Effect of moxibustion at "oppositely-located points" on neurogenic bladder after spinal cord injury and endoplasmic reticulum stress pathway in rats / 中国针灸

OBJECTIVE@#To observe the effect of moxibustion at oppositely-located points "Mingmen" (GV 4) and "Shenque" (CV 8) on the motor function of the hind limbs and bladder function in rats with neurogenic bladder after suprasacral spinal cord injury (SCI), so as to explore the effect of this therapy on bladder tissue apoptosis mediated by endoplasmic reticulum stress pathway.@*METHODS@#Twenty-eight female Wistar rats were randomly divided into a sham-operation group (8 rats) and a model establishment group (20 rats). Using the modified Allen's method, the spinal cord of T10 segment was injured to establish a neurogenic bladder model in the model establishment group. Sixteen rats were modeled successfully and then divided into a model group (8 rats) and a moxibustion group (8 rats). In the moxibustion group, 2 h after consciousness regaining from modeling anesthesia, moxibustion was exerted at "Shenque" (CV 8) and "Mingmen" (GV 4), 2 cones at each acupoint in one intervention. The intervention was administered once every two days and 5-time intervention was required totally. After intervention, Basso, Beattie and Bresnahan locomotor rating scale (BBB) score for the motor function of the hind limbs, and the urodynamics indexes (maximum bladder capacity, urine leakage pressure and bladder compliance) were compared among groups. HE staining method was adopted to observe the morphological changes of bladder tissue. With Western blot method and real-time PCR assay, the protein and mRNA expressions of the endoplasmic reticulum stress-related genes (glucose- regulated protein 78 [GRP78], activating transcription factor 4 [ATF4] and cysteinyl aspartate specific proteinase-12 [Caspase-12]) were determined.@*RESULTS@#The transitional epithelial cells were arranged irregularly, the bladder wall was getting thinner, and the cellular vacuolar degeneration and neutrophil infiltration were found in the model group. Whereas, compared with the model group, in the moxibustion group, the arrangement of transitional epithelial cells was clear and continuous in layers, the cellular vacuolar degeneration was mild and the infiltration presented in a small amount of neutrophil granulocytes. Compared with the sham-operation group, in the model group, the BBB score was reduced (P<0.01), the maximum bladder capacity and bladder compliance were increased (P<0.01), and the protein expression levels of GRP78, ATF4 and Caspase-12, as well as mRNA expressions were all increased (P<0.01). In comparison with the model group, in the moxibustion group, BBB score was increased (P<0.01), the maximum bladder capacity and bladder compliance were decreased (P<0.01), and the protein and mRNA expression levels of GRP78, ATF4 and Caspase-12 were all decreased (P<0.01).@*CONCLUSION@#Moxibustion at the "oppositely-located points" improves the urination function, alleviate urine retention in neurogenic bladder rats after spinal cord injury. The underlying mechanism may be related to the down-regulation of the expressions of GRP78, ATF4 and Caspase-12 in the endoplasmic reticulum stress pathway of the bladder tissues, and thus to alleviate the apoptosis of bladder tissue.

[Preparation of Cangyi nanoemulsion in situ gel and drug release mechanism of nasal mucosa in vitro].

To prepare Cangyi nanoemulsion in situ gel and study its nasal mucosa release mechanism in vitro. After proper treatment of different drugs in the compound, the prescription of nanoemulsion was determined by pseudo-ternary phase diagram method. With the ratio of mixed emulsifier to oil phase [(S+COS)/O], the ratio of mixed emulsifier(K_m), the ratio of water phase to mixed emulsifier and oil phase[W/(S+COS+O)] as investigation factors and the normalized value(OD) as evaluation index, the prescription of Cangyi nanoemulsion was optimized by central composite design-response surface method. With the ratio of poloxamer 407(P407) and poloxamer 188(P188) as the investigation factors and the gelation temperature as the evaluation index, the in situ gel prescription of Cangyi nanoemulsion was optimized. The improved Franz diffusion cell was used to explore the nasal mucosa drug-release mechanism of Cangyi nanoemulsion in situ gel with oxymatrine, ferulic acid and salvianolic acid B content as indexes. The optimal prescription of Cangyi nanoemulsion in situ gel was as follows: 6.862% castor oil polyoxyl(EL), 4.262% absolute ethanol, 1.392% ethyl oleate, 7% P407 and 6% P188. The average pH was 5.55 and the average gelation temperature was 32.8 ℃. In vitro release studies showed that oxymatrine, ferulic acid and salvianolic acid B were released simultaneously and the drug release behavior was consistent with that in Higuchi model. The preparation process of Cangyi nanoemulsion in situ gel is stable, with suitable pH value, gelation temperature and viscosity. It has a certain slow-release effect, and can meet the needs of local nasal drug use.