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1.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 33-40, 2022.
Article in Chinese | WPRIM | ID: wpr-940285

ABSTRACT

ObjectiveTo study the effects of Wendantang on the expression of miRNA-219, N-methyl-D-aspartate receptor 2B (NR2B), disrupted in schizophrenia 1 (DISC1), and Ca2+/calmodulin-dependent protein kinase Ⅱγ (CaMKⅡγ) in the frontal lobe of rats with schizophrenia. MethodSixty rats were randomly divided into six groups, namely normal group, model group, high-, medium-, and low-dose Wendantang groups, and clozapine group, with 10 rats in each group. Rats in high-, medium-, and low-dose Wendantang groups were intragastric with 40, 20, and 10 g·kg-1 Wendantang, and the ones in clozapine group were intragastric with 0.02 g·kg-1 clozapine, those in normal and model group were intragastric with equal volume of normal saline, once a day. After 21 days of administration, rats in all groups except for the normal group were injected with 0.6 mg·kg-1 dizocilpine maleate (MK-801) into the left abdominal cavity for inducing acute schizophrenia. The stereotypic behavior and ataxia in rats were scored according to SAMS and HOFFMAN criteria. The morphological changes in the prefrontal cortex were observed by hematoxylin-eosin (HE) staining. The protein expression levels of NR2B, DISC1 and CaMKⅡγ in the frontal lobe was detected by Western blot. The mRNA expression levels of miRNA-219 was detected by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). ResultCompared with normal group, the model group exhibited significantly increased stereotypic behavior and ataxia scores (P<0.01), karyopyknosis and karyolysis in most neurons of the prefrontal cortex, and down-regulated NR2B, DISC1, and CaMKⅡγ protein expression (P<0.01) and miRNA-219, NR2B, DISC1, and CaMKⅡγ mRNA expression (P<0.01). Compared with model group, Wendantang high-, medium-, and low-doses group lowered the scores of stereotypic behavior and ataxia at 50, 60 mmin(P<0.05,P<0.01). In high- and medium-dose Wendantang groups, the neurons in the prefrontal cortex were densely arranged. The karyopyknosis and karyolysis were alleviated to varying degrees. The NR2B protein expression in the frontal lobe was up-regulated (P<0.01). In the medium- and low-dose Wendantang groups, the DISC1 protein expression in the frontal lobe was up-regulated (P<0.05,P<0.01). Wendantang at each dose significantly increased the CaMKⅡγ protein expression (P<0.05) and miRNA-219, NR2B, DISC1, and CaMKⅡγ mRNA expression in the frontal lobe (P<0.05,P<0.01). ConclusionWendantang improves the scores of stereotypical behavior and ataxia, relieves the karyopyknosis and karyolysis of neurons in the prefrontal cortex, and increases the expression levels of miRNA-219, NR2B, DISC1, and CaMKⅡγ of rats with schizophrenia, so as to alleviate the schizophrenic-like symptoms and schizophrenia.

2.
Chinese Journal of Reparative and Reconstructive Surgery ; (12): 641-646, 2016.
Article in Chinese | WPRIM | ID: wpr-856945

ABSTRACT

OBJECTIVE: To investigate the effect of Smad4 on the fibrosis of tendon derived fibroblasts (TDFs) induced by transforming growth factor β1(TGF-β1) by targeted regulation of miRNA219-5P (miR219-5P). METHODS: The tendons donated by the volunteers were harvested to isolate and culture TDFs. The 3rd generation cells were used for experiment. Chemically synthesized miR219-5P mimics, miR219-5P inhibitor, and negative control sequences were transfected into TDFs. The gene expression of miR219-5P in TDFs was detected by real-time PCR, and the protein expression of Smad4 in TDFs was detected by Western blot at 48 hours after transfection. The combining sites of miR219-5P and Smad4 in 3'UTR district were predicted by informatics software. Wild type and mutant type reporter gene expression vectors were constructed and then targeted verification was carried out by the luciferase reporter gene test. Transfected TDFs were then induced by TGF-β1. The proliferation activity of the cells were measured by the cell counting kit 8 after culturing for 24, 48, and 72 hours. The expressions of fibrosis related proteins in TDFs were detected by Western blot at 72 hours. RESULTS: After TDFs were transfected by miR219-5P mimics, miR219-5P expression was significantly up-regulated, but the expressions of Smad4 was decreased subsequently (P0.05). Cell proliferation and the fibrosis related proteins were increased in TGF-β1 induced TDFs, however, decreased in TGF-β1 induced TDFs after transfected by miR219-5P inhibitor (P<0.01). CONCLUSIONS: miR219-5P can significantly inhibit fibrosis of TDFs induced by TGF-β1 by down-regulating Smad4 expression.

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