RESUMEN
ObjectiveTo explore the possible mechanism of reducing cholecystitis and preventing cholelithiasis by Dahuang Lingxian Formula（大黄灵仙方， DLF）. MethodsFifty SD rats were randomly divided into blank group， model group， DLF group， DLF + blank inhibitor group， and DLF + inhibitor group， with 10 rats in each group. The rat model of cholecystitis was established by lipopolysaccharide （LPS） induction in all the groups except for the blank group. Rats in DLF group， DLF + blank inhibitor group and DLF + inhibitor group received intragastric administration of 320 mg/ （kg·d） of DLF 3 days before the preparation of cholecystitis model， while those in blank group and model group were given 2 ml/100 g of distilled water by gastric， twice a day， for 6 consecutive days. Hematoxylin-eosin （HE） staining was used to observe the histopathologic changes of bile duct tissues in each group. The expression of nuclear factor κB （NF-κB） protein in bile duct tissues was detected by immunohistochemistry. The expression levels of interleukin1β （IL-1β） and tumor necrosis factor α （TNF-α） in serum of rats were detected by enzym-linked immunosorbent assay （ELISA）. The mRNA expression levels of miRNA-30b， transforming growth factor β1 （TGF-β1）， nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3） and bone morphogenetic protein 2 （BMP2） in bile duct tissues were detected by real-time PCR， and the protein expression levels of TGF-β1， NLRP3 and BMP2 were detected by Western blot. ResultsCompared to those in the blank group， the structure of the bile duct in the model group was abnormal， and a large number of lymphocytes， plasma cell infiltration and bile canaliculi dilation were seen in the portal area； the positive expression of NF-κB protein increased； there was nuclear infiltration； the expressions of serum inflammatory factors IL-1β and TNF-α， as well as the mRNA and protein expressions of TGF-β1， NLRP3 and BMP2 in bile duct tissue significantly increased， while the expression level of miRNA-30b significantly decreased （P＜0.01）. Compared to those in the model group， the pathological morphology of the bile ducts in the DLF group and DLF + blank inhibitor group was improved， and the infiltration of inflammatory cells was reduced， with decreased positive expression of NF-κB and nuclear infiltration； expression levels of serum inflammatory factors IL-1β and TNF-α， and the mRNA and protein expressions of TGF-β1， NLRP3 and BMP2 in bile duct tissue decreased， while the expression level of miRNA-30b significantly increased （P＜0.05 or P＜0.01）. Compared to the model group， those indicators in the DLF + inhibitor group was not significantly improved （P＞0.05）. There was no significant difference in the indicators between the DLF group and the DLF + blank inhibitor group （P＞0.05）. ConclusionDLF may play a role in delaying the progression of cholelithiasis by regulating the expression of miRNA-30b and inflammation-fibrosis related factors.
RESUMEN
Objective To investigate the expression levels of scf and c-kit under the regulation of Dahuang Lingxian prescription and the possible mechanism of its effect on gallbladder dynamics, and to provide a theoretical basis for Dahuang Lingxian prescription in preventing the development and recurrence of cholesterol gallstone. Methods A total of 45 specific pathogen-free healthy male guinea pigs were randomly divided into normal group, model group, and traditional Chinese medicine (TCM) group. The guinea pigs in the normal group were fed with normal diet, and those in the model group and the TCM group were fed with high-fat lithogenic diet. After 8 weeks of feeding, 5 guinea pigs were randomly selected from each group, and successful modeling was determined if gallstone was observed with the naked eye in more than 4 guinea pigs. After successful modeling, the guinea pigs in the TCM group were given Dahuang Lingxian prescription by gavage, and those in the model group were given an equal volume of normal saline by gavage. After 8 consecutive weeks of administration by gavage, gallbladder tissue samples were collected, and HE staining was used to observe the pathological changes of gallbladder tissue; Western blot was used to measure the expression level of tumor necrosis factor-α (TNF-α) in gallbladder tissue; immunohistochemistry was used to measure the protein expression levels of scf and c-kit in gallbladder smooth muscle tissue. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the least significant difference multiple comparison method was used for further comparison between two groups. Results HE staining showed marked inflammation of gallbladder tissue in the model group, and compared with the model group, the TCM group had a significantly lower degree of inflammation. Western blot showed that the model group had the highest expression level of TNF-α in gallbladder tissue, followed by the TCM group and the normal group ( P < 0.05); immunohistochemistry showed that compared with the model group, the normal group and the TCM group had significantly higher protein expression levels of scf and c-kit in gallbladder smooth muscle tissue ( P < 0.05). Conclusion Dahuang Lingxian prescription can enhance the dynamic function of the gallbladder, possibly by upregulating the scf/c-kit signaling pathway in interstitial cells of Cajal in gallbladder.