Relationship between Lacticaseibacillus casei Zhang and NPC1L1 and related proteins in cholesterol stone formation in mice
Objective To investigate the effect of Lacticaseibacillus casei Zhang(L.casei Zhang)on hepatic SR-BI as well as small intestinal NPC1L1 and ABCG8 gene expression during gallbladder cholesterol stone formation in mice.Methods The methodology employed involved the acquisition of eight-week-old female C57BL/6J mice of a mean weight of(18.70±0.89)g.Following their purchase,the mice were subjected to a one-week acclimatisation period and were then weighed and divided into a basal group.The chowder saline control group(G-NS,n=10)and the lithogenic chowder saline control group(L-NS,n=10)were established using a random number table method.The basal chowder L.casei Zhang intervention group was also established using this method(n=10).The remaining mice were allocated to two further groups:the G-L-casei group(n=10)and the lithogenic feed L.casei Zhang intervention group(L-L-casei group,n=10).The G-NS and G-L-casei groups were provided with basal feed,the L-NS and L-L-casei groups were given lithogenic feed,and the G-L-casei and L-L-casei groups were administered L.casei Zhang bacterial solution via gavage.The control group was given an equal volume of saline via gavage once per day for eight weeks.Following an eight-week feeding period,the mice were euthanised,and blood was collected via the medial canthus vein.The levels of triglyceride(TG),serum total choles-terol(CHO),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),and apolipopro-tein(Apo-AI)were then determined for the four groups of mice.The formation of cholesterol stone in the mouse gallblad-der was observed with the naked eye and under a polarised light microscope.Additionally,mouse liver and small intestine tissues were collected for the purpose of detecting the expression levels of SR-BI,NPC1L1 and ABCG8,respectively.Results The incidence of gallbladder stones was observed to be significantly lower in the G-NS and G-L-casei groups,in which no stones were identified,compared to the L-NS and L-L-casei groups,in which stones were present.Furthermore,the stone formation rate in the L-L-casei group was found to be lower than that in the L-NS group.The serum CHO expres-sion level was found to be significantly lower in mice from the L-L-casei group compared to those from the L-NS group(t=3.873,P＜0.05).The serum triglyceride(TG)levels of the mice in the L-L-casei group were found to be significantly lower than those of the mice in the L-NS group(t=3.481,P＜0.01).The serum LDL-C level was found to be lower in mice from the L-L-casei group than in those from the L-NS group(t=3.815,P＜0.01).Similarly,the serum HDL-C level was higher in mice from the L-L-casei group than in those from the L-NS group.A statistically significant difference was ob-served between the two groups(t=3.235,P＜0.0l).Additionally,mice in the G-L-casei group exhibited higher levels of the parameter in question than those in the G-NS group,with a statistically significant difference(t=2.649,P＜0.05).The ser-um Apo-AI level was observed to be higher in the L-L-casei group than in the L-NS group,with a statistically significant difference(t=3.938,P＜0.05).Quantitative polymerase chain reaction(RT-qPCR)analysis of the small intestinal NPC1L1 expression level:the L-L-casei group exhibited a lower level of expression than the L-NS group,with a statistically signi-ficant difference(t=3.147,P＜0.01).Similarly,the G-NS group demonstrated a lower level of expression than the L-NS group,with a statistically significant difference(t=2.537,P＜0.05).RT-qPCR small intestinal ABCG8 expression level:the L-L-casei group exhibited a higher level of ABCG8 expression than the L-NS group,with a statistically significant differ-ence.RT-qPCR small intestinal ABCG8 expression level:the L-L-casei group exhibited a higher level of expression than the L-NS group,with a statistically significant difference.The difference was statistically significant(t=2.829,P＜0.05)in the-NS group.RT-qPCR liver SR-BI expression level:the L-L-casei group exhibited a higher level of ABCG8 expression in the small intestine than the L-NS group,with a statistically significant difference(t=2.446,P＜0.05).Conclusion L.casei Zhang has been demonstrated to be an effective agent for the treatment of hyperlipidaemia in mice.It has been shown to re-duce serum cholesterol,triglyceride and LDL-C levels,while simultaneously increasing serum HDL-C and Apo-AI levels.Furthermore,it has been observed to reduce the expression of NPC1L1 in the small intestine,while simultaneously increas-ing the expression of ABCG8 and SR-BI in the liver.This has been linked to a reduction in the incidence of gallbladder stones in mice.
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