| Objective:To explore the anti-inflammatory mechanism of Liangxue Tongyu prescription in regulating SCFAs,the metabolites of intestinal flora.Methos:Improvement of autologous blood injection method is adopted to establish the cerebral hemorrhage rat model,observing after 1d,3d,5d three time points.From ethology,brain water content to observe the of Liangxue Tongyu prescription on cerebral hemorrhage rats damage index.Using pathomorphology to observe the inflammatory response of brain tissue,Using Immunohistochemical network stations to detect the expression of NF-κB of brain tissues,Using RT-PCR to dectet the expression of IL-1β、TNF-α mRNA of brain tissues to research the effect of Liangxue Tongyu prescription on inflammatory factors.Using Gas chromatography-mass spectrometry to detect fecal SCFAs in rats with cerebral hemorrhage to analyze the effect of Liangxue Tongyu prescription on metabolites of intestinal flora in rats with cerebral hemorrhage.Using RT-PCR to dectet the expression of GPR43 mRNA of brain and intestinal tissues,Using ELISA to dectet the content of HDAC6 of brain and intestinal tissues,to elucidate the anti-inflammatory mechanism of Liangxue Tongyu prescription in regulating SCFAs,the metabolite of intestinal flora in rats with cerebral hemorrhage.Results:1(1)Ethology score:the ethology score in 3,5d Liangxue Tongyu prescription group and 5d probiotic group were significantly lower than that in model group(P<0.05),both Liangxue Tongyu prescription and probiotics could improve cerebral hemorrhage rats nerve function defect.(2)Brain tissue water content:the brain tissue water content in 3,5d Liangxue Tongyu prescription group and 5d probiotic group were significantly lower than that in model group(P<0.01,P<0.05),both Liangxue Tongyu prescriptiona and probiotics could reduce brain edema.2(1)Brain tissue inflammatory response:there were more inflammatory cells in the model group than that in sham group,there were fewer inflammatory cells in Liangxue Tongyu prescription group and probiotic group than that in model group.(2)Brain tissue NF-κB:each time point the number of NF-κB positive cells in Liangxue Tongyu prescription group and probiotic group were lower than that in model group(P<0.05,P<0.01),they could reduce the NF-κB expression,playing the role of anti-inflammatory.(3)Brain tissue TNF-α and IL-1βmRNA:each time point the expression of TNF-α mRNA in Liangxue Tongyu prescription group was lower than that in model group(P<0.05,P<0.01),the expression of TNF-α mRNA in 3,5d probiotic group were lower that in the model group(P<0.05);each time point the expression of IL-1βmRNA in Liangxue Tongyu prescription group and probiotic group were lower than that in model group(P<0.05,P<0.01),they could reduce the TNF-α and IL-1βmRNA expression,playing the role of anti-inflammatory.3(1)The ratio of acetic acid content in feces:groups of rats expressed no statistical difference in 1,3d.the ratio of acetic acid content in 5d model group was higher than that in sham group and Liangxue Tongyu prescription group(P<0.05,P<0.01),the prescription could reduce the ratio of acetic acid content in feces,(2)The ratio of propionic acid content in feces;groups of rats expressed no statistical difference in 1,3d,the ratio of propionic acid content in 5d Liangxue Tongyu prescription group was higher than that in model group(P<0.05),the prescription could improve the ratio of propionic acid content in feces.(3)The ratio of butyric acid content in feces:groups of rats expressed no statistical difference in 1,3d,the ratio of butyric acid content in 5d model group was lower than that in sham group(P<0.05),there was no difference between Liangxue Tongyu prescription group,probiotic group and sham group(P>0.05),it indicated that the ratio of fecal butyric acid content decreased after cerebral hemorrhage,the prescription could restore the ratio of butyric acid content in feces.(4)The ratio of acetic acid,propionic acid and butyric acid in feces of rats in each group(D5):the ratio of acetic acid content in model group was higher than that in sham group and Liangxue Tongyu prescription group(P<0.05,P<0.01),the ratio of propionic acid content in Liangxue Tongyu prescription group was higher than that in model group(P<0.05),the ratio of butyric acid content in model group was lower than that in sham group(P<0.05),the prescription could reduce the ratio of acetic acid content,improve the ratio of propionic acid content and restore the ratio of butyric acid content in feces.4(1)Brain tissue GPR43mRNA:groups of rats expressed no statistical difference in 1d,the expression of GPR43mRNA in 3,5d Liangxue Tongyu prescription group and 5d probiotic group were significantly higher than that in model group(P<0.01),both Liangxue Tongyu prescription and probiotics could improve the brain tissue GPR43mRNAexpression.(2)Intestinal tissue GPR43 mRNA:groups of rats expressed no statistical difference in 1,3d,the expression of GPR43mRNA on 3,5d in Liangxue Tongyu group and probiotics group were significantly higher than those in the model group(P<0.01),both Liangxue Tongyu prescription and probiotics could improve the intestinal tissue GPR43mRNA expression(3)Brain tissue HDAC6:each time point the content of HDAC6 in Liangxue Tongyu prescription group were lower than that in model group(P<0.01),the content of HDAC6 in 3,5d probiotics group were lower than that in model group(P<0.01),both Liangxue Tongyu prescription and probiotics could reduce the brain tissue HDAC6 expression.(4)Intestinal tissue HDAC6:the content of HDAC6 in 3,5d Liangxue Tongyu prescription group were lower than that in model group(P<0.01),each time point the content of HDAC6 in probiotics group were lower than that in model group(P<0.01,P<0.05),both Liangxue Tongyu prescription and probiotics could reduce the intestinal tissue HDAC6 expressionConclusions:Liangxue Tongyu prescription could inhibit the expression of inflammatory factors such as NF-κB、TNF-α and IL-1β,alleviate cerebral hemorrhage inflammatory reaction,thus it reduced bleeding in the brain edema and improved nerve function.Liangxue Tongyu prescription could decrease the ratio of acetic acid content,increase the ratio of propionic acid content and restore the ratio of butyric acid content in feces.Liangxue Tongyu prescription could inhibit the inflammatory response of brain tissue and play an anti-inflammatory role by increasing the expression of GPR43 and decreasing the expression of HDAC6. |
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