The Influence Of Moxibustion With Different Temperature On Local Skin Langerhans Cell And Its Correlation With TRPV1

Objective:To observe the influence of moxibustion with different temperature on local skin tissue structure and the distribution,morphology and quantity of Langerhans Cell?LC?in acupoint,and to explore the role of TRPV1 during moxibustion stimulating LC in local skin.By above studying,we hope to investigate the mechanism of moxibustion initiation effect in term of skin immune system.Methods:Wide type C57BL/6J mice?WT?and TRPV1 gene knockout(TRPV1^-/-)mice aged3 mouthes were divided into WT+control group,WT+38?moxi group,WT+46?moxi group,TRPV1^-/-+control group,TRPV1^-/-+38?moxi group,TRPV1^-/-46?+moxi group,with 24 mices in each group.Moxibustion stimulated the“Zusanli?ST36?”on the left leg of mice lasting 30 min and the local skin tissue of 12mices in each group were respectively taken at two time point?30 min and 72 h?after moxibustion.In local skin of C57BL/6J wild type mice and TRPV1^-/-gene knockout mice,the effects of moxibustion on the skin structure were detected by HE staining and on the morphology,quantity and distribution of Langerhans cells were detected by immunohistochemistry and immunofluorescence assay?the expression of CD1a and S100 protein?.Results:1.The effects of moxibustion with different temperature on the skin structure and the LC of local skin in wild type mice:At 30min after moxibustion,compared with the WT+control group,WT+38?moxi group's epidermis was easily peeled off,a small amount of inflammatory cell infiltration,small CD1a⁺and S100⁺LC and its main distribution in epidermis and dermis,no obvious change in LC'S quantity?P>0.05?;WT+46?moxi group's epidermis was extremely easily peeled off,a abundant of inflammatory cell infiltrationand,hair follicle and sebaceous gland with hyperplasia and destruction,smaller CD1a⁺and S100⁺LC and its main distribution in hair follicle and sebaceous gland,obvious increase in LC'S quantity?P<0.05?.The quantitative change of CD1a⁺and S100⁺LC was unobvious between at two?30min,72h?time point after moxibustion in WT+38?moxi group?P>0.05?.Compared with the WT+46?moxi group of 30min after moxibustion,the number of CD1a⁺and S100⁺LC were significantly decreaseed in the one of 72h after moxibustion?P<0.05?.2.The effects of moxibustion with different temperature on the skin structure and the LC of local skin in TRPV1 knockout mice:At 30min after moxibustion,compared with the TRPV1^-/-+control group,TRPV1^-/-+38?moxi group's epidermis was easily peeled off,a small amount of inflammatory cell infiltration,small CD1a⁺and S100⁺LC and its main distribution in the epidermis and dermis,no obvious change in LC'S quantity?P>0.05?.TRPV1^-/-+46?moxi group's epidermis was extremely easily peeled off,a abundant of inflammatory cell infiltration,hair follicle and sebaceous gland with hyperplasia and destruction,smaller CD1a⁺and S100⁺LC and its main distribution in hair follicle and sebaceous gland,obvious increase in LC'S quantity.The quantitative change of CD1a⁺and S100⁺LC was unobvious between at two?30min,72h?time point after moxibustion in TRPV1^-/-+38?moxi group?P>0.05?.Compared with theTRPV1^-/-+46?moxi group of of 30min after moxibustion,the number of CD1a⁺and S100⁺LC were significantly decreaseed in the one of 72h after moxibustion?P<0.05?.3.The multiple of quantitative change about langerhans cell:At 30min after moxibustion,the multiple of quantitative change about CD1a⁺langerhans cell in WT+46?moxi group?4.05±0.39?is notably higher than the one in TRPV1^-/-46?moxi group?1.32±0.28??P<0.05?;The multiple of quantitative change about S100⁺langerhans cell in WT+46?moxi group?1.16±0.06?is notably lower than the one in TRPV1^-/-46?moxi group?4.43±0.13??P<0.05?.At 72h after moxibustion,the multiple of quantitative change about CD1a⁺langerhans cell has no obvious difference between WT+46?moxi group?0.53±0.05?and TRPV1^-/-46?moxi group?0.38±0.06??P>0.05?;The multiple of quantitative change about S100⁺langerhans cell has no obvious difference between WT+46?moxi group?0.31±0.01?and TRPV1^-/-46?moxi group?0.25±0.03??P>0.05?;Conclusions:1.Moxibustion with different temperatures had different effect on the local skin structure of local acupoint in mice,and the influence of 46?moxibustion is greater than 38?moxibustion.2.Moxibustion with different temperatures had different influence on langerhans cell in acupoint.It indicates that immune response of local acupoint is relative to temperature.3.Knocking out the TRPV1 gene,the effect of 46?moxibustion on LC is not completely blocked and appears the same trend as the wide type mice.In addition,the multiple of quantitative change about langerhans cell has difference in different species.It indicates that moxibustion initiating skin immune function may be not completly depend on the TRPV1.