从气、血、水“三位一体”论治糖尿病肾脏病探析

糖尿病肾脏病的主要病位在于肾,涉及肺、脾、肝、心、膀胱、三焦等脏腑,病性为本虚标实,在其发生、发展过程中,常常"气""血""水"三者同时发生病变,其中"气"的病变有气虚、气滞、气逆、气虚及阳四层含义,"血"的病变有阴虚、血虚、血瘀三层含义,"水"的病变有水湿、水饮、湿浊、湿热四层含义。三者相互影响,互为因果,且在不同的发展阶段病变层次不一,因此,在治疗应该在分期辨证的基础上,重视气、血、水同治,早期应重视益气养阴,中晚期重视滋阴温阳,活血化瘀利湿贯穿始终。     

靶向特异基因小干扰RNA预防乙醇性股骨头坏死的研究

目的 观察靶向PPARγ基因小干扰RNA(siRNA)预防灌胃法家兔乙醇性股骨头坏死的效果.方法 将96只家兔随机分为4组,每组24只.N组:灌胃法给予生理盐水10ml/(kg·d).M组:灌胃法给予烈性酒(含乙醇度46%,V/V)10 ml/(kg·d);S组:实验1、3、5个月第1天,随机选择侧别手术,穿刺注入股骨头内25 μl siRNA重组腺病毒,同M组灌酒;CO组:同S组手术,同M组灌酒,不注入siRNA重组腺病毒.于2、4、6个月末分批处死家兔,观察血清学和股骨头组织病理学变化.结果　实验6个月时,N、M、CO、S组血清甘油三酯含量分别为(1.21±0.12)、(4.59±1.58)、(4.63±1.17)、(4.32±1.20)mmol/L;总胆固醇含量分别为(2.35±0.33)、(19.59±1.58)、(20.13±1.17)、(18.32±1.20)mmol/L;脂肪细胞平均直径分别为(40.89±2.41)、(48.65±2.93)、(49.45±2.63)、(42.52±2.57)μm;骨小梁面积分数分别为(41.80±2.47)%、(30.70±2.86)%、(29.80±2.69)%、(41.60±2.87)%;空骨陷窝计数分别为(12.30±1.73)%、(22.40±1.52)%、(23.10±1.62)%、(11.70±1.46)%;PPARγ表达量分别为(0.29±0.05)、(0.66±0.12)、(0.61±0.15)、(0.33±0.05);骨钙素mRNA表达量分别为(0.92±0.07)、(0.19±0.11)、(0.23±0.08)、(0.88±0.11);PPARγ蛋白表达量分别为(0.75±0.08)、(1.60±0.11)、(1.55±0.12)、(0.65±0.05).M、CO组病理变化明显,髓内造血组织减少,脂肪细胞增殖肥大,骨小梁变细、稀疏、部分断裂;S组股骨头内最大脂肪细胞平均直径、空骨陷窝计数、骨坏死发生率、PPARγ基因与蛋白表达量均明显低于M、CO组(P＜0.01,P＜0.05),S组与N组差异无统计学意义(P＞0.05).结论　靶向PPARγ基因siRNA腺病毒载体能够有效阻断乙醇诱导的家兔股骨头内MSCs中PPARγ基因表达及成脂分化,预防乙醇性ONFH的发生.

Abstract：

Objective To observe the effect of small interfering RNA (siRNA) adenovirus vectors targeting PPARγto prevent the alcohol-induced osteonecrosis of the femoral head (ONFH) in rabbits.Methods Ninety-six rabbits were randomly divided into four groups. In group N, normal saline ( 10 ml/kg every day) was poured into stomach. In group M, the strong wine (the volume fraction was 46% alcohol,10 ml/kg every day) was poured into stomach. In group S, under the randomly selected side and anesthesia, the animals were injected with 25 μl siRNA adenovirus drip into the femoral head and then the puncture hole was closed on the 1st day at the 1st, 3rd and 5th month of the experiment, and at the same time the strong wine (the volume fraction was 46% alcohol, 10 ml/kg every day) was poured into stomach. In group CO, animals were treated with the same method as the group S, but not injected with siRNA adenovirus drip. The animals were sacrificed in batches at 2nd, 4th and 6th month after the experiment. The serology and pathological changes of the femoral head were studied. Results At the 6th month, the triglyceride (TG, mmol/L) contents in groups N, M, CO and S were (1.21 ±0. 12), (4.59 ± 1.58), (4.63 ±1.17) and (4. 32 ± 1.20), the cholesterol levels ( CHO, mmol/L) were (2. 35 ± 0. 33), ( 19. 59 ±1.58), (20. 13 ± 1. 17) and ( 18.32 ± 1.20), the average diameter of the max adipocyte (μm) was (40. 89 ± 2. 41 ), (48.65 ± 2.93 ), (49. 45 ± 2. 63 ) and ( 42. 52 ± 2. 57 ), the trabeculace area fraction was (41.80 ±2. 47)%, (30. 70 ±2. 86)%, (29. 80 ±2. 69)% and (41.60 ±2. 87)%, the percentage of empty osteocyte lacunae was (12.30 ± 1.73)%, (22.40 ± 1.52)%, (23. 10 ± 1.62)% and (11.70 ±1.46)%, the expreasion levels of PPARγ mRNA were (0.29 ±0.05), (0.66±0. 12), (0.61±0.15) and (0. 33 ± 0. 05 ), the expression levels of osteocalcin mRNA were (0. 92 ± 0. 07 ), (0. 19 ± 0. 11 ), (0. 23 ±0. 08) and (0. 88 ± 0. 11 ), the expression levels of PPARγprotein were ( 0. 75 ± 0. 08 ), ( 1.60 ± 0. 11 ),(1.55 ±0. 12) and (0.65 ±0.05), respectively. In groups M and CO, the pathological changes were obvious; there was decreased hematopoietic tissue, proliferation and hypertrophy of the adipocytes, increased fatty tissue, and thinned, sparse or breoken bone trabeculae in the femoral head; the area fraction of the trabeculae was reduced. In group S, the average diameter of the max adipocyte, percentage of empty osteocyte lacunae, incidence of ONFH, the expressions of PPARγmRNA and protein were obviously reduced as compared with groups M and CO (P ＜0. 01,P ＜0. 05). There was no statistically significant difference between group S and group N (P ＞ 0. 05). Conclusion siRNA adenovirus vectors targeting PPARγcan efficaciously suppress the expression of PPARγ gene and adipogenic differentiation of the MSCs in the femoral head induced by alcohol, which may prevent the development of the alcohol-induced ONFH in rabbits.     

骨髓干细胞和伞状支撑骨移植治疗股骨头坏死的动物实验研究报告

目的 探讨骨髓干细胞移植和伞状支撑骨移植治疗股骨头坏死的效果.方法 取6只健康山羊,采用液氮冷冻法制作单侧股骨头坏死动物模型,8周后在坏死侧行骨髓干细胞移植和伞状支撑骨移植术.分别于术后3、6个月做放射学检查,观察股骨头内骨质变化.结果 实验动物一般状况良好.液氮冷冻法8周后达成股骨头坏死模型,模型侧后肢出现跛行.骨髓干细胞移植和伞状支撑骨移植术后3个月X线显示,股骨头外形恢复,股骨头内囊性低密度区消失,可见骨柱影,股骨头内骨质愈合状况良好,原塌陷已修复,股骨头无再塌陷.6个月时,治疗侧股骨头外形正常,骨密度基本均匀,骨柱影已模糊,植骨已融合,股骨头无再塌陷.正常对照侧股骨头无异常变化.结论 骨髓干细胞移植和伞状支撑骨移植术可以有效治疗骨坏死,防治股骨头塌陷,效果良好. Abstract： Objective To study the usefulness to treat the osteonecrosis of the femoral head (ONFH) with bone marrow mesenchymal stem cell transplantation and umbrella strut bone grafting. Methods Six goats were established of osteonecrosis of the femoral head models on one side by method of liquid nitrogen frozen, and then they were taken into bone marrow mesenchymal stem cell transplantation and umbrella strut bone grafting after 8 weeks. The radiological examination was made at 3 and 6 months after the bong grafting, and observed changes of bone union in the femoral head. Results The general state of the experimental animal was fine after the operation. The model of ONFH was reached on 8 weeks after liquid nitrogen frozen, and limping. After 3 months of bone marrow mesenchymal stem cell transplantation and umbrella strut bone grafting, X-ray film showed that low bone density disappeared, the shape of the femoral heads, grafting strut bone and initial bone union were fine in the head, and no repeated collapse of the head. The X-ray film showed that the shape of the femoral heads was normal, grafting strut bone was union in the head, and no repeated collapse of the head after 6 months. There was normal on the control side. Conclusions The ONFH can be treated effectively by bone marrow mesenchymal stem cell transplantation and umbrella strut bone grafting, and the collapse of the femoral head is prevented.     

吕仁和教授治疗慢性肾脏病经验

吕仁和教授长期从事慢性肾脏病的诊疗工作,积累了丰富的经验,他总结的"六对论治"方法在临床上诊治慢性肾脏病取得了较好疗效。从吕仁和教授"六对论治"中对病论治、对症论治、对症辨证论治、对症辨病与辨证论治相结合4个方向来介绍吕仁和教授治疗慢性肾脏病的经验。     

糖尿病肾脏病中医治法研究进展

糖尿病肾脏病是糖尿病微血管并发症之一,已成为导致终末期肾衰的首要原因,中医治疗DKD有几千年的历史,有丰富的理论体系和许多临床实用方药。中医治法研究是糖尿病肾脏病研究的热点之一,观今之临床,大致有补肾化瘀治法、补虚活血泻浊治法、益气活血治法、化瘀散结治法、从热论治、从络论治、从风论治,深入挖掘其发展渊源、理论内涵对于提高糖尿病肾脏病的临床疗效具有重要意义。     

吕仁和治疗顽固性头痛案2则

介绍吕仁和教授治疗顽固性头痛验案2则,并总结了吕教授治疗顽固性头痛的辨治思路和用药特色。吕仁和教授主张从五脏论治顽固性头痛,尤为肝脏,认为其病理因素多与痰、瘀相关,治则为调和五脏、燮理阴阳、气血同调,治法重视平抑肝阳、滋阴潜阳、行气活血、化痰通络,对于情绪相关顽固性性头痛多用酸枣仁、百合、玫瑰花、龟甲为主方进行加减;四逆散临床应用不可囿于疏肝理气,功在行气活血。     

山羊骨髓间充质干细胞的体外培养与生物学特性

目的:体外分离培养山羊骨髓间充质于细胞(BMSC),并观察其生物学特性.方法:应用常规密度梯度离心法从山羊骨髓中分离BMSC并传代培养,镜下观察细胞形态及生长特性,绘制细胞生长曲线,测定倍增时间,免疫细胞化学法鉴定其表型.结果:原代培养的BMSC初始呈圆形、梭形及多角形等,细胞呈贴壁生长,48 h可见贴壁细胞有伸展现象,细胞渐变为均一梭形,14 d时可达90%融合.第1、3和5代BMSC生长曲线呈"S"型,1～2d为潜伏期,2～3 d后进入对数增长期.7～8 d后进入平台期,平均倍增时间为(23.2±1.9)h.BMSC CD29和CD44呈广泛阳性表达,CD34和CD45呈阴性表达.结论:成功分离培养出山羊BMSC,细胞纯度较高,生长状态良好.     

益气祛风通络方对糖尿病肾病大鼠尿蛋白的影响

目的通过动物实验,以西药厄贝沙坦为对照,观察中药益气祛风通络方对糖尿病肾病大鼠尿蛋白、血糖、肾功能的影响,探索益气祛风通络方治疗糖尿病肾病的疗效。方法将35只Wister大鼠随机分成正常组、假手术组、模型组、中药组、西药组,通过高脂高热量喂养加腹腔注射链脲佐菌素加单侧肾切除术复制糖尿病肾病模型,检测各组大鼠24小时尿蛋白,尾尖取血测血糖,腹主动脉取血测血清肌酐。结果 (1)干预前后,各组的24小时尿蛋白差值分别是中药组(-10.80±5.02)×10~(-3 )g、西药组(-3.38±8.67)×10~(-3 )g、模型组(15.48±17.95)×10~(-3)g、假手术组(1.22±2.19)×10~(-3 )g、正常组(-0.16±1.48)×10~(-3 )g,中药组大鼠24小时尿蛋白定量明显下降,与假手术组相比P=0.007,与正常组相比P=0.015,差异都有统计学意义(P0.05)。(2)在药物干预的12周里,只有中药组的血糖在持续下降,其余各组血糖值上升,中药组干预前后血糖差值与西药组相比P=0.001,与模型组相比P=0.000,与假手术组相比P=0.001,与正常组相比P=0.000,差异均有统计学意义(P0.05)。(3)干预12周后,各组大鼠腹主动脉取血,测血清肌酐值分别是中药组(29.23±1.65)μmol/L、西药组(41.52±5.47)μmol/L、模型组(53.33±12.76)μmol/L、假手术组(31.42±2.61)μmol/L、正常组(29.18±1.37)μmol/L,中药组血肌酐值较低,与西药组相比P=0.011,与模型组相比P=0.028,差异均有统计学意义(P0.05);模型组血肌酐值较高,与假手术组相比P=0.040,与正常组相比P=0.028,差异均有统计学意义(P0.05)。结论益气祛风通络方对糖尿病肾病大鼠的24小时尿蛋白有降低作用,同时还能降低血糖、保护肾功能,疗效优于西药厄贝沙坦。