靳三针对缺血性脑卒中大鼠模型的实验研究
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摘要
目的
课题的具体研究内容：本课题在针刺对中风的文献总结的基础上选择大鼠脑中风模型,以靳三针为主对其进行针刺治疗,以大鼠存活率,神经功能损害程度,血清乳酸脱氢酶含量测定等指标为检测对象,与假手术组,模型组相比较,为针刺治疗脑中风的作用研究提供一定的实验依据。
方法
将大鼠分为假手术组、针刺组、模型组3组动物。针刺组与模型组均以线栓法阻断一侧MCA,术中维持体温37±5℃。大鼠用10%水合氯醛3ml/kg腹腔麻醉,颈部正中切开,分离左侧颈总动脉、颈外动脉、颈内动脉和颈内动脉颈外分支一翼腭动脉。结扎颈总动脉近心端、颈外动脉起始端和翼腭动脉起始端后,经颈总动脉插入线栓,线栓采用单股尼龙线,预先在尼龙线上作好标记刻度,头端磨钝,并用10%L-赖氨酸浸泡。以颈动脉叉为起始标记,插入深度为1.8±0.5cm,到有轻微阻力感为止,扎紧动脉残端,缝合皮肤。假手术组操作与上述3组操作相同,但不插线。术后动物单笼饲养。
筛选造模成功者,在术后24h进行针刺治疗。本次实验全部穴位均按照华兴邦等制定的《动物针灸穴位图谱》取穴。针刺组以颞三针为主穴,配以手三针、足三针,由同一人扎针,均留针30min,其间每10min行针1次。隔天针刺,共针3次。常规组取穴：上肢取肩骨禺、曲池、外关、合谷,下肢取环跳、阳陵泉、足三里、解溪、昆仑,手法采用针刺患侧,用提插捻转补法各1min。模型组不予以针刺,进行临床体征观察、大鼠存活率及神经症状,脑组织含水量及水肿程度、血清乳酸脱氢酶测定,等观察指标进行观察。
结果
(1)临床体征观察：大鼠体重190-210g,皮毛光滑,口鼻唇红润,精神状态好,好动打架,攻击性强,饮食好,大便干燥。线栓法手术后,大鼠出现毛发枯槁,精神萎靡不振,体重明显下降,活动减少,攻击性减弱,饮食量减少,口鼻色泽黯淡,便溏。手术后出现偏瘫症状：左侧前爪软弱无力、左前爪不能内收,爬行时向左侧转圈,身体向左侧倾斜。(2)大鼠存活率及神经症状：针刺组大鼠存活率在手术后三次治疗时,都明显高于模型组,假手术组无神经功能缺损,模型组与针刺组缺血2h神经功能缺损比较无显著性差异((P>0.05)。模型组于再灌注24h后神经功能缺损较缺血2h加重,针刺组大鼠于再灌注24h神经功能缺损即有所恢复,于各时间观察点与模型组比较有显著性差异(P<0.05)。(3)脑组织含水量及水肿程度：模型组脑组织含水量明显增高与其他组,表明造模后可导致脑含水量增加；针刺量组含水量比模型组低。(4)血清乳酸脱氢酶(LDH)测定：一周后,模型组LDH含量比假手术组显著升高(p<0.5),针刺组LDH含量明显低于同时段模型组(p<0.01)。(5)血清aCL—IgG水平测定结果显示模型组动物血清aCL—IgG水平含量比假手术组显著升高(p<0.05),进一步说明手术造模成功。针刺组aCL-IgG水平含量明显低于模型组(p<0.01)。(6)血清纤维蛋白原测定结果显示模型组动物血清纤维蛋白原含量比假手术组显著升高(p<0.05),进一步说明手术造模成功。针刺组纤维蛋白原含量与模型组相比有降低的趋势,但差异并不显著(p>0.05)。
结论
线栓法大鼠中风模型具有缺血性中风的症状,手术后,模型大鼠出现不同程度的偏瘫症状：左侧前爪软弱无力、左前肢不能内收,爬行时向左侧转圈,甚至身体向左侧倾斜。操作简单,造模亦成功。针刺对局灶性脑缺血大鼠的神经功能具有保护作用,能降低大鼠死亡率,改善神经症状,脑组织含水量及水肿程度降低,血清乳酸脱氢酶亦降低.说明针刺在治疗和改善脑中风方面有很好的效果。
Purpose
The detail of this study:the subject is selected on the basis of literature, rat stroke models were used to measure the efficacy of actupuncture especially the JIN'3needle on the stroke. Rat survival, the extent of neurological function's damage, serum lactate dehydrogenase and others were compared between the conventional group, model group to provide experimental evidence for acupuncture treatment of stroke research.
Methods
The rats were divided into conventional group, acupuncture group and model group. Acupuncture group and model groups were used line leash method to suture the MCA to make stroke modle, during the period of surgery, maintain body temperature at37±5℃.10%chloral hydrate in the dosage of3ml/kg in the way of intraperitoneal to anesthesia the rats, neck midline incised, separated of the left common carotid artery, external carotid artery, internal carotid artery and internal carotid artery external carotid branches pterygopalatine artery. Ligation of the common carotid artery proximal end, the starting end of the external carotid artery and pterygopalatine artery end, the bolt of the common carotid artery was cannulated line, the line tied with a single strand of nylon line, pre-tagged scale in the nylon line head-end mill obtuse, and soaked with10%L-lysine. Carotid fork as the starting mark the insertion depth was1.8±0.5cm, to date there was a slight sense of resistance, truss artery stump skin was sutured. Sham operation group operation and the operation is the same, but do not plug wire. Postoperative animals caged with.
Screening modeling successful in24h after acupuncture treatment. This experiment, all other points in accordance with the Huaxing Bang "animal acupuncture points map" acupoints. Acupuncture group to the temporal three-pin main points with three-pin of hand, foot three needles, acupuncture treatment by the same person, the needle for30min, during which every10min line pin1. Acupuncture every other day, to pin3. The conventional group acupoints:upper extremity of shoulder skull, Qu pool, foreign relations, gu, lower extremity take Huantiao, Yang Ling Quan, Zusanli, Xie River, Kunlun, the way the use of acupuncture affected side, with lift and thrust twisting reinforcing each1min. Model group by acupuncture, clinical signs observed, the survival rate of rats and neurological symptoms, brain water content and the degree of edema, serum lactate dehydrogenase determination, were used in this observation.
Results
clinical signs observed:body weight190-210g, fur smooth, rosy lips nose and mouth, a good mental state, active fighting, aggressive, good diet, dry stools. The suture method after surgery, the rats withered hair, spiritual malaise, weight decreased, reduced activity, offensive weakened, reduced food intake, pale green nose and mouth, loose stools. Post-operative the hemiplegia symptoms:the left front paw is weak, the left front paw is not received within a circular motion to the left, crawling, body tilted to the left.(2) rat survival and neurological symptoms:three treatment acupuncture group survival rate after surgery was significantly higher than the model group, sham operation group without neurological deficit, model group and acupuncture group ischemia2h nerve function defect was no significant difference ((P>0.05). model group at24h after reperfusion neurological deficit compared with ischemia2h aggravated the acupuncture group reperfusion24h neurological deficit that has been restored, at each time observation points compared with model group with significant difference (P<0.05)(3) brain tissue water content and the degree of edema:model group brain water content was significantly higher with the other groups, indicating that the model can lead to an increase in brain water content; Acupuncture amount of group water content lower than the model group.(4) Determination of serum lactate dehydrogenase (LDH):A week later, the LDH content of the model group than the sham-operated group was significantly higher (p<0.5), the LDH content of the acupuncture group lower than the same period the model group (p<0.01).(5) serum ACL-IgG levels were measured results show that the model group level of serum aCL-IgG content was significantly higher than in the sham-operated group (p<0.05), further surgical modeling success. Acupuncture group aCL-IgG level was significantly lower than the model group (p<0.01).(6) Determination of serum fibrinogen results show that the model group animal serum fibrinogen was significantly higher than in the sham-operated group (p<0.05), further surgery successful modeling. Fibrinogen content of the acupuncture group compared with model group tended to decrease, but the difference was not significant (p>0.05).
Conclusion
The method of line leash to make ischemic stroke rat models, stroke symptoms were appeared after surgery, the rat model of different degrees of paralysis symptoms:the left front paw is weak, the left forelimb can not adduct circling to the left, crawling, even the body tilted to the left. This method was Simple operation, and modle easily success. Acupuncture has a protective effect on focal cerebral ischemia neurological, can reduce rat mortality, improve neurological symptoms, reduced brain water content and the degree of edema, serum lactate dehydrogenase is also reduced. Acupuncture can treat and have a good effect to improve stroke aspect.
课题的具体研究内容：本课题在针刺对中风的文献总结的基础上选择大鼠脑中风模型,以靳三针为主对其进行针刺治疗,以大鼠存活率,神经功能损害程度,血清乳酸脱氢酶含量测定等指标为检测对象,与假手术组,模型组相比较,为针刺治疗脑中风的作用研究提供一定的实验依据。
方法
将大鼠分为假手术组、针刺组、模型组3组动物。针刺组与模型组均以线栓法阻断一侧MCA,术中维持体温37±5℃。大鼠用10%水合氯醛3ml/kg腹腔麻醉,颈部正中切开,分离左侧颈总动脉、颈外动脉、颈内动脉和颈内动脉颈外分支一翼腭动脉。结扎颈总动脉近心端、颈外动脉起始端和翼腭动脉起始端后,经颈总动脉插入线栓,线栓采用单股尼龙线,预先在尼龙线上作好标记刻度,头端磨钝,并用10%L-赖氨酸浸泡。以颈动脉叉为起始标记,插入深度为1.8±0.5cm,到有轻微阻力感为止,扎紧动脉残端,缝合皮肤。假手术组操作与上述3组操作相同,但不插线。术后动物单笼饲养。
筛选造模成功者,在术后24h进行针刺治疗。本次实验全部穴位均按照华兴邦等制定的《动物针灸穴位图谱》取穴。针刺组以颞三针为主穴,配以手三针、足三针,由同一人扎针,均留针30min,其间每10min行针1次。隔天针刺,共针3次。常规组取穴：上肢取肩骨禺、曲池、外关、合谷,下肢取环跳、阳陵泉、足三里、解溪、昆仑,手法采用针刺患侧,用提插捻转补法各1min。模型组不予以针刺,进行临床体征观察、大鼠存活率及神经症状,脑组织含水量及水肿程度、血清乳酸脱氢酶测定,等观察指标进行观察。
结果
(1)临床体征观察：大鼠体重190-210g,皮毛光滑,口鼻唇红润,精神状态好,好动打架,攻击性强,饮食好,大便干燥。线栓法手术后,大鼠出现毛发枯槁,精神萎靡不振,体重明显下降,活动减少,攻击性减弱,饮食量减少,口鼻色泽黯淡,便溏。手术后出现偏瘫症状：左侧前爪软弱无力、左前爪不能内收,爬行时向左侧转圈,身体向左侧倾斜。(2)大鼠存活率及神经症状：针刺组大鼠存活率在手术后三次治疗时,都明显高于模型组,假手术组无神经功能缺损,模型组与针刺组缺血2h神经功能缺损比较无显著性差异((P>0.05)。模型组于再灌注24h后神经功能缺损较缺血2h加重,针刺组大鼠于再灌注24h神经功能缺损即有所恢复,于各时间观察点与模型组比较有显著性差异(P<0.05)。(3)脑组织含水量及水肿程度：模型组脑组织含水量明显增高与其他组,表明造模后可导致脑含水量增加；针刺量组含水量比模型组低。(4)血清乳酸脱氢酶(LDH)测定：一周后,模型组LDH含量比假手术组显著升高(p<0.5),针刺组LDH含量明显低于同时段模型组(p<0.01)。(5)血清aCL—IgG水平测定结果显示模型组动物血清aCL—IgG水平含量比假手术组显著升高(p<0.05),进一步说明手术造模成功。针刺组aCL-IgG水平含量明显低于模型组(p<0.01)。(6)血清纤维蛋白原测定结果显示模型组动物血清纤维蛋白原含量比假手术组显著升高(p<0.05),进一步说明手术造模成功。针刺组纤维蛋白原含量与模型组相比有降低的趋势,但差异并不显著(p>0.05)。
结论
线栓法大鼠中风模型具有缺血性中风的症状,手术后,模型大鼠出现不同程度的偏瘫症状：左侧前爪软弱无力、左前肢不能内收,爬行时向左侧转圈,甚至身体向左侧倾斜。操作简单,造模亦成功。针刺对局灶性脑缺血大鼠的神经功能具有保护作用,能降低大鼠死亡率,改善神经症状,脑组织含水量及水肿程度降低,血清乳酸脱氢酶亦降低.说明针刺在治疗和改善脑中风方面有很好的效果。
Purpose
The detail of this study:the subject is selected on the basis of literature, rat stroke models were used to measure the efficacy of actupuncture especially the JIN'3needle on the stroke. Rat survival, the extent of neurological function's damage, serum lactate dehydrogenase and others were compared between the conventional group, model group to provide experimental evidence for acupuncture treatment of stroke research.
Methods
The rats were divided into conventional group, acupuncture group and model group. Acupuncture group and model groups were used line leash method to suture the MCA to make stroke modle, during the period of surgery, maintain body temperature at37±5℃.10%chloral hydrate in the dosage of3ml/kg in the way of intraperitoneal to anesthesia the rats, neck midline incised, separated of the left common carotid artery, external carotid artery, internal carotid artery and internal carotid artery external carotid branches pterygopalatine artery. Ligation of the common carotid artery proximal end, the starting end of the external carotid artery and pterygopalatine artery end, the bolt of the common carotid artery was cannulated line, the line tied with a single strand of nylon line, pre-tagged scale in the nylon line head-end mill obtuse, and soaked with10%L-lysine. Carotid fork as the starting mark the insertion depth was1.8±0.5cm, to date there was a slight sense of resistance, truss artery stump skin was sutured. Sham operation group operation and the operation is the same, but do not plug wire. Postoperative animals caged with.
Screening modeling successful in24h after acupuncture treatment. This experiment, all other points in accordance with the Huaxing Bang "animal acupuncture points map" acupoints. Acupuncture group to the temporal three-pin main points with three-pin of hand, foot three needles, acupuncture treatment by the same person, the needle for30min, during which every10min line pin1. Acupuncture every other day, to pin3. The conventional group acupoints:upper extremity of shoulder skull, Qu pool, foreign relations, gu, lower extremity take Huantiao, Yang Ling Quan, Zusanli, Xie River, Kunlun, the way the use of acupuncture affected side, with lift and thrust twisting reinforcing each1min. Model group by acupuncture, clinical signs observed, the survival rate of rats and neurological symptoms, brain water content and the degree of edema, serum lactate dehydrogenase determination, were used in this observation.
Results
clinical signs observed:body weight190-210g, fur smooth, rosy lips nose and mouth, a good mental state, active fighting, aggressive, good diet, dry stools. The suture method after surgery, the rats withered hair, spiritual malaise, weight decreased, reduced activity, offensive weakened, reduced food intake, pale green nose and mouth, loose stools. Post-operative the hemiplegia symptoms:the left front paw is weak, the left front paw is not received within a circular motion to the left, crawling, body tilted to the left.(2) rat survival and neurological symptoms:three treatment acupuncture group survival rate after surgery was significantly higher than the model group, sham operation group without neurological deficit, model group and acupuncture group ischemia2h nerve function defect was no significant difference ((P>0.05). model group at24h after reperfusion neurological deficit compared with ischemia2h aggravated the acupuncture group reperfusion24h neurological deficit that has been restored, at each time observation points compared with model group with significant difference (P<0.05)(3) brain tissue water content and the degree of edema:model group brain water content was significantly higher with the other groups, indicating that the model can lead to an increase in brain water content; Acupuncture amount of group water content lower than the model group.(4) Determination of serum lactate dehydrogenase (LDH):A week later, the LDH content of the model group than the sham-operated group was significantly higher (p<0.5), the LDH content of the acupuncture group lower than the same period the model group (p<0.01).(5) serum ACL-IgG levels were measured results show that the model group level of serum aCL-IgG content was significantly higher than in the sham-operated group (p<0.05), further surgical modeling success. Acupuncture group aCL-IgG level was significantly lower than the model group (p<0.01).(6) Determination of serum fibrinogen results show that the model group animal serum fibrinogen was significantly higher than in the sham-operated group (p<0.05), further surgery successful modeling. Fibrinogen content of the acupuncture group compared with model group tended to decrease, but the difference was not significant (p>0.05).
Conclusion
The method of line leash to make ischemic stroke rat models, stroke symptoms were appeared after surgery, the rat model of different degrees of paralysis symptoms:the left front paw is weak, the left forelimb can not adduct circling to the left, crawling, even the body tilted to the left. This method was Simple operation, and modle easily success. Acupuncture has a protective effect on focal cerebral ischemia neurological, can reduce rat mortality, improve neurological symptoms, reduced brain water content and the degree of edema, serum lactate dehydrogenase is also reduced. Acupuncture can treat and have a good effect to improve stroke aspect.
引文
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