Evidence-based Analysis Of Phototherapy For Melasma And Inhibition Of Skin Inflammatory Response By Ganoderma Lucidum Polysaccharides After Laser Operation

Background:Melasma is a common acquired skin disease with increased pigmentation.Its clinical manifestations are dark brown to dark brown patches on the face,often symmetrically distributed with irregular edges.In addition to the face,melasma may also affect the neck,forearm and chest.Generally more in reproductive age women,especially with dark skin photosensitive female majority.As a kind of volume loss disease,chloasma has a great impact on aesthetics.Although there are a variety of treatments and methods,there is still no measure that can completely cure chloasma.In recent years,the study of the application of photoelectric therapy to treat chloasma is gradually increasing.There are studies abroad that the application of fractional CO₂laser to treat chloasma has achieved excellent curative effect,but there are few similar reports in China.The primary cause may be post-inflammatory hyperpigmentation（PIH）after fractional CO₂laser surgery,which has a higher incidence in Asians.The pathogenesis of PIH is mainly related to the local inflammatory reaction of skin.Excessive accumulation of leukotriene,thromboxane,interleukin-1α（IL-1α）,interleukin-6（IL-6）,tumor necrosis factor-α（TNF-α）and other inflammatory mediators can increase the activity of melanocytes and promote melanin production.Therefore,the key link of prevention and treatment of PIH is to inhibit local inflammatory response.Ganoderma lucidum is a rare herbal medicine in China.Ganoderma lucidum polysaccharides（GLP）is one of the main active ingredients of Ganoderma lucidum.Our previous work suggests that GLP can inhibit UV-induced skin pigmentation,and can greatly reduce the skin erythema reaction caused by UV-induced exposure in guinea pigs.Therefore,we propose the scientific question whether GLP can inhibit skin inflammation after fractional CO₂laser.Purpose:1.The effectiveness and satisfaction of electro-optical therapy for melasma were comprehensively evaluated based on the results of network meta-analysis.2.To explore the effect and mechanism of GLP on inflammatory response of guinea pig skin after fractional CO₂laser surgery.3.To explore the effects of GLP on inflammatory responses of keratinocyte Ha Ca T,human fibroblast FB and vascular endothelial cell HUVEC induced by lipopolysaccharide.Method:The research work carried out in this subject is as follows:Section 1:1.Through searching various databases,formulate strict inclusion and exclusion criteria,and screen all randomized controlled trials involving photoelectric therapy in the treatment of melasma.2.Perform data extraction on the included documents in accordance with the established document information registration form.3.Use the method of network meta-analysis to comprehensively and systematically evaluate the effectiveness and satisfaction of photoelectric therapy in treating melasma.Section 2:1.Explore the establishment of guinea pig animal model of inflammation after CO₂fractional laser operation.2.Evaluate the effect of GLP on inflammation after CO₂fractional laser surgery by gross observation,skin detector detection and HE slice staining.3.After GLP treated the skin lesions of guinea pigs after carbon dioxide laser surgery,the skin lesions of the control group and the GLP treated group were sequenced by RNA-seq,and the differentially expressed genes were analyzed.Gene ontology（GO）analyses and Kyoto Encyclopedia of Genes and Genomes（KEGG）pathway enrichment were performed to screen the genes and signal pathways that may be affected by GLP.Section 3:1.Explore the effects of lipopolysaccharide and GLP on the proliferation of skin keratinocytes,human dermal fibroblasts and vascular endothelial cells through the CCK-8 detection method.2.To study the effect of GLP on the up-regulation of inflammation-related factors induced by lipopolysaccharide in three kinds of skin cells by q RT-PCR.3.Investigating the effect of GLP on lipopolysaccharide induced upregulation of inflammation-related protein expression levels in Ha Ca T by Western blot.Result:Section 1:1.Through the search and screening of the Cochrane Library,Pub Med,MEDLINE,Embase,Web of Science and other databases,17randomized controlled trials of photoelectric therapy for melasma were finally included,involving 14 treatment measures.2.Network Meta-analysis showed that CO₂fractional laser treatment for melasma had a high ranking in terms of effectiveness and patient satisfaction in terms of surface under the cumulative ranking curve（SUCRA）values,and cluster analysis showed that compared with other interventions,CO₂fractional laser had better efficacy and higher patient satisfaction.Section 2:1.After CO₂fractional laser surgery,the application of GLP could reduce the inflammatory infiltration in the skin lesion area of guinea pigs,and the difference was statistically significant（P<0.05）.2.GLP down-regulated NF-κB,IL-17,Toll-like receptors,TNF and other signaling pathways in guinea pig skin lesions after CO₂fractional laser surgery.Section 3:1.Low concentrations of lipopolysaccharide and GLP had no significant cytotoxic effects on Ha Ca T,FB,and HUVEC.2.GLP could down-regulate the m RNA levels of inflammatory factors IL-1β,IL-6 and TNF-αin lipopolysaccharide-induced Ha Ca T,FB and HUVEC,and the difference was statistically significant（P<0.05）.3.GLP could down-regulate the protein expression levels of NF-κB P65,NF-κB P-P65,IL-1β,IL-6,and TGF-β1 inflammation-related proteins in lipopolysaccharide-induced Ha Ca T.Conclusion:1.The network Meta-analysis showed that the clinical efficacy of CO₂fractional laser treatment for melasma was better and patient satisfaction was higher,and it is expected to be one of the effective choices of photoelectric therapy for melasma.2.After CO₂fractional laser,the application of GLP can reduce the inflammatory infiltration in the lesion area of guinea pigs,and GLP may exert anti-inflammatory effects through NF-κB,IL-17,Toll-like receptors,TNF and other signaling pathways.3.GLP could inhibit the inflammatory response by down-regulating the expression levels of inflammatory factors in Ha Ca T,FB,and HUVEC induced by lipopolysaccharide through inhibiting NF-κB signaling pathway.