Materials and Methods

Animal modelling and grouping treatment of female Wistar rats 30 (provided by Huakang Animal Experimental Center), SPF (specific pathogen free condition) grade, body weight 200 ± 20g, rats before the experiment to adapt to the new environment for 1 week. According to body weight, they were randomly divided into normal control group, model group and Chinese medicine group, with 10 rats in each group. The experimental reference is based on the fourth group of modeling in the Cavaletti [1] method.

Methods Modeling, model group and each Chinese medicine group were intraperitoneally injected with oxaliplatin 3 mg/kg once every other day for 16 times; the normal control group was intraperitoneally injected with an equal volume of 5% glucose. From the modeling d1, the Chinese medicine group soaked the limbs and tail with a concentration of 0.2g/ml Chinese medicine at room temperature, 60min/d, and the model group was soaked in the limbs and tails at room temperature deionized water.

Drugs and laboratory equipment

Chinese herbal compound (Wenjing Tongluo San): Epimedium, geranium, Chuanwu, Chuanxiong, safflower, water decoction prepared by the Pharmacy Department of China-Japan Friendship Hospital to a specific concentration (100ml of medicine per 20g of raw medicine). Oxaliplatin (import): 50 mg is dissolved in 5% dextrose solution at a final concentration of 1 mg/ml. Von Frey Filament: Stoelting, USA. Intelligent hot and cold plate instrument: Beijing Zhongshi Di Chuang Technology Co., Ltd.

BME-410C automatic thermal pain stimulator: Institute of Biomedical Engineering, Chinese Academy of Medical Sciences. Electronic balance (AD-VENTUER OHAVS AR0604). Life Signal Acquisition System: Basic Laboratory of Beijing University of Chinese Medicine.

1. Behavioral testing mechanical pain sensitivity test: Rats were placed on a metal mesh, covered with a transparent plexiglass cover, first adapted to the environment for 15 min, after the combing and inquiry activities basically disappeared, with 4 g and 15 g filaments (Von-Frey Hairs, VFH) gently stimulate the hind paw of the rat, 5 times on each side of the foot, 2 times stimulation interval 6s, the rats immediately showed rapid lift-up reaction during the stimulation time or when removing the fiber, recorded as positive The response, but the lift-response reaction caused by physical activity was not recorded as a positive reaction. The test completed the recording of the number of hind limb lifts and calculated the probability of positive reaction (the number of positive reactions was 10×100%) [2]. VFH 4g stimulated the foot-to-foot reaction in the plantar, ie, mechanically stimulated sensory hypersensitivity; VFH 15g stimulated the foot-to-foot reaction in the rat's foot, ie mechanically stimulated hyperesthesia.

Cold temperature sensitivity test: Turn on the cold plate switch, set the temperature to 4 °C, and place the rat on the cold plate when the temperature reaches the set temperature. Cover the transparent plexiglass cover and let it adapt for 1 min. The number of foot lifts or lameness in the 5 min period of the rats was recorded, and the foot-lifting reaction caused by physical activity or changing position was not recorded as a positive reaction. Behavioral indicators were measured at the beginning of the experiment and there was no difference between the groups.

Electrophysiological test

The sciatic nerve conduction velocity (SNCV): 0.3% pentobarbital sodium anesthetized rats, used to bend the hair to the left hind limb of the rat, exposed the skin, cut along the posterior edge of the femoral shaft Skin, muscle, carefully peel the fascial muscles, expose the sciatic nerve, use the glass minute to dissect the sciatic nerve, place the stimulating electrode and recording electrode of the vital signal acquisition system on the free sciatic nerve, and record the nerve with medlab6.0 on the connected computer after stimulation. The conduction signal, the difference between the time recorded by the two recording electrodes, that is, the difference between the latency, the nerve conduction velocity v is the distance l between the two recording electrodes divided by the difference t of the latency (v = l / t).

Histopathological observation

At the end of the experiment, 6 rats of sciatic nerve were selected in each group and fixed in 10% formalin fixative. After the material was taken, it was embedded in paraffin, sliced ​​to a thickness of 6 μm, dewaxed after sectioning, rehydrated, HE stained, xylene-transparent post-gum seal, and under the optical microscope (Olym-pus), the morphological changes of the sciatic nerve and dorsal root ganglion were observed. .

Statistical method

Some data were processed and analyzed by SPSSl9.0 software. One-way ANOVA was applied to measure the variance of the data, the non-parametric test was applied with irregular variance, and the non-parametric test was used for the count data.

This experiment preliminarily proved the effectiveness of Wenjing Tongluo Powder in the treatment of rat model of peripheral neurotoxicity induced by oxaliplatin, but the main component of the drug onset and the target of peripheral neurotoxicity induced by oxaliplatin It is not clear that further verification and research are still needed.

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