接骨胶囊促进骨修复及抗炎镇痛作用的动物实验

An animal experiment on effect of bone-joining capsule in counteracting inflammation ,easing pain and promoting bone repair

目的:探讨接骨胶囊促进家兔骨折愈合及对小鼠的抗炎及镇痛作用。方法:实验于2003-01/12在河南中医学院中心实验室完成。①选择日本大耳家兔56只,分为接骨胶囊高、低剂量组、药物对照组和空白对照组,每组14只,造成双侧桡骨骨缺损后分别给予接骨胶囊0.8,0.4g/kg、麝香接骨丹0.5g/kg和生理盐水10mL/kg。于给药20,40d后,取双侧桡骨样本,应用X射线片及组织学观察评价骨折愈合程度。②选择昆明种小白鼠120只,用于3个实验(毛细血管通透性、耳郭肿胀实验及镇痛实验),每个实验40只。各实验中小鼠分别分为接骨胶囊高、低剂量组、药物对照组和空白对照组,每组10只,分别给予接骨胶囊0.8,0.4g/kg、麝香接骨丹0.5g/kg和生理盐水1mL/只。检测小白鼠腹腔注射醋酸后毛细血管通透性通过测定伊文思蓝吸光度。小白鼠右耳涂抹二甲苯致耳郭肿胀,3h后与左耳对比,以左右两耳片质量差值作为耳肿胀度。小白鼠腹腔注射醋酸后15min内观察疼痛所致扭体次数。结果:家兔56只及小白鼠120只全部进入结果分析,无脱落。①X射线改变:术后各组家兔均较空白对照组改变明显(H=58～92,P<0.05～0.01)。用药40d,各药物组骨折端已由骨痂所充填,进入骨痂改建期,空白对照组的骨折愈合进程明显慢于各药物组。②毛细血管通透性改变:应用伊文思蓝吸光度表示,空白对照组小鼠高于接骨胶囊高剂量组和药物对照组[0.46±0.09,0.28±0.07,0.37±0.10,(t=4.84,2.14,P<0.01,0.05)]。③耳廓肿胀度:接骨胶囊高、低剂量组和药物对照组小鼠均低于空白对照组[(8.51±1.82),(10.42±1.57),(9.72±1.68),(15.89±2.71)mg,(t=7.15,5.52,6.12,P<0.01)]。④扭体次数:空白对照组小鼠高于接骨胶囊高剂量组和药物对照组[(35.6±6.6),(17.1±5.2),(28.7±7.1)次,(t=6.92,2.25,P<0.01,0.05)]。接骨胶囊高、低剂量组和药物对照组的镇痛率分别为51.9%,17.4%和19.4%。结论:接骨胶囊能明显促进家兔骨折愈合;显著抑制醋酸所致的小鼠毛细血管通透性增加及二甲苯所致的小白鼠耳郭肿胀,证明其有抗炎作用;接骨胶囊又可以明显抑制腹腔注射醋酸引起的小白鼠扭体反应,具有镇痛作用。

AIM： To investigate the effect of bone-joining eapsule in promoting fracture healing, counteracting inflammation and easing pain. METHODS： The experiment was conducted in the central laboratory of Henan Hospital of Traditional Chinese Medicine from January to December 2003. ①Fifty-six Japanese flap-eared rabbits were randomly and equally divided into high-dose, low-dose bone-joining capsule, drug control and blank control groups. The rabbits in their respective groups were treated with 0.8, 0.4 g/kg bone-joining capsule, 0.5 g/kg bone-joining musk pill and 10 mL/kg normal saline after they were induced into the models of bilateral radial defect. The bilateral radial bones were sampled to assess the degree of fracture healing by X-ray and histological observation at 20 and 40 days after administration. ②Totally 120 Kunming white mice were selected and used in three experiments on capillary permeability, auricular swelling and analgesia, and 30 mice in each experiment. Each 30 white mice were equally divided into high-dose, low-dose bone-joining capsule, drug control and blank control groups, in which the mice were treated with 0.8, 0.4 g/kg bone-joining capsule, 0.5 g/kg bone-joining musk pill and 1 mL/kg normal saline respectively. The capillary permeability was measured by detecting Evans Blue absorbance after acetic acid was injected intraperitoneally. Dimethylbenzene was daubed onto the right ears of the white mice to induce auricular swelling. Then the swelling right ears were compared with the left ones 3 hours later, and the mass difference of the right ears minus left ears was used to evaluate the degree of auricular swelling. The times of body writhe were observed within 15 minutes after acetic acid was injected intraperitoneally. RESULTS： All the 56 rabbits and 120 white mice were included in the result analysis without loss. ③The results of X-ray observation altered more significantly in rabbits of the drug treatment groups than in those of the blank control group （H=58 to 92,P 〈 0.05 to 0.01）. After 40-day drug treatment, the ends of fractured bone in the drug treatment groups were filled with callus, stepping into the phase of reconstruction by callus, however the speed of fracture healing in the control group was remarkably slower than that in the drug treatment groups. ②Changes of capillary permeahility： The Evans Blue absorbance was significantly higher in the mice of the blank control group than in the high-dose bone-joining capsule group and drug control group[0.46±0.09 vs 0.28±0.07, 0.37±0.10, t=4.84, 2.14, P 〈 0.01, 0.051.③The degree of auricular swelling： The mass difference of the right ears minus left ears was significantly less in the low-dose, high-dose bone-joining capsule and drug control groups than in the blank control group[（8.51±1.82）, （10.42±1.57）, （9.72±1.68） mg vs （15.89±2.71） mg, t=7.15, 5.52, 6.12, P 〈 0.01]. ④The times of body writhe in the blank control group were significantly more than those in the high-dose bone- joining capsule and drug control groups[（35.6±6.6）, （17.1±5.2） times vs （28.7±7.1） times, t=6.92, 2.25, P 〈 0.01, 0.05]. The analgesia rate was 51.9%, 17.4% and 19.4% in the high-dose, low-dose bone-joining capsule and drug control groups respectively. CONCLUSION： Bone-joining capsule can obviously accelerate the fracture healing in rabbits, and inhibit the aggravation of acetic acid-induced capillary permeability and dimethylbenzene-induced auricular swelling in mice, indicating its anti-inflammation effect; Bone-joining capsule can obviously decrease the times of mouse body writhe induced by intraperitoneal injection of acetic acid, so it has analgesic effect.