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Chinese Journal of Experimental Traditional Medical Formulae ; (24): 32-41, 2021.
Artigo em Chinês | WPRIM | ID: wpr-906016

RESUMO

Objective:To explore the effects of Huatan Tongluo Decoction (HTTLD) on the morphology and function of brain tissues and intestine in rats with cerebral ischemia/reperfusion based on the gut-brain axis. Method:Sixty SPF male rats were randomly divided into a sham operation group, a model group, high- (28.66 g·kg<sup>-1</sup>), medium- (14.33 g·kg<sup>-1</sup>), and low-dose (7.16 g·kg<sup>-1</sup>) HTTLD groups, and an edaravone (4 g·kg<sup>-1</sup>)+<italic>Clostridium butyricum</italic> (5.0×10<sup>8</sup> cfu·mL<sup>-1</sup>) group. The model was established by focal cerebral ischemia/reperfusion in rats. The drugs were administered by gavage. The brain tissue injury was determined by neurological deficit score and 2,3,5-triphenyl tetrazolium chloride (TTC) staining. The effect of cerebral ischemia/reperfusion on intestinal motility was assessed by the propulsion rate of small intestine. The intestinal mucosal cell damage was evaluated by the pathomorphological examination of the duodenal mucosa. Enzyme-linked immunosorbent assay (ELISA) was used to determine the content of <italic>D</italic>-lactate (<italic>D</italic>-LAC), diamine oxidase (DAO), and bacterial endotoxin (lipopolysaccharide, LPS) in serum. Western blot was used to detect the expression of Occludin, Claudin-5, and zonula occludens 1 (ZO-1) in the duodenum. Result:After cerebral ischemia/reperfusion, rats developed neurological deficit symptoms. The neurological deficit score in the model group was higher than that in the sham operation group (<italic>P<</italic>0.01). Compared with the model group, the high- and medium-dose HTTLD groups could relieve the symptoms of neurological deficits and lower neurological deficit scores (<italic>P<</italic>0.01). The results of TTC staining showed that the model group presented obvious infarcts in brain tissues compared with the sham operation group (<italic>P<</italic>0.01). The cerebral infarction volumes of HTTLD groups were reduced compared with that in the model group (<italic>P<</italic>0.01), especially the high-dose HTTLD group, and the effect was dose-dependent. Furthermore, the propulsion rate of small intestine in the model group was significantly reduced compared with that in the sham operation group (<italic>P<</italic>0.01). Compared with the model group, HTTLD groups could increase propulsion rates of small intestine (<italic>P<</italic>0.01), especially the high-dose HTTLD group, and the effect was dose-dependent. After cerebral ischemia/reperfusion, obvious duodenal mucosal damage could be observed, which was relieved after the administration of HTTLD. Western blot results showed that the protein expression of ZO-1, Occludin, and Claudin-5 in the model group was reduced compared with that in the sham operation group (<italic>P<</italic>0.01). Compared with the model group, the HTTLD groups could up-regulate the expression of ZO-1, Occludin, and Claudin-5 to varying degrees (<italic>P<</italic>0.05, <italic>P<</italic>0.01), especially the high-dose HTTLD group. ELISA showed that the serum <italic>D</italic>-LAC, DAO, and LPS of the model group were elevated compared with those in the sham operation group (<italic>P<</italic>0.01). Compared with the model group, the HTTLD groups showed reduced <italic>D</italic>-LAC and DAO (<italic>P<</italic>0.05, <italic>P<</italic>0.01), and the medium- and high-dose HTTLD groups showed reduced LPS (<italic>P<</italic>0.05, <italic>P<</italic>0.01), especially the high-dose HTTLD group. Conclusion:After cerebral ischemia/reperfusion, the rats showed damaged brain tissues, neurological dysfunction, intestinal mucosal injury, weakened intestinal motility, and destroyed the intestinal mucosal barrier. HTTLD can protect against brain-gut axis injury after cerebral ischemia/reperfusion by reducing the damage on brain tissues and gastrointestinal mucosa, relieving the symptoms of neurological deficits, promoting gastrointestinal motility, improving intestinal barrier function, and reducing the release of intestinal bacterial metabolites or poisons.

2.
Chinese Traditional and Herbal Drugs ; (24): 4389-4397, 2019.
Artigo em Chinês | WPRIM | ID: wpr-850850

RESUMO

Objective: To investigate the effects of borneol combined with astragaloside IV (AST IV) and Panax notoginseng saponins (PNS) on promoting the active components into the brain and anti-brain injury through the regulation of transporter proteins of blood-brain barrier (BBB) in the state of cerebral ischemia-reperfusion. Methods: Focal cerebral ischemia-reperfusion model in rats was established, borneol, AST IV, PNS and the combination were administered by gavage, brain infarction rate was evaluated by 2,3,5-triphenyl tetrazolium chloride (TTC) staining, the expressions of efflux proteins such as p-glycoprotein (P-gp), multidrug resistance protein (MRP)-1, MRP-2, MRP-4, MRP-5 and uptake proteins such as organic cation transporter (OCT)-3, organicanion transporting polypep-tides (OATP)-2 in brain tissues were detected by Western blotting. The expressions of multidrug resistance (MDR) such as mdr1a, mdr1b and mrp-1, mrp-2, mrp-4, mrp-5 mRNA in brain tissues were determined by real-time PCR method. Results: The results of TTC staining showed that brain infarct was found after cerebral ischemia-reperfusion. Each drug could significantly reduce brain infarction volume and decrease infarction rate, and the effect of AST IV + PNS was better than that of AST IV and PNS alone, the effect of borneol + AST IV + PNS was better than that of single drug and AST IV + PNS. The results of major efflux proteins and genes detection showed that the protein expressions of P-gp, MRP-1, MRP 2, MRP-4, and MRP-5 were significantly increased in rats after cerebral ischemia-reperfusion. Borneol could significantly down-regulate the expressions of P-gp, MRP-2, MRP-4 proteins, PNS could significantly down-regulate the levels of MRP-4, MRP-5 proteins; AST IV, AST IV + PNS and borneol + AST IV + PNS could significantly down-regulate P-gp, MRP-2, MRP-4, MRP-5 proteins, and the effects of borneol + AST IV + PNS were significantly better than those of single drug and AST IV + PNS; The effects of AST IV + PNS were significantly better than those of AST IV or PNS alone. The results of gene expressions were similar to those of protein expression. The results of major uptake proteins showed that the expression of OCT-3 protein did not change significantly in the model group and drug groups after cerebral ischemia-reperfusion; However, the expression of OATP-2 protein was significantly decreased in the model group. PNS, AST IV + PNS and borneol + AST IV + PNS could significantly up-regulate the expression of OATP-2 protein; Furthermore, the effect of borneol + AST IV + PNS was significantly greater than that of single drug and AST IV + PNS, and the effect of AST IV + PNS was significantly greater than that of AST IV and PNS alone. Conclusion: After cerebral ischemia-reperfusion, brain tissues were damaged, the expressions of major efflux proteins and genes on BBB were significantly increased, while the expression of uptake protein such as OATP-2 was significantly decreased. Borneol combined with AST IV and PNS can enhance the effect of anti-ischemic brain injury, which may be related to the down-regulations of the expressions of efflux proteins such as P-gp, MRP-2, MRP-4, MRP-5 and corresponding genes in BBB, as well as the up-regulation of the expression of uptake proteins such as OATP-2, thus promoting the absorption and the enrichment of borneol, AST IV and the effective components of PNS in brain tissues, playing a better role in pharmacology.

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