Observational studies have shown a negative correlation between Vitamin D level and the likelihood of developing insulin resistance (IR) and/or diabetes over time, yet evidence remains inconsistent. In this meta-analysis and systematic review, we strive to define the potential association between serum or supplemental Vitamin D Levels and insulin resistance respectively, as well as the contribution of Vitamin D to type 2 diabetes, and to summarize the biologic plausibility of Vitamin D. Four databases (PubMed, Embase, Cochrane Library, and Web of Science) were searched for this Systematic Literature Review (SLR) to find appropriate observational studies and clinical trials published in English through to July 2022. EndNote (version X9) is used to manage the literature search results. We calculated Standard Mean Differences (SMDs) and Risk Ratios (RRs) with their 95% Confidence Intervals (CIs), separately, for continuous and dichotomous outcomes. The correlation coefficients were normalized to z values through Fisher's z-transformation to calculate the relevant statistics. Meta-analyses were carried out for all comparisons, based on a random-effects pooling model. Data analysis was performed using RevMan (version 5.3) and STATA (version 15.1). All statistical tests were two-sided, with P < 0.05 were regarded as significant. In our current meta-analysis, there are 18 RCTs and 20 observational studies including 1243 and 11,063 participants respectively. In the overall analysis, the diabetic with Vitamin D supplement treatment group showed significantly improve serum insulin (SMD =  - 0.265, 95% CI - 0.394 to - 0.136, P < 0.05), glucose (SMD =  - 0.17, 95% CI - 0.301to - 0.039, P < 0.05) and HOMA-IR (SMD =  - 0.441, 95% CI - 0.582 to - 0.3, P < 0.05) compared with the routine treatment group. Correlation analysis results showed that all three outcomes were significantly correlated in a negative manner with raised Vitamin D (insulin: r =  - 0.08 95% =  - 0.12 to - 0.04; glucose: r =  - 0.06 95% =  - 0.11 to - 0.01; HOMA-IR: r =  - 0.08 95% =  - 0.09 to - 0.06). Results of overall analysis proved that vitamin D has shown significant effect on regulates insulin resistance, and there is a significant inverse association between serum Vitamin D level and IR. Vitamin D supplementation is expected to be integrated into conventional medical approaches to prevent type 2 diabetes and to mitigate the burden of diabetes for individuals and society.PROSPERO registration number: CRD42022348295.