Pharmacodynamic Studies Of Zhuanggu Shenjin Capsule On Lumbar Disc Herniation

Objective:To observe the effects of the two different processed Zhuanggu Shenjin capsule on inflammation, analgesic and lumbar disc herniation（LDH） by using a variety of animal models, in order to investigate its protective effect on the pathological alterations of human LDH disease and to explore its mechanism by using LDH rats model, to evaluate whether there is any improvements of the new process drug compared with the old, and to provide more reliable experimental foundations for the clinical application of Zhuanggu Shenjin capsule.Methods:1. Study of anti-inflammatory effects1.1 Carrageenin-induced rats paw swelling experiment78 Wistar rats, male, 150 180 g, according to body weight were randomly assigned for six groups, control group, Zhuanggu Shenjin capsule new process（ZSC-N） 0.3, 0.6, 1.2 g / kg group, Zhuanggu Shenjin capsule old process（ZSC-O）0.6 g / kg group and positive drug Wantong Jingu tablets（WJT） 0.3 g / kg group.Rats were orally administrated with drugs for 7 consecutive days. On the 7th day,after onset of right hind paw swelling by 1% carrageenin suspension, swelling degrees were measured before injection and every an hour from 1h to 6h after injection.1.2 The permeability of the blood capillary in abdominal cavity experiment72 ICR mouse, female, 18 22 g, according to body weight were randomly assigned for six groups, control group, ZSC-N 0.4, 0.8, 1.6 g / kg group, ZSC-O 0.8g / kg group and positive drug WJT 0.4 g / kg group. Mice were orally administrated with drugs for 7 consecutive days. On the 7th day, Evan’s blue was intravenously injected, and acetic acid solution was injected intraperitoneally at the same time.1.3 Cotton pellet-induced formation of granuloma experiment78 Wistar rats, male, 150 180 g, according to body weight were randomly assigned for six groups, control group, ZSC-N 0.3, 0.6, 1.2 g / kg group, ZSC-O 0.6g / kg group and positive drug WJT 0.3 g / kg group. After planting cotton balls in inguinal, drugs were given to the rats for 7 consecutive days. On the 8th day,granuloma and cotton balls were obtained for the calculation of granuloma index, as well as the organ index including thymus, adrenal glands and spleens.2. Study of the analgesic activity2.1 Writhing experiment72 ICR mouse, female and male as a half, 18 22 g, according to body weight were randomly assigned for six groups, control group, ZSC-N 0.4, 0.8, 1.6 g / kg group, ZSC-O 0.8 g / kg group and positive drug WJT 0.4 g / kg group. After drugs administration for 7 consecutive days, experimental model acetic acid-induced writhing in mice was established to evaluate its analgesic activity, times of mice writhing and latent period were recorded.2.2 Hot-plate experiment72 ICR mouse, female, 18 22 g, according to body weight were randomly assigned for six groups, control group, ZSC-N 0.4, 0.8, 1.6 g / kg group, ZSC-O 0.8g / kg group and positive drug WJT 0.4 g / kg group. Pain threshold was recorded before drug administration and every an hour from 1h to 6h after drug administration respectively to calculate the improvement rate of pain threshold in mice.3. Therapeutic effects on lumbar disc herniation rats and its mechanismThe rats were anesthetized with intraperitoneal injection of 10% hydrate and then fix and disinfect. Make a 4cm cut with L4 5 spike gap as the center, cut layers of the skin and subcutaneous tissue incision, and separat of lumbar paravertebral muscle until lamina and spinous process of exposure. After the operation, the animals were put into a cage. On the left thigh muscle intramuscular injection of penicillin sodium 80 thousand U, 1 times a day, 3 days of continuous injection.Recommended by Siegal neural function grade six scoring method to determine, all the rabbits were on the second postoperative day were suffering from generalconcept of limb neurological function score, visible operation after the model rats were right lower limbs sagging, left lower limb systolic performance, neurological abnormalities were greater than grade 2 and above can be considered a successful modeling.Administrat after 7 days of the model, 1 times a day, 21 days of administration.Twenty-second days after administration, the rats were killed and the abdominal aorta was taken from the abdominal aorta, and the right L5 nerve root was taken:Whole blood low, middle, viscosity at high shear rate, plasma prostaglandin E₂（PGE₂）, 5-hydroxytryptamine（5-HT） content, the content of nerve root tissue histamine（HIS）, optical microscope observation rat right L5 nerve root tissue changes, transmission electron microscopy（TEM） in rats on the right side of the L5 nerve root group fabric changes.Result:1. Compared with model group, ZSC-N 0.6, 1.2g/kg and ZSC-O 0.6g/kg could significantly inhibit carrageenin-induced paw swelling（ P<0.05 or P<0.01）.Compared with sham group, both ZSC-N and op had no remarkable effects on the permeability of the blood capillary in abdominal cavity of mice. Compared with model group, ZSC-N 0.6, 1.2 g/kg, op 0.6g/kg could significantly inhibit granuloma index in rats（P<0.05 or P<0.01）, while there were no significant changes on the thymus index, spleen index and adrenal gland index.2. Compared with sham group, ZSC-N 0.8, 1.6g/kg and ZSC-O 0.8g/kg had notable effect on alleviating acetic acid-induced pain and extending the delitescence（ P<0.05 or P<0.01）. ZSC-N 0.8, 1.6g/kg, ZSC-O 0.8g/kg, could remarkably improve the pain threshold at different time windows.3. Compared with sham group, the BV reaction of 20/s 、 60/s 、 120/s are significantly increased（P<0.05 or P<0.01）. The content of PGE₂、5-HT and HIS were significantly increased（P<0.05 or P<0.01）.The L5 nerve root of model group were significantly pathological damaged. Compared with model group, both ZSC-N and ZSC-O can decrease BV reaction of 20/s、60/s、120/s and the content of PGE₂、5-HT、HIS（P<0.05 or P<0.01）. And reduced pathological damages of L5 nerve root.Conclusion:1. Zhuanggu Shenjin capsule has good anti-inflammatory effects and analgesic activity.2. Zhuanggu Shenjin capsule has good effect on lumbar disc herniation.3. Compared with Zhuanggu Shenjin capsule old process, Zhuanggu Shenjin capsule new process’ effect has improved but not to a significant degree.