA prolonged survival of endotoxemic rats, reduced the hypotensive response to LPS, minimized LPS-induced lung edema and injury, normalized MPO activity as well as diminished lung expression of pro-inflammatory molecules such as ICAM-1, NOS-2, and COX-2. Continuous supplementation of NO by S-NO-HSA after LPS challenge prevents induction of NOS-2, provides significant protection of endotoxin-induced acute lung injury, and prevents early mortality in endotoxic shock in rats. Our results suggest a potential therapeutic role for S-NO-HSA in endotoxemia.     

重组生长激素对小鼠血液细胞学指标变化的初步观察

目的 探讨重组生长激素（rhGH）对小鼠血液细胞学指标的变化。方法 应用CELL－DYN 1700及骨髓涂片来监测rhGH对30只小鼠的造血细胞增殖能力的作用。结果 阳性对照组周围血细胞计数明显低于生理盐水对照组（P＜0．01），实验组周围血细胞计数与生理盐水组比较无明显差异（P＞0．05）。实验组骨髓细胞形态学检查与生理盐水对照组比较，除粒细胞系统增生稍减低外，其它各系统均无明显变化。阳性对照组骨髓中有核细胞增生减低及明显减低，粒细胞系统形态异常。结论 rhGH对接受化疗小鼠的造血功能有一定的保护作用。     

Aggregation of recombinant human growth hormone induced by phenolic compounds

Phenolic compounds can be used as antimicrobial preservative agents in pharmaceutical formulations. Unfortunately, these compounds often adversely affect proteins, triggering aggregation in particular. In this study, a variety of phenolic compounds and structurally similar non-aromatic alcohols were investigated for their role in causing the aggregation of a model therapeutic protein, recombinant human growth hormone (rhGH). As determined by various methodologies, most of the phenolic compounds caused rhGH aggregation, especially at high concentrations. Stress studies under freezing, high-temperature incubation, and agitation suggest that the destabilizing influence of the compounds tested increases in the order of benzyl alcohol < phenol × resorcinol < catechol < meta-cresol < 2-chlorophenol. Non-aromatic alcohols, except 2,6-dimethylcyclohexanol, have a much less adverse effect. Determination of the thermal transition temperature by microcalorimetry studies also reflected this trend. From our study, we conclude: (1) the phenolic additive-induced rhGH aggregates were held by non-covalent forces; (2) no significant physical binding occurred between the protein and these compounds; (3) the aggregation tendency of the phenolic compounds failed to correlate with their hydrogen bonding strength; (4) the presence of a phenolic additive caused conformational changes in rhGH's structure; (5) the effect of these phenolic compounds on rhGH aggregation decreased at high and low pHs.