ABSTRACT
AIM: To analyze the relationship between rs128912 single nucleotide polymorphism(SNP)in the promoter region of Toll-like receptor 3(TLR3)gene and cataract in Chinese Han population.METHODS: A total of 263 patients with cataract admitted to our hospital from June 2019 to June 2021 were selected as study group, and 150 patients with lens dislocation were included in control group. Western blotting was used to detect the expression of TLR3 protein in the anterior capsular tissues of lens in the two groups, and direct sequencing method was applied to analyze the polymorphism of rs128912 locus in the promoter region of TLR3 gene. The expression of peripheral blood TLR3 mRNA of patients with different genotypes was detected by real-time quantitative polymerase chain reaction(RT-qPCR).RESULTS: The expression level of TLR3 protein in the anterior capsular tissues in the study group was higher than that in the control group(P<0.05). The frequencies of genotypes(AA, AT, TT)at rs128912 locus in the TLR3 gene promoter region in the study group and the control group were in accordance with Hardy-Weinberg genetic equilibrium, and there were differences in the frequencies of genotypes(AA, AT, TT)and frequencies of alleles(A, T)at rs128912 locus in the TLR3 gene promoter region between both groups(P<0.05). The relative expression level of peripheral blood TLR3 mRNA in patients with TT genotype in the study group was higher than that in patients with AA or AT genotypes(P<0.05).CONCLUSION: The expression of TLR3 protein in anterior capsular tissues of lens of patients with cataract is significantly up-regulated, and rs128912 locus polymorphism in the TLR3 gene promoter region is related to the susceptibility of cataract in Chinese Han population, and people with TT genotype are more prone to cataract.
ABSTRACT
<p><b>OBJECTIVE</b>To study the effect of extract of Radix Tetrastigma hemsleyani on the proliferation and apoptosis of human lung carcinoma H1299 cells, and to explore its mechanisms. METHODS H1299 cells were treated with the extract of Radix Tetrastigma hemsleyani in different concentrations at different time points. Its inhibition on H1299 cell proliferation was detected by Cell Counting Kit-8 (CCK-8) assay. The morphology of the H1299 cell was observed by inverted microscope. Changes of apoptosis were observed by Hoechst33258 methods. The apoptosis rate was detected by flow cytometry. Expression changes of apoptosis-related proteins pro-caspase-3, pro-caspase-9, cle-caspase-3, cle-caspase-9, and poly-ADP-ribose polymerase (PARP) were detected by Western blot.</p><p><b>RESULTS</b>Compared with the control group, the inhibition rate of H1299 cells increased after acted by 0.5, 1, 5, and 10 mg/mL extract of Radix Tetrastigma hemsleyani (P < 0.05, P < 0.01). The longer the acting time, the higher the inhibition rate (P < 0.01). Under inverted microscope, typical morphological changes could be seen and the number of H1299 cells was reduced. Under fluorescence microscope, dark stained nucleus and formed apoptotic body could be observed. Results of flow cytometry showed that the apoptosis rate was obviously dose-effect correlated with the concentration of extract of Radix Tetrastigma hemsleyani. Results of Western blot indicated that compared with the control. group, the protein expression of pro-caspase-3, pro-caspase-9, and PARP were down-regulated and that of cle-caspase-3, cle-caspase-9, and cle-PARP were up-regulated by 5 and 10 mg/mL extract of Radix Tetrastigma hemsleyani (P < 0.05).</p><p><b>CONCLUSIONS</b>Extract of Radix Tetrastigma hemsleyani had obvious effect in inhibiting the proliferation and inducing apoptosis of human lung carcinoma H1299 cells, which might be achieved by activating the expression of caspase protein.</p>