To explore the regulatory effect of Buyang Huanwu Decoction(补阳还五汤)on phenotypic polarization of microglia in cervical spinal cord injury.
Ninety-six female Sprague-Dawley(SD)rats were randomly divided into sham operation,model,low-dose Buyang Huanwu Decoction and high-dose Buyang Huanwu Decoction groups to establish an acute cervical spinal cord injury model.Motor function recovery of rats was assessed using the Forelimb Locomotor Scale(FLS)score and forelimb grip strength experiment Basso-Beattie-Bresnahan(BBB)score before surgery and at 1,3,7,14,21 and 28 d postoperatively.Animals were sacrificed at 7,28 d postoperatively.Hematoxylin and Eosin(HE)staining was used to evaluate the gross morphology of the injury site.Nissl staining was used to assess neuronal survival,and immunofluorescence staining was used to observe the expression of ionized calcium binding adapter molecule 1(Iba1),inducible nitric oxide synthase(iNOS),arginase 1(Arg1)at 7 days post-injury,and microtubule-associated protein 2(MAP2)at 28 days post-injury.Western blot was used to evaluate the expressions of Iba1,CD86,Arg1 and CD206 at 7 days post-injury,and Enzyme-Linked Immunosorbent Assay(ELISA)was used to measure the levels of tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β).
From day 3 to day 28 postoperatively,the motor function in the Buyang Huanwu Decoction groups began to recover significantly compared to the model group(P<0.05).HE staining showed that compared with that in the model group,the spinal cord tissue structure in the Buyang Huanwu Decoction group was denser,with reduced edema,fewer vacuoles and reduced interstitial spaces,and decreased infiltration of immune/inflammatory cells.Nissl staining showed that the number of neurons in the gray matter was increased in the Buyang Huanwu Decoction groups compared with that in the model group,with some surviving neurons visible.The immunofluorescence staining showed that the fluorescence intensity of MAP2 was significantly increased,the fluorescence intensity of M1-type iNOSwas decreased,and the fluorescence intensity of M2-type Arg1 was increased in the Buyang Huanwu Decoction groups compared with those in the model group(P<0.05).Western blot showed that the expression levels of iNOSand CD86 were significantly decreased,while the expression levels of Arg1 and CD206 were significantly increased in the Buyang Huanwu Decoction groups compared with those in the model group(P<0.05).ELISA showed that the levels of TNF-α,IL-6 and IL-1βwere significantly lower in the Buyang Huanwu Decoction groups than those in the model group(P<0.05).
Buyang Huanwu Decoction can effectively promote the phenotypic transition of microglia from M1 to M2 in rats with cervical spinal cord injury,alleviate neuroinflammatory responses,inhibit secondary injury,and thereby improve motor dysfunction after cervical spinal cord injury.
| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |