| Objective:Zhige oral solution is a well-known Chinese medicine Professor Sun Tongjiao’s self-made prescription for relieving alcoholism based on her years of clinical experience.So as to further explore the mechanism of the prescription’s effect on relieving alcoholism and protecting liver,this experiment established a rat model of alcoholic liver injury,to explore the specific mechanism of Zhige oral solution in improving the inflammatory response of alcoholic liver injury in rats from the perspective of purine receptor P2X7 / Nucleotide binding oligomerization domain like receptor protein 3(P2X7R/NLRP3)signaling pathway.Methods: After one week of adaptive feeding,sixty SPF grade male SD rats were randomly divided into the following five groups(n=12): normal group,alcoholic liver injury model group(model group),high-dose Zhige oral solution group(high-dose group),medium dose Zhige oral solution group(medium dose group)and low-dose Zhige oral solution group(low-dose group).The normal group was given distilled water 10 ml / kg per day,the other 4 groups was given52% Vol Luzhou Lao Jiao white spirit,and the volume was 10 m L/kg.Every six hours the high,middle and low dose groups after the gavage of the liquor,and then the rats were given Zhige oral solution respectively according to the dosage of 10,5 and 2.5 m L/kg per day.After 12 weeks of continuous intervention,the blood of abdominal aorta was taken from anesthetized rats,and then the rats were killed,and the liver was taken out.The expression level of alanine aminotransfErase(ALT),aspartate aminotransferase(AST)and γ-Glutamyltransferase(GGT)in serum were detected by biochemical method.Hematoxylin eosin(HE)staining was used to analyze the pathological changes of rat liver;The changes of P2X7R/NLRP3 pathway related proteins were analyzed by Western blot;The expression of NLRP3 in liver was analyzed by immunohistochemistry;Enzyme linked immunosorbent assay(ELISA)was used to detect the content of interleukin-6(IL-6),interleukin-18(IL-18)and Tumor necrosis factor-α(TNF-α),Nuclear factor-κ B(NF-κ B)in serum of different groups of rats.Results: 1.Comparison of serum markers of liver injury in each group:Compared with the normal group,ALT,AST and GGT were significantly increased in the model group(P < 0.01);Compared with the model group,the above indexes in each treatment group decreased in different degrees,and showed a dose-response relationship.2.Comparison of liver HE staining in each group: Compared with the normal group,HE staining in the model group showed disordered arrangement of liver tissue structure,fuzzy boundary,formation of lipid droplets of different sizes in liver cells,accompanied by infiltration of inflammatory cells and necrosis of some liver cells;Compared with the model group,the pathological damage of liver tissue in each treatment group had different content of recovery.Among them,the recovery of high-dose group was the most obvious,which was close to the normal group,followed by the middle dose group,while the difference between the low-dose group and the model group was not obvious,and fat vacuoles of different sizes could also be observed.3.Relative expression of key proteins in P2X7 R / NLRP3 signaling pathway:Compared with the normal group,the protein contents of P2X7 R,NLRP3,Caspase-1 and ASC in the model group were significantly increased,and the difference was statistically significant(P < 0.01);Compared with the model group,the protein expression levels of P2X7 R,NLRP3,Caspase-1 and ASC in Zhige oral solution groups were decreased in various degrees,and the above indexes in high-dose group were decreased most obvious(P < 0.05 or P < 0.01).Staining protein with the method of Immunohistochemical NLRP3 in the liver showed the same result,compared with the normal group,NLRP3 in the model group was significantly increased(P < 0.01),but after the intervention of Zhige oral solution,it had different degrees of recovery.4.Comparison of serum inflammatory injury related factors in different groups of rats: Compared with the normal group,the content of serum IL-6,IL-18 and TNF-α,NF-κB in the model group and low-dose group were significantly higher;Compared with the model group,the above indexes of Zhige oral solution in high-dose and medium dose groups were decreased in various degrees,and the decrease in high-dose group was the most obvious(P < 0.01),while there was no significant difference between low-dose group and model group(P > 0.05).Conclusion: 1.Alcoholic liver injury model can be established successfully by chronic alcohol gavage;2.Zhige oral solution can significantly improve the inflammatory response induced by alcoholic liver injury in rats,and its mechanism may reduce the production of inflammatory injury related factors in rats,through regulating the expression of P2X7 R / NLRP3 signaling pathway related proteins,so as to play a role in improving alcoholic liver injury. |
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