荷负电气溶胶治疗对大鼠烫伤创面白细胞介素-8、10表达的影响及意义

Objective To discuss the influence of aerosol bioelectricity on the expression of interleukin (IL) -8 and IL-10 in wound healing of burned rats. Methods The deep Ⅱ degree scalding models were established in Sprague Dawley (SD) rats. Rats were randomly divided into experimental group (n1 =20) and control group (n2 =20). The rats in experimental group were treated with aerosol bioelectricity.Samples were collected at the first to eleventh day post-scalding. Immunohistochemistry and image analysis methods were conducted to examine the expression of IL-8 and IL-10 in both experimental and control groups. Results The average wound healing time in experimental group was 7. 00 ± 1. 15 days, and that in control group was 9. 00 ± 1. 34 days. IL-8 and IL-10 were observed mainly in polylmorphonuclear and mononuclear cells in both experimental and control groups on the 1 st day. On the third day, fibroblasts abounded, IL-8 expression was increased evidently and reached a peak. The peak value (6. 73 ± 1. 36) in experimental group was lower significantly than that in control group ( 2. 85 ± 0. 72, P ＜ 0. 01). From the 5th to 11th day, IL-8 expression was declined rapidly. IL-10 was expressed in keratode cells and had the peak value in experimental group (1. 24 ±0. 15) and control group (5. 69 ± 1. 32) on the 3rd day. IL-10 expression was declined gradually from the 5th to 11th days. The expression level of IL-10 in experimental group was significantly higher than in control group from the 3rd day to 11th days post-scalding (P＜0. 01). On the 3rd day, both IL-8 and IL-10 in experimental and control groups were expressed abundantly , and there was negative relationship between them (r = - 0. 862, P ＜ 0. 01). Conclusion Aerosol bioelectricity can indicate active cells proliferation through down-regulating the expression of IL-8 and up-regulating the expression of IL-10, accelerating burned wound healing.     

荷负电气溶胶治疗对大鼠烫伤创面白细胞介素-8、10表达的影响及意义

Objective To discuss the influence of aerosol bioelectricity on the expression of interleukin (IL) -8 and IL-10 in wound healing of burned rats. Methods The deep Ⅱ degree scalding models were established in Sprague Dawley (SD) rats. Rats were randomly divided into experimental group (n1 =20) and control group (n2 =20). The rats in experimental group were treated with aerosol bioelectricity.Samples were collected at the first to eleventh day post-scalding. Immunohistochemistry and image analysis methods were conducted to examine the expression of IL-8 and IL-10 in both experimental and control groups. Results The average wound healing time in experimental group was 7. 00 ± 1. 15 days, and that in control group was 9. 00 ± 1. 34 days. IL-8 and IL-10 were observed mainly in polylmorphonuclear and mononuclear cells in both experimental and control groups on the 1 st day. On the third day, fibroblasts abounded, IL-8 expression was increased evidently and reached a peak. The peak value (6. 73 ± 1. 36) in experimental group was lower significantly than that in control group ( 2. 85 ± 0. 72, P ＜ 0. 01). From the 5th to 11th day, IL-8 expression was declined rapidly. IL-10 was expressed in keratode cells and had the peak value in experimental group (1. 24 ±0. 15) and control group (5. 69 ± 1. 32) on the 3rd day. IL-10 expression was declined gradually from the 5th to 11th days. The expression level of IL-10 in experimental group was significantly higher than in control group from the 3rd day to 11th days post-scalding (P＜0. 01). On the 3rd day, both IL-8 and IL-10 in experimental and control groups were expressed abundantly , and there was negative relationship between them (r = - 0. 862, P ＜ 0. 01). Conclusion Aerosol bioelectricity can indicate active cells proliferation through down-regulating the expression of IL-8 and up-regulating the expression of IL-10, accelerating burned wound healing.     

荷负电气溶胶治疗对大鼠烫伤创面白细胞介素-8、10表达的影响及意义

目的 观察荷负电气溶胶治疗对大鼠烫伤创面愈合过程中白细胞介素(IL)-8、IL-10表达的影响,探讨荷负电气溶胶治疗促进创面愈合的作用机制.方法 制作SD大鼠深Ⅱ度烫伤模型,采用分组对照方法,将40只鼠随机分为治疗组(n1=20)和对照组(n2=20).治疗组应用荷负电气溶胶治疗,每次1.5 h,每天2次,直至创面愈合;对照组不作荷负电气溶胶治疗.伤后第1～11天分别取创面标本制作切片,采用免疫组织化学和图像分析方法,检测创面愈合过程中IL-8和IL-10表达水平.结果 创面平均愈合时间治疗组为(7.00±1.15)d,对照组为(9.00±1.34)d,治疗组创面愈合时间明显提前(P＜0.01).免疫组织化学显示,两组IL-8均在伤后第1天开始表达.主要位于多核粒细胞和单核细胞;第3天表达明显增多达高峰,并见大量成纤维细胞表达,治疗组的峰值明显低于对照组,差异有统计学意义(P＜0.01),第5～11天表达水平迅速下降.两组IL-10伤后第1天在淋巴细胞和单核细胞均有表达;第3天开始有角质细胞表达并达高峰,第5～11天表达水平缓慢下降,但治疗组要明显高于对照组,第3～11天差异有统计学意义(P＜0.01).结论 荷负电气溶胶治疗能有效抑制创面IL-8的表达及促进IL-10的表达,缩短炎症进程,从而加速创面愈合.

Abstract：

Objective To discuss the influence of aerosol bioelectricity on the expression of interleukin (IL) -8 and IL-10 in wound healing of burned rats. Methods The deep Ⅱ degree scalding models were established in Sprague Dawley (SD) rats. Rats were randomly divided into experimental group (n1 =20) and control group (n2 =20). The rats in experimental group were treated with aerosol bioelectricity.Samples were collected at the first to eleventh day post-scalding. Immunohistochemistry and image analysis methods were conducted to examine the expression of IL-8 and IL-10 in both experimental and control groups. Results The average wound healing time in experimental group was 7. 00 ± 1. 15 days, and that in control group was 9. 00 ± 1. 34 days. IL-8 and IL-10 were observed mainly in polylmorphonuclear and mononuclear cells in both experimental and control groups on the 1 st day. On the third day, fibroblasts abounded, IL-8 expression was increased evidently and reached a peak. The peak value (6. 73 ± 1. 36) in experimental group was lower significantly than that in control group ( 2. 85 ± 0. 72, P ＜ 0. 01). From the 5th to 11th day, IL-8 expression was declined rapidly. IL-10 was expressed in keratode cells and had the peak value in experimental group (1. 24 ±0. 15) and control group (5. 69 ± 1. 32) on the 3rd day. IL-10 expression was declined gradually from the 5th to 11th days. The expression level of IL-10 in experimental group was significantly higher than in control group from the 3rd day to 11th days post-scalding (P＜0. 01). On the 3rd day, both IL-8 and IL-10 in experimental and control groups were expressed abundantly , and there was negative relationship between them (r = - 0. 862, P ＜ 0. 01). Conclusion Aerosol bioelectricity can indicate active cells proliferation through down-regulating the expression of IL-8 and up-regulating the expression of IL-10, accelerating burned wound healing.     

Effect of substance P released from peripheral nerve ending on endogenous expression of epidermal growth factor and its receptor in wound healing

Objective: To explore the relationship between substance P (SP) released from peripheral nerve endings and the expression of epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) during wound bealing. Methods: Fifty Wistar rats were randomly divided into 2 groups, injury group and capsaicin group. In the injury group, a full-thickness skin wound on the back of the rat was taken. The wound edge and granulation tissues were taken on the 1st, 3rd, 6th, 9th, 12th days after injury, respectively. In the capsaicin group, capsaicin was injected subcutaneously on the back of the rats to destroy the sensory nerve to prevent the secretion of SP, then a wound and sample was made in the same way.Immunohistochemistry and in situ hybridization were employed to detect the expression of SP, EGF/EGFR, and EGF mRNA/EGFR mRNA in the granulation tissues.Results: In the injury group,immunohistochemical stain of SP and EGF/EGFR was located on the hair follicles and sebaceous glands at the 1st day. And the stain of SP was obvious at the 3rd day in the granulation tissues, then decreased gradually. EGF/EGFR was at low level at the 3rd day, then increased gradually and reached the peak at the 9th day, then declined. In the capsaicin group, the stain of SP and EGF/EGFR was faint and without obvious change during the wound healing process. The tendency of the EGF mRNA/EGFR mRNA expression was similar to that of EGF/EGFR. Conclusions: During wound healing, SP may promote the healing process by affecting the expression of EGF/EGFR in the erunuation tissues.     

雌激素对大鼠创面愈合过程中微血管形成的影响及其机制

目的 观察雌激素对大鼠全层皮肤缺损创面愈合过程中微小血管再形成的影响.方法 将40只雌性Wistar大鼠随机分成3组:单纯损伤组15只(A组),去卵巢组15只(B组)和假手术组10只(C组).8周后,在背部制成全层全损皮肤创伤愈合模型,分别于伤后1、3、7、14和21 d记录3组各个时间点伤腔容积,并在创缘取材.常规组织学观察皮肤愈合过程和微血管的数量,同时采用雌激素β受体(ER-β)进行免疫组织化学染色,观察微血管及周围细胞受体的染色.结果　比较各个时间点的伤腔容积,单纯损伤组和假手术组大鼠创面内肉芽组织形成量明显大于去卵巢组,组织学评分略高于去卵巢大鼠;而单纯损伤组和假手术组之间差异无统计学意义(P＞0.05).卵巢切除大鼠伤后各时间点,真皮内微小血管的数量较单纯损伤组减少,各组均于伤后7 d,随着成纤维细胞和毛细血管数目的 增加,肉芽组织生长达到高峰.同时,各损伤组创基内微小血管周围均出现较为幼稚的细胞,其表面可以表达ER-β,但卵巢切除组(159.91±12.65)稍弱于单纯损伤组(189.36±27.32),差异无统计学意义(P＞0.05).结论　大鼠去卵巢后创面愈合速度明显减慢与微血管形成有关,雌激素及其受体在该过程中有重要作用.

Abstract：

Objective To investigate the effect of estrogen on the revascularization of full-thickness skin wounds healing in rats. Methods Forty 2-month-old female Wistar rats were divided into three groups: control group (group A, pure injury,n = 15), ovariectomized group (group B,n = 15) and sham operation group (group C,n = 10 ). After 8 weeks, full-thickness wounds model, 18 mm in diameter, were made on the back of the each animal. The volume of wound was measured at each time point. The specimens of wound edge were obtained at the 1st, 3rd, 7th, 14th and 21st day after injury. The process of wound healing and the number of microvessels in the granulation tissue were evaluated by Haematine-Eosin (HE) staining. The immunohistochemical staining was used to detect the expression of estrogen receptor β (ER-β) in microvascular endothelial cells and their surrounding cells. Results To compare the volume of wound at each time point, the granulation tissue formation in group A and group C was significantly higher than that in group B, but there was no significant difference between group A and group C. The number of capillary blood vessels in group B was significantly reduced as compared with that in group A. The expression of ER-β was positive on the cells surface in all groups. ER-β expression in group B was weaker than in group A ( 159. 91 ± 12. 65 v. s 189. 36 ±27. 32,P ＜0. 05). Conclusion The delay of wound healing in ovariectomized rats has a great relationship with angiogenesis. Estrogen and its receptor might play important roles in the process.     

IGFBPrP1对小鼠肝组织胶原含量的影响及其机制的研究

目的 明确外源性胰岛素样生长因子结合蛋白相关蛋白1(IGFBPrP1)对肝组织胶原含量的影响及可能的机制.方法 24只雄性C57BL/6野生型小鼠随机分为3组:正常对照组、rmIGFBPrP1 2周组和rmIGFBPrP1 4周组,每组8只.取肝组织行HE、苦味酸-天狼猩红、免疫组织化学染色及Western blot检测.结果 HE和苦味酸-天狼猩红染色显示外源性rmIGFBPrP1可使肝组织发生病理改变,并导致肝组织中胶原含量显著增多(P＜0.05).Western blot检测显示rmIGFB-PrP1 2周组及4周组肝组织中IGFBPrP1、纤维连接蛋白(FN)、Ⅰ型胶原(Collagen Ⅰ)的表达明显增强,且4周组表达量较2周组高(P＜0.01).免疫组织化学染色显示Ⅲ型胶原(Collagen Ⅲ)在rmIG-FBPrP1 2周组及4周组表达量明显增高,且4周组高于2周组(P＜0.01).Western blot及免疫组织化学染色共同显示转化生长因子β1(TGF-β1)、Smad3、p-Smad2/3在rmIGFBPrP1 2周组及4周组表达显著增强,且4周组强于2周组(P＜0.01).结论 外源性IGFBPrP1可通过TGF-β1/Smad3信号通路导致肝组织中胶原含量明显增加、ECM过度沉积.

Abstract：

Objective To investigate the effect and mechanism of exogenous IGFBPrP1 on collagen content in liver tissue. Methods Twenty-four male C57BL/6 wild-type mice were randomly divided into three groups: Control group (n=8), rmIGFBPrP1 2 weeks group (n=8) and rmIGFBPrP1 4weeks group (n=8). Both hematoxylin-eosin (HE) staining and picric acid-Sirius red staining were performed. The protein expression of IGFBPrP1, Collagen Ⅰ , Collagen Ⅲ, FN, TGF-β1, Smad3 andp-Smad2/3 was evaluated by immunohistochemistry or Western blot. Results Exogenous IGFBPrP1 can cause pathological changes in liver tissue. Collagen content was significantly increased by hematoxylin-eosin (HE) staining and picric acid-Sirius red staining (P＜0.05). The protein expression of IGFBPrP1, FN and Collagen Ⅰ was gradually increased after rmIGFBPrP1 injection for 2 weeks and 4 weeks by Western blot (P＜0.01). The protein expression of Collagen Ⅲ was obviously increased in the rmIGFBPrP1 2 weeks group and rmIGFBPrP1 4 weeks group by immunohistochemistry, and the level in the 4 weeks group was higher than that in the 2 weeks group (P＜0. 01). The protein expression of TGF-β1, Smad3 and p-Smad2/3 in liver tissue was significantly increased after rmIGFBPrP1 injection in a time-dependent manner by both immunohistochemistry and Western blot (P＜0. 01).Conclusion Exogenous IGFBPrP1 can cause a marked increase in collagen content and the excessive deposition of ECM through the TGF-β1/Smad3 pathway in liver tissue.     

不同来源的表皮干细胞促进皮肤创面愈合的研究

目的 观察表皮干细胞(ESCs)及去分化来源的表皮干细胞(DESCs)对皮肤创面愈合的促进作用及其异同.方法 由新鲜人包皮标本以贴壁法分离ESCs,随机分为3组,第1组作为原代ESCs:第2组传代培养至第6代时可见细胞成熟呈表皮细胞(ECs)形态,由原来的大克隆生长的小圆细胞形态变为较为肥大的不规则型细胞,无法形成克隆样增殖;其细胞表型也由β1整合素、CK19强阳性、CK10阴性变为β1整合素、CK19阴性,而CK10强阳性,予以100μg/L的碱性成纤维细胞生长因子(bFGF)培养48 h进行去分化诱导,48 h后ECs周边开始出现小圆细胞并呈克隆样生长,β1整合素、CK19、CK10等细胞表型也趋于与ESCs一致,即DESCs;第3组传代培养至第6代获得ECs.3组细胞皆以D-Hanks液重悬调其终质量浓度为2×106个/ml.将12只裸鼠每只背部左右各制备1个直径6 mm创面,共24个创面.将全部创面随机等分为4组,每组6个,分别以ESCs、DESCs、Ecs及生理盐水(NS)各0.2 ml进行创面注射.于术后0、3、7 d测量创面面积进行统计学分析,并于术后7 d行创面取材苏木素-伊红(HE)染色.结果 ESCs组与DESCs组在术后3 d时创面面积分别为(11.758±2.544)、(11.515±1.351)mm2,7 d时创面面积分别为(1.795±1.063)、(2.043±1.138)mm2,修复速度要明显快于ECs组与NS组[3 d时创面面积分别为(17.857±1.722)、(16.192±2.256)mm2,7 d时创面面积分别为(5.367±1.219)、(5.070±1.357)mm2,P＜0.01];而ESCs与DESCs组之间创面修复速度比较差异无统计学意义(P＞0.05);ECs组与NS组之间比较差异无统计学意义(P＞0.05).HE染色提示ESCs组与DESCs组的创面愈合质量优于ECs组与NS组,可见有皮肤附属腺的再生且纤维瘢痕组织较少.结论 ESCs与DESCs皆有促进创面愈合的作用,相互之间无明显差异.

Abstract：

Objective To investigate the enhancing effect of epithelial stem cells (ESCs) and dedifferentiation derived epithelial stem cells (DESCs) in the healing of epidermal wound. Methods ESCs were isolated from the fresh circumcised foreskins by using adherence method and randomly divided into three cohorts: ESCs cohort, DESCs cohort and epithelial cells (ECs) cohort. The final cell density was adjusted to 2 × 106/ml with D-Hanks in each group. The model of BALB/C mice full-thickness skin loss wounds was established. Two circular 6 mm-diameter skin loss wounds were cut on every BALB/C mouse back. Twenty-four wounds of 12 BALB/C mice were divided equally and randomly into 4 groups:DESCs, ESCs, ECs and NS. The cell suspensions of DESCs, ESCs and ECs were injected respectively into the wound edges with the density of 2 × 106/ml. And the volme was 0. 2 ml per treatment wound. The same volume of normal saline was injected in NS group. On the postoperative day 0, 3, 7, all of the wound areas were measured and analyzed statistically. On the postoperative 7 day, the sections were made and observed by using hematoxylin and eosin (HE) staining. Results The wound areas in DESCs group and ESCs group were ( 11. 758 ± 2. 544) and ( 11.515 ± 1.351 ) mm2 on the 3rd day postoperatively, and ( 1. 795 ±1. 063) and (2. 043 ± 1. 138) mm2 on the 7th day postoperatively, respectively. The wound areas in ECs group and NS group were ( 17. 857 ± 1. 722) and ( 16. 192 ±2. 256) mm2 on the 3rd day postoperatively,and ( 1. 795 ± 1. 063) and (2. 043 ± 1. 138) mm2 on the 7th day postoperatively, respectively. There was statistically significant difference in wound area between ESCs or DESCs group and ECs, NS groups on the 3rd, 7th day postoperatively ( P ＜ 0. 01 ), but there was no statistically significant difference between DESCs and ESCs groups ( P ＞ 0. 05 ). HE staining showed the repairing quality of treatment wounds was superior to that in the control wounds, and regenerating glands and less fibrous connective tissues were seen in ESCs and EDSCs groups. Conclusion Both ESCs and DESCs could promote the wound repair.     

低蛋白饮食对环孢素A肾病大鼠肾间质纤维化的作用

Objective To investigate whether low-protein diet has protective effect on the progression of renal interstitial fibrosis in rats with cyclosporine A (CsA)-induced nephropathy. Methods Eighteen male Sprague-Dawley rats were randomly divided into three groups (6 rats in each group). The rats in control group (C group) received common diet; in model group (M group) low-salt diet; in intervention group (Ⅰ group) low-salt and low-protein diet. After diet adaptation period of one week, the rats in C group received subcutaneous injection of olive oil 1 mg/kg daily for 5 weeks, while M group and Ⅰ group subcutaneous injection of CsA (diluted into 25 g/L with olive oil) 1 ml/kg for 5 weeks. All the rats were sacrificed at the end of the 5th week. The food-intake and body weight were measured daily. The creatinine clearance (Ccr) was examined before rats were sacrificed. The semi-quantitative pathological analysis on kidney sections was performed. The mRNA and protein expression of transforming growth factor-β1 (TGF-βI) and type Ⅰ collagen (Col Ⅰ) in kidney tissue was determined with real time PCR and immunohistochemical staining, respectively. Results The food-intake and body weight of rats in M and I groups were significantly lower than those in C group (P<0.05). Compared with C group, the Ccr levels in M and Ⅰ groups were significantly reduced [(0.65±0.15) ml/min, (0.40+0.13) ml/min vs (1.55±0.29) ml/min, P<0.05], the relative fibrosis areas of kidney interstitium in M and I groups were significantly increased (3.60%±0.46%, 3.26%±0.75% vs 0.44%±0.24%, P<0.05), the mRNA and protein expression of TGF-β1 in M and I group was significantly up-regulated (by 2.6 and 3.1 times in mRNA and by 1.5 and 1.6 times in protein, respectively, P<0.05), and the mRNA and protein expression of Col Ⅰ in M and I groups was also significantly up-regulated (by 3.0 and 3.5 times in mRNA and by 2.3 and 2.1 times in protein, respectively, P<0.05). There were no significant differences between M and I groups in every parameters above-mentioned except the rat body weight and Ccr. Both the body weight and Ccr in Ⅰ group were significantly lower than those in M group (P<0.05). Compared with C group, the urine osmotic pressure in M group and in I group were deceased (for M group, P>0.05; for I group, P<0.05). Compared with C group, the serum cholesterol levels in M and I groups were significantly increased (P<0.05), and the serum phosphorus level in I group was significantly decreased (P<0.05). The levels of serum albumin and serum calcium of all three groups had no statistical differences (P>0.05). Conclusion Low-protein diet has no renoprutective effects on the rat model of cyclosporin A nephropathy, on the contrary, may induce body weight loss.     

低蛋白饮食对环孢素A肾病大鼠肾间质纤维化的作用

Objective To investigate whether low-protein diet has protective effect on the progression of renal interstitial fibrosis in rats with cyclosporine A (CsA)-induced nephropathy. Methods Eighteen male Sprague-Dawley rats were randomly divided into three groups (6 rats in each group). The rats in control group (C group) received common diet; in model group (M group) low-salt diet; in intervention group (Ⅰ group) low-salt and low-protein diet. After diet adaptation period of one week, the rats in C group received subcutaneous injection of olive oil 1 mg/kg daily for 5 weeks, while M group and Ⅰ group subcutaneous injection of CsA (diluted into 25 g/L with olive oil) 1 ml/kg for 5 weeks. All the rats were sacrificed at the end of the 5th week. The food-intake and body weight were measured daily. The creatinine clearance (Ccr) was examined before rats were sacrificed. The semi-quantitative pathological analysis on kidney sections was performed. The mRNA and protein expression of transforming growth factor-β1 (TGF-βI) and type Ⅰ collagen (Col Ⅰ) in kidney tissue was determined with real time PCR and immunohistochemical staining, respectively. Results The food-intake and body weight of rats in M and I groups were significantly lower than those in C group (P<0.05). Compared with C group, the Ccr levels in M and Ⅰ groups were significantly reduced [(0.65±0.15) ml/min, (0.40+0.13) ml/min vs (1.55±0.29) ml/min, P<0.05], the relative fibrosis areas of kidney interstitium in M and I groups were significantly increased (3.60%±0.46%, 3.26%±0.75% vs 0.44%±0.24%, P<0.05), the mRNA and protein expression of TGF-β1 in M and I group was significantly up-regulated (by 2.6 and 3.1 times in mRNA and by 1.5 and 1.6 times in protein, respectively, P<0.05), and the mRNA and protein expression of Col Ⅰ in M and I groups was also significantly up-regulated (by 3.0 and 3.5 times in mRNA and by 2.3 and 2.1 times in protein, respectively, P<0.05). There were no significant differences between M and I groups in every parameters above-mentioned except the rat body weight and Ccr. Both the body weight and Ccr in Ⅰ group were significantly lower than those in M group (P<0.05). Compared with C group, the urine osmotic pressure in M group and in I group were deceased (for M group, P>0.05; for I group, P<0.05). Compared with C group, the serum cholesterol levels in M and I groups were significantly increased (P<0.05), and the serum phosphorus level in I group was significantly decreased (P<0.05). The levels of serum albumin and serum calcium of all three groups had no statistical differences (P>0.05). Conclusion Low-protein diet has no renoprutective effects on the rat model of cyclosporin A nephropathy, on the contrary, may induce body weight loss.     

低蛋白饮食对环孢素A肾病大鼠肾间质纤维化的作用

Objective To investigate whether low-protein diet has protective effect on the progression of renal interstitial fibrosis in rats with cyclosporine A (CsA)-induced nephropathy. Methods Eighteen male Sprague-Dawley rats were randomly divided into three groups (6 rats in each group). The rats in control group (C group) received common diet; in model group (M group) low-salt diet; in intervention group (Ⅰ group) low-salt and low-protein diet. After diet adaptation period of one week, the rats in C group received subcutaneous injection of olive oil 1 mg/kg daily for 5 weeks, while M group and Ⅰ group subcutaneous injection of CsA (diluted into 25 g/L with olive oil) 1 ml/kg for 5 weeks. All the rats were sacrificed at the end of the 5th week. The food-intake and body weight were measured daily. The creatinine clearance (Ccr) was examined before rats were sacrificed. The semi-quantitative pathological analysis on kidney sections was performed. The mRNA and protein expression of transforming growth factor-β1 (TGF-βI) and type Ⅰ collagen (Col Ⅰ) in kidney tissue was determined with real time PCR and immunohistochemical staining, respectively. Results The food-intake and body weight of rats in M and I groups were significantly lower than those in C group (P<0.05). Compared with C group, the Ccr levels in M and Ⅰ groups were significantly reduced [(0.65±0.15) ml/min, (0.40+0.13) ml/min vs (1.55±0.29) ml/min, P<0.05], the relative fibrosis areas of kidney interstitium in M and I groups were significantly increased (3.60%±0.46%, 3.26%±0.75% vs 0.44%±0.24%, P<0.05), the mRNA and protein expression of TGF-β1 in M and I group was significantly up-regulated (by 2.6 and 3.1 times in mRNA and by 1.5 and 1.6 times in protein, respectively, P<0.05), and the mRNA and protein expression of Col Ⅰ in M and I groups was also significantly up-regulated (by 3.0 and 3.5 times in mRNA and by 2.3 and 2.1 times in protein, respectively, P<0.05). There were no significant differences between M and I groups in every parameters above-mentioned except the rat body weight and Ccr. Both the body weight and Ccr in Ⅰ group were significantly lower than those in M group (P<0.05). Compared with C group, the urine osmotic pressure in M group and in I group were deceased (for M group, P>0.05; for I group, P<0.05). Compared with C group, the serum cholesterol levels in M and I groups were significantly increased (P<0.05), and the serum phosphorus level in I group was significantly decreased (P<0.05). The levels of serum albumin and serum calcium of all three groups had no statistical differences (P>0.05). Conclusion Low-protein diet has no renoprutective effects on the rat model of cyclosporin A nephropathy, on the contrary, may induce body weight loss.     

低蛋白饮食对环孢素A肾病大鼠肾间质纤维化的作用

Objective To investigate whether low-protein diet has protective effect on the progression of renal interstitial fibrosis in rats with cyclosporine A (CsA)-induced nephropathy. Methods Eighteen male Sprague-Dawley rats were randomly divided into three groups (6 rats in each group). The rats in control group (C group) received common diet; in model group (M group) low-salt diet; in intervention group (Ⅰ group) low-salt and low-protein diet. After diet adaptation period of one week, the rats in C group received subcutaneous injection of olive oil 1 mg/kg daily for 5 weeks, while M group and Ⅰ group subcutaneous injection of CsA (diluted into 25 g/L with olive oil) 1 ml/kg for 5 weeks. All the rats were sacrificed at the end of the 5th week. The food-intake and body weight were measured daily. The creatinine clearance (Ccr) was examined before rats were sacrificed. The semi-quantitative pathological analysis on kidney sections was performed. The mRNA and protein expression of transforming growth factor-β1 (TGF-βI) and type Ⅰ collagen (Col Ⅰ) in kidney tissue was determined with real time PCR and immunohistochemical staining, respectively. Results The food-intake and body weight of rats in M and I groups were significantly lower than those in C group (P<0.05). Compared with C group, the Ccr levels in M and Ⅰ groups were significantly reduced [(0.65±0.15) ml/min, (0.40+0.13) ml/min vs (1.55±0.29) ml/min, P<0.05], the relative fibrosis areas of kidney interstitium in M and I groups were significantly increased (3.60%±0.46%, 3.26%±0.75% vs 0.44%±0.24%, P<0.05), the mRNA and protein expression of TGF-β1 in M and I group was significantly up-regulated (by 2.6 and 3.1 times in mRNA and by 1.5 and 1.6 times in protein, respectively, P<0.05), and the mRNA and protein expression of Col Ⅰ in M and I groups was also significantly up-regulated (by 3.0 and 3.5 times in mRNA and by 2.3 and 2.1 times in protein, respectively, P<0.05). There were no significant differences between M and I groups in every parameters above-mentioned except the rat body weight and Ccr. Both the body weight and Ccr in Ⅰ group were significantly lower than those in M group (P<0.05). Compared with C group, the urine osmotic pressure in M group and in I group were deceased (for M group, P>0.05; for I group, P<0.05). Compared with C group, the serum cholesterol levels in M and I groups were significantly increased (P<0.05), and the serum phosphorus level in I group was significantly decreased (P<0.05). The levels of serum albumin and serum calcium of all three groups had no statistical differences (P>0.05). Conclusion Low-protein diet has no renoprutective effects on the rat model of cyclosporin A nephropathy, on the contrary, may induce body weight loss.     

低蛋白饮食对环孢素A肾病大鼠肾间质纤维化的作用

Objective To investigate whether low-protein diet has protective effect on the progression of renal interstitial fibrosis in rats with cyclosporine A (CsA)-induced nephropathy. Methods Eighteen male Sprague-Dawley rats were randomly divided into three groups (6 rats in each group). The rats in control group (C group) received common diet; in model group (M group) low-salt diet; in intervention group (Ⅰ group) low-salt and low-protein diet. After diet adaptation period of one week, the rats in C group received subcutaneous injection of olive oil 1 mg/kg daily for 5 weeks, while M group and Ⅰ group subcutaneous injection of CsA (diluted into 25 g/L with olive oil) 1 ml/kg for 5 weeks. All the rats were sacrificed at the end of the 5th week. The food-intake and body weight were measured daily. The creatinine clearance (Ccr) was examined before rats were sacrificed. The semi-quantitative pathological analysis on kidney sections was performed. The mRNA and protein expression of transforming growth factor-β1 (TGF-βI) and type Ⅰ collagen (Col Ⅰ) in kidney tissue was determined with real time PCR and immunohistochemical staining, respectively. Results The food-intake and body weight of rats in M and I groups were significantly lower than those in C group (P<0.05). Compared with C group, the Ccr levels in M and Ⅰ groups were significantly reduced [(0.65±0.15) ml/min, (0.40+0.13) ml/min vs (1.55±0.29) ml/min, P<0.05], the relative fibrosis areas of kidney interstitium in M and I groups were significantly increased (3.60%±0.46%, 3.26%±0.75% vs 0.44%±0.24%, P<0.05), the mRNA and protein expression of TGF-β1 in M and I group was significantly up-regulated (by 2.6 and 3.1 times in mRNA and by 1.5 and 1.6 times in protein, respectively, P<0.05), and the mRNA and protein expression of Col Ⅰ in M and I groups was also significantly up-regulated (by 3.0 and 3.5 times in mRNA and by 2.3 and 2.1 times in protein, respectively, P<0.05). There were no significant differences between M and I groups in every parameters above-mentioned except the rat body weight and Ccr. Both the body weight and Ccr in Ⅰ group were significantly lower than those in M group (P<0.05). Compared with C group, the urine osmotic pressure in M group and in I group were deceased (for M group, P>0.05; for I group, P<0.05). Compared with C group, the serum cholesterol levels in M and I groups were significantly increased (P<0.05), and the serum phosphorus level in I group was significantly decreased (P<0.05). The levels of serum albumin and serum calcium of all three groups had no statistical differences (P>0.05). Conclusion Low-protein diet has no renoprutective effects on the rat model of cyclosporin A nephropathy, on the contrary, may induce body weight loss.     

低蛋白饮食对环孢素A肾病大鼠肾间质纤维化的作用

Objective To investigate whether low-protein diet has protective effect on the progression of renal interstitial fibrosis in rats with cyclosporine A (CsA)-induced nephropathy. Methods Eighteen male Sprague-Dawley rats were randomly divided into three groups (6 rats in each group). The rats in control group (C group) received common diet; in model group (M group) low-salt diet; in intervention group (Ⅰ group) low-salt and low-protein diet. After diet adaptation period of one week, the rats in C group received subcutaneous injection of olive oil 1 mg/kg daily for 5 weeks, while M group and Ⅰ group subcutaneous injection of CsA (diluted into 25 g/L with olive oil) 1 ml/kg for 5 weeks. All the rats were sacrificed at the end of the 5th week. The food-intake and body weight were measured daily. The creatinine clearance (Ccr) was examined before rats were sacrificed. The semi-quantitative pathological analysis on kidney sections was performed. The mRNA and protein expression of transforming growth factor-β1 (TGF-βI) and type Ⅰ collagen (Col Ⅰ) in kidney tissue was determined with real time PCR and immunohistochemical staining, respectively. Results The food-intake and body weight of rats in M and I groups were significantly lower than those in C group (P<0.05). Compared with C group, the Ccr levels in M and Ⅰ groups were significantly reduced [(0.65±0.15) ml/min, (0.40+0.13) ml/min vs (1.55±0.29) ml/min, P<0.05], the relative fibrosis areas of kidney interstitium in M and I groups were significantly increased (3.60%±0.46%, 3.26%±0.75% vs 0.44%±0.24%, P<0.05), the mRNA and protein expression of TGF-β1 in M and I group was significantly up-regulated (by 2.6 and 3.1 times in mRNA and by 1.5 and 1.6 times in protein, respectively, P<0.05), and the mRNA and protein expression of Col Ⅰ in M and I groups was also significantly up-regulated (by 3.0 and 3.5 times in mRNA and by 2.3 and 2.1 times in protein, respectively, P<0.05). There were no significant differences between M and I groups in every parameters above-mentioned except the rat body weight and Ccr. Both the body weight and Ccr in Ⅰ group were significantly lower than those in M group (P<0.05). Compared with C group, the urine osmotic pressure in M group and in I group were deceased (for M group, P>0.05; for I group, P<0.05). Compared with C group, the serum cholesterol levels in M and I groups were significantly increased (P<0.05), and the serum phosphorus level in I group was significantly decreased (P<0.05). The levels of serum albumin and serum calcium of all three groups had no statistical differences (P>0.05). Conclusion Low-protein diet has no renoprutective effects on the rat model of cyclosporin A nephropathy, on the contrary, may induce body weight loss.     

A型肉毒毒素局部应用对兔耳增生性瘢痕创面愈合和瘢痕增生的影响

Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more fibroblasts were in G2-M in gToup S and fewer in G0-G1 (P <0.05). Conclusions Local injection of Botox A can inhibite the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen Ⅰ and Ⅲ in hypertrophic scar, as well as the ratio of collagen Ⅰ to Ⅲ. It serves as the basis for the treatment of hypertrophic scar with Botox A.     

A型肉毒毒素局部应用对兔耳增生性瘢痕创面愈合和瘢痕增生的影响

Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more fibroblasts were in G2-M in gToup S and fewer in G0-G1 (P <0.05). Conclusions Local injection of Botox A can inhibite the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen Ⅰ and Ⅲ in hypertrophic scar, as well as the ratio of collagen Ⅰ to Ⅲ. It serves as the basis for the treatment of hypertrophic scar with Botox A.     

大鼠慢性皮肤溃疡创面愈合中转化生长因子-β1、胶原Ⅰ、胶原Ⅲ的蛋白表达

目的 观察大鼠慢性皮肤溃疡创面愈合过程中转化生长因子-β1( TGF-β1)、胶原Ⅰ和胶原Ⅲ的蛋白表达。方法 将24只8周龄雌性Wister大鼠分为单纯创面组(A组)和皮瓣+创面组即缺血模型组(B组),每组各12只;苏木素-伊红(HE)染色法观察创面1、3、7、10d上皮化率、收缩率及中性粒细胞;采用酶联免疫吸附试验(ELISA)方法测定创面1、3、7、10 d TGF-β1、Ⅰ型和Ⅲ型胶原的蛋白表达。结果 A组上皮化率在各个时间段均高于B组,且在第7天差异有统计学意义(P＜0.05)。A组收缩率明显低于B组。A组中性粒细胞第1、3天逐渐增加,第3天增加到最多,随后逐渐减少;B组在1、3、7d出现增加趋势,第7天增加到最多,第10天减少。TGF-31含量A组于术后1、3、7、10d呈曲线上升趋势,B组在术后1、3、7d逐渐减低,10 d较7d略有回升,且在第1天两组差异有统计学意义(P＜0.05)。胶原Ⅰ蛋白的含量两组随着术后时间的延长均呈减少趋势,在第10天两组差异有统计学意义(P＜0.05)。胶原Ⅲ蛋白的含量两组随术后时间的延长也呈减少的趋势,但在第3天A组比B组明显增加,差异有统计学意义(P＜0.05)。结论 在缺血的干预因素作用下TGF-β1、Ⅰ型和Ⅲ型胶原蛋白表达的减少可能延迟了慢性创伤的正常愈合。